Detection kit or device and detection method for biliary tract cancer

Abstract

The invention provides a kit or device for detecting biliary tract cancer, and a detection method. The present invention relates to: a kit or device for detecting biliary tract cancer, which contains a nucleic acid capable of specifically binding to miRNA in a specimen of a subject; and a method for detecting biliary tract cancer, which includes measuring the miRNA in vitro.

Description

[0001] This application is a divisional application of patent application filed on June 11, 2015, with application number 202010954168.1 and entitled "Detection kit or device for biliary tract cancer and detection method". Technical Field

[0002] This invention relates to a kit or apparatus for detecting biliary tract cancer, comprising a nucleic acid capable of specifically binding to a specific miRNA, for examining whether a subject has biliary tract cancer, and a method for detecting biliary tract cancer comprising using the nucleic acid to determine the expression level of the miRNA. Background Technology

[0003] The bile ducts refer to the main excretory pathway from the bile secreted by hepatocytes to its exit into the duodenum. They are broadly divided into the intrahepatic bile ducts within the liver and the extrahepatic bile duct system outside the liver. The extrahepatic bile duct system is roughly divided into three parts: the extrahepatic bile ducts that drain bile from the liver into the duodenum; the gallbladder, which temporarily stores and concentrates bile; and the duodenal papilla, the opening of the bile ducts and the main pancreatic duct into the lumen of the duodenum.

[0004] Most biliary tract cancers involve the malignant transformation of the epithelial cells lining the bile ducts. Chemotherapy and radiation therapy are less effective, and early detection and surgical resection are the only radical treatment. However, early-stage biliary tract cancers often present with no noticeable symptoms. For example, as the cancer progresses and the bile ducts become blocked, bile refluxes into the blood vessels, causing symptoms such as jaundice and itching. Therefore, these cancers are usually discovered in their advanced stages. Furthermore, intrahepatic cholangiocarcinomas rarely cause extrahepatic bile duct blockage, so most do not produce jaundice and progress asymptomatically. According to statistics on cancer mortality rates in Japan in 2011 published by the Cancer Countermeasures Information Center of the National Cancer Center, the number of deaths from biliary tract cancer rose to 18,186. The 5-year relative survival rate for each site from 2003 to 2005 was 22.5% for men and 19.9% ​​for women, second only to pancreatic cancer. The bile ducts are closely related to vital organs such as the liver and pancreas; therefore, metastasis to these organs is a major cause of poor prognosis.

[0005] Biliary duct cancer is broadly classified into three types based on its location: extrahepatic bile duct cancer, gallbladder cancer, and papillary bile duct cancer. Extrahepatic bile duct cancer is further divided into four types: hilar bile duct cancer (located at the entrance to the liver), upper bile duct cancer (extended bile duct cancer) (from the hilum to the upper part of the gallbladder), middle bile duct cancer (from the gallbladder to the middle part of the pancreas), and lower bile duct cancer (from the pancreas to the lower part of the duodenal papilla). It is known that the closer the bile duct is to the liver, the more difficult the surgery and the worse the prognosis.

[0006] The progression of extrahepatic bile duct carcinoma, gallbladder carcinoma, and papillary bile duct carcinoma based on the UICC (Unio Internationalis Contra Cancrum) is defined in "Biliary Tract Cancer Procedures, 5th Edition" (edited by the Japanese Society for Biliary Surgery, Kinbara Publishing Co., Ltd., 2003, p109), which classifies them into stages 0, IA, IB, IIA, IIB, III, IVa, and IVb based on lymph node metastasis, metastasis to other distant organs outside the abdominal cavity, and gross peribiliary progression. The progression of intrahepatic bile duct carcinoma based on the UICC is defined in "TNM Classification of Malignant Tumors, 7th Edition (Japanese Version)" (UICC Japan Committee, translated by the TNM Committee, Kinbara Publishing Co., Ltd., 2012, p110), which classifies it into stages I, II, III, IVa, and IVb based on lymph node metastasis, metastasis to other distant organs outside the abdominal cavity, and gross peribiliary progression.

[0007] The initial diagnosis of biliary tract cancer typically uses low-invasive blood biochemistry tests, tumor marker tests, and abdominal ultrasound (Non-Patent Literature 1). Blood biochemistry tests used to detect biliary tract cancer include, for example, alkaline phosphatase, γ-GTP, and bilirubin, which are elevated due to liver dysfunction. Known tumor markers for detecting biliary tract cancer include, for example, CEA, CA19-9, DUPAN-2, CA195, CA242, and IL-6. As a method of using these tumor markers, cancer is suspected when their blood concentration is higher or lower than a pre-set baseline value. For example, as described in Non-Patent Literature 2, a baseline value for CEA is set at 5 ng / mL, and a baseline value for CA19-9 is set at 37 U / mL; values ​​above these values ​​are suspected to indicate cancer containing biliary tract cancer.

[0008] In addition, although it is still in the research stage, there are reports of using the expression levels of proteins and genes in biological samples, such as blood, to detect biliary tract cancer.

[0009] Patent document 1 describes a method for detecting biliary tract cancer by using the expression levels of proteins in biliary tract tissue.

[0010] Patent document 2 describes a method for diagnosing gastrointestinal cancers, including biliary tract cancer, using mRNA genes extracted from cells (monocytes, etc.) in the blood.

[0011] Existing technical documents

[0012] Patent documents

[0013] Patent Document 1: Japanese Patent Application Publication No. 2012-237685

[0014] Patent Document 2: Japanese Patent Application Publication No. 2013-223520

[0015] Non-patent literature

[0016] Non-Patent Literature 1: Edited by the Committee for the Production and Publication of Guidelines for the Diagnosis and Treatment of Biliary Tract Cancer, "Evidence-Based Guidelines for the Diagnosis and Treatment of Biliary Tract Cancer", Medical Book Publishing Co., Ltd., 2007, pp. 38-39

[0017] Non-patent literature 2: Kiyoshi Kurokawa, Handbook of Clinical Examination Data, 2013, pp. 633, 636 Summary of the Invention

[0018] The problem that the invention aims to solve

[0019] The objective of this invention is to discover novel tumor markers for biliary tract cancer and to provide a method for effectively detecting biliary tract cancer using nucleic acids that can specifically bind to these markers. As described in Non-Patent Literature 1, the initial diagnosis of biliary tract cancer typically uses low-invasive blood biochemistry tests, tumor marker tests, and abdominal ultrasound. The tumor detection rate (the probability of cancer being detected by imaging) for biliary tract cancer using abdominal ultrasound ranges widely (21-90%), particularly decreasing when the cancer is located in the lower bile duct. Blood biochemistry tests, for example, for biliary tract cancer detection, also utilize indicators such as alkaline phosphatase, γ-GTP, and bilirubin, which are elevated due to liver dysfunction; however, these blood biochemistry tests do not specifically detect biliary tract cancer. Furthermore, known tumor markers for biliary tract cancer detection include, for example, CEA, CA19-9, DUPAN-2, CA195, CA242, and IL-6. Among these, CEA is known to rise in 40–70% of patients with biliary tract cancer, and CA19-9 is known to rise in 50–79% of patients with biliary tract cancer (Non-Patent Literature 1), but these markers are not specific to biliary tract cancer and are difficult to use for early diagnosis, as described in Non-Patent Literature 1. Furthermore, the clinical usefulness of DUPAN-2, CA195, CA242, and IL-6 is unclear, as described in Non-Patent Literature 1. Therefore, it is also considered that the use of conventional tumor markers may lead to misdiagnosis as other cancers, and / or benign tumors and / or benign diseases of the bile duct and / or surrounding organs.

[0020] Furthermore, although it is still in the research stage, there are reports such as using the expression levels of proteins and genes in biological samples, such as blood, to detect biliary tract cancer, but none of them have been put into practical use.

[0021] Patent Document 1 describes a method for detecting biliary tract cancer using the expression levels of proteins in bile duct tissue. However, this method requires surgical removal of the tissue to obtain a sample, which places a heavy physical burden on the patient, making it an undesirable method. Furthermore, Patent Document 1 does not describe the specific accuracy, sensitivity, specificity, or other performance characteristics of this detection method for identifying biliary tract cancer, thus lacking industrial applicability.

[0022] Patent Document 2 describes a method for diagnosing gastrointestinal cancers, including biliary tract cancer, using mRNA genes extracted from blood cells (monocytes, etc.). However, this detection method requires the combined use of dozens to hundreds of mRNAs, raising concerns about increased testing costs and the complexity of the discrimination algorithm in the development of a practical test. Furthermore, mRNA is easily degraded and unstable in blood, making it less than ideal as a testing subject.

[0023] Thus, in the detection of biliary tract cancer, existing tumor markers have low performance, or for markers in the research stage, the performance and detection methods are not specifically described. Therefore, using them carries the following risks: unnecessary additional testing may be performed due to misdiagnosing healthy individuals as biliary tract cancer patients, or treatment opportunities may be lost due to overlooking biliary tract cancer patients. Furthermore, measuring dozens to hundreds of genes increases the cost of testing, making it difficult to use for large-scale screening such as health diagnostics. Additionally, collecting biliary tract tissue for tumor marker measurement is highly invasive and not preferable for patients. Therefore, there is a need for biliary tract cancer markers that can be detected from blood with low invasiveness, accurately distinguish biliary tract cancer patients from biliary tract cancer patients, and accurately distinguish healthy individuals from healthy individuals. In particular, since early detection and resection are the only radical treatment for biliary tract cancer, there is an urgent need for highly sensitive biliary tract cancer markers.

[0024] Methods for solving problems

[0025] In order to solve the above-mentioned problems, the inventors conducted in-depth research and found multiple genes that can be used as biomarkers for the detection of biliary tract cancer from blood that can be collected in a low-invasive manner. They also found that biliary tract cancer can be significantly detected by using nucleic acids that can specifically bind to these genes, thus completing the present invention.

[0026] <Summary of the Invention>

[0027] That is, the present invention has the following features.

[0028] (1) A kit for detecting biliary tract cancer, which comprises agents capable of binding to miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6836-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4651, miR-8069, miR-1238-5p, miR-6880-5p, miR-8072, miR-4723-5p, miR-4732-5p, miR-6125, miR-6090, miR-7114-5p, miR-564, miR-451a, miR-3135b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3p, miR-1227-5p, miR-3188, miR-7704, miR-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a, miR-4707-5p, miR-6741-5p, miR-1260b, miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-4433b-3p, miR-197-5p, miR-718, miR-4513, miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449, which are used as biliary tract cancer markers.miR-6889-5p,miR-486-3p,miR-7113-3p,miR-642a-3p,miR-7847-3p,miR-6768-5p,miR-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR -4467,miR-6858-5p,miR-4417,miR-3665,miR-4736,miR-4687-3p,miR-1908-5p,miR-5195-3p,miR-4286,miR-3679-3p,miR-6791-5p,miR-1202,miR-3 Nucleic acids specifically bound to at least one of the following polynucleotides: miR-4746-3p, miR-3184-5p, miR-3937, miR-6515-3p, miR-6132, miR-187-5p, miR-7111-5p, miR-5787, miR-6779-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p.

[0029] (2) According to the kit described in (1), wherein miR-125a-3p is hsa-miR-125a-3p, miR-6893-5p is hsa-miR-6893-5p, miR-204-3p is hsa-miR-204-3p, miR-4476 is hsa-miR-4476, miR-4294 is hsa-miR-4294, miR-150-3p is hsa-miR-150-3p, miR-6729-5p is hsa-miR-6729-5p, miR-7641 is hsa-miR-7641, miR-6765-3p is hsa-miR-6765-3p, miR -6820-5p is hsa-miR-6820-5p, miR-575 is hsa-miR-575, miR-6836-3p is hsa-miR-6836-3p, miR-1469 is hsa-miR-1469, miR-663a is hsa-miR-663a, miR-6075 is hsa-miR-6075, miR-4634 is hsa-miR-4634, miR-423-5p is hsa-miR-423-5p, miR-4454 is hsa-miR-4454, miR-7109-5p is hsa-miR-7109-5p, miR-6789-5p is h sa-miR-6789-5p, miR-6877-5p is hsa-miR-6877-5p, miR-4792 is hsa-miR-4792, miR-4530 is hsa-miR-4530, miR-7975 is hsa-miR-7975, miR-6724-5p is hsa-miR-6724-5p, miR-8073 is hsa-miR-8073, miR-7977 is hsa-miR-7977, miR-1231 is hsa-miR-1231, miR-6799-5p is hsa-miR-6799-5p, miR-615-5p is hsa-miR- 615-5p, miR-4450 is hsa-miR-4450, miR-6726-5p is hsa-miR-6726-5p, miR-6875-5p is hsa-miR-6875-5p, miR-4734 is hsa-miR-4734, miR-16-5p is hsa-miR-16-5p, miR-602 is hsa-miR-602, miR-4651 is hsa-miR-4651, miR-8069 is hsa-miR-8069, miR-1238-5p is hsa-miR-1238-5p, miR-6880-5p is hsa-miR-6880-5p.miR-8072 is hsa-miR-8072, miR-4723-5p is hsa-miR-4723-5p, miR-4732-5p is hsa-miR-4732-5p, miR-6125 is hsa-miR-6125, miR-6090 is hsa-miR-6090, miR-7114-5p is hsa-miR-7114-5p, miR-564 is hsa-miR-564, miR-451a is hsa-miR-451a, miR-3135b is hsa-miR-3135b, miR-4497 is hsa-miR-4497, miR-466 5-5p is hsa-miR-4665-5p, miR-3622a-5p is hsa-miR-3622a-5p, miR-6850-5p is hsa-miR-6850-5p, miR-6821-5p is hsa-miR-6821-5p, miR-5100 is hsa-miR-5100, miR-6872-3p is hsa-miR-6872-3p, miR-4433-3p is hsa-miR-4433-3p, miR-1227-5p is hsa-miR-1227-5p, miR-3188 is hsa-miR-3188, miR-7704 is hs a-miR-7704, miR-3185 is hsa-miR-3185, miR-1908-3p is hsa-miR-1908-3p, miR-6781-5p is hsa-miR-6781-5p, miR-6805-5p is hsa-miR-6805-5p, miR-8089 is hsa-miR-8089, miR-665 is hsa-miR-665, miR-4486 is hsa-miR-4486, miR-6722-3p is hsa-miR-6722-3p, miR-1260a is hsa-miR-1260a, miR-4707-5p is... hsa-miR-4707-5p, miR-6741-5p is hsa-miR-6741-5p, miR-1260b is hsa-miR-1260b, miR-1246 is hsa-miR-1246, miR-6845-5p is hsa-miR-6845-5p, miR-4638-5p is hsa-miR-4638-5p, miR-6085 is hsa-miR-6085, miR-1228-3p is hsa-miR-1228-3p, miR-4534 is hsa-miR-4534, miR-5585-3p is hsa-miR-5585-3p.miR-4741 is hsa-miR-4741, miR-4433b-3p is hsa-miR-4433b-3p, miR-197-5p is hsa-miR-197-5p, miR-718 is hsa-miR-718, miR-4513 is hsa-miR-4513, miR-4446-3p is hsa-miR-4446-3p, miR-619-5p is hsa-miR-619-5p, miR-6816-5p is hsa-miR-6816-5p, miR-6778-5p is hsa-miR-6778-5p, miR-24-3p is hsa -miR-24-3p, miR-1915-3p is hsa-miR-1915-3p, miR-4665-3p is hsa-miR-4665-3p, miR-4449 is hsa-miR-4449, miR-6889-5p is hsa-miR-6889-5p, miR-486-3p is hsa-miR-486-3p, miR-7113-3p is hsa-miR-7113-3p, miR-642a-3p is hsa-miR-642a-3p, miR-7847-3p is hsa-miR-7847-3p, miR-6768-5p is hsa-m iR-6768-5p, miR-1290 is hsa-miR-1290, miR-7108-5p is hsa-miR-7108-5p, miR-92b-5p is hsa-miR-92b-5p, miR-663b is hsa-miR-663b, miR-3940-5p is hsa-miR-3940-5p, miR-4467 is hsa-miR-4467, miR-6858-5p is hsa-miR-6858-5p, miR-4417 is hsa-miR-4417, miR-3665 is hsa-miR-3665, miR-4736 is hsa -miR-4736, miR-4687-3p is hsa-miR-4687-3p, miR-1908-5p is hsa-miR-1908-5p, miR-5195-3p is hsa-miR-5195-3p, miR-4286 is hsa-miR-4286, miR-3679-3p is hsa-miR-3679-3p, miR-6791-5p is hsa-miR-6791-5p, miR-1202 is hsa-miR-1202, miR-3656 is hsa-miR-3656, miR-4746-3p is hsa-miR-4746-3p.miR-3184-5p is hsa-miR-3184-5p, miR-3937 is hsa-miR-3937, miR-6515-3p is hsa-miR-6515-3p, miR-6132 is hsa-miR-6132, miR-187-5p is hsa-miR-187-5p, miR-7111-5p is hsa-miR-7111-5p, miR-5787 is hsa-miR-5787, and miR-6779-5p is hsa-miR-6779-5p, miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4649-5p, miR-760 The values ​​are hsa-miR-760, hsa-miR-3162-5p, hsa-miR-3178, hsa-miR-3178, hsa-miR-940, hsa-miR-940, hsa-miR-4271, hsa-miR-4271, hsa-miR-6769b-5p, hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-6757-5p, hsa-miR-3131, and hsa-miR-1343-3p.

[0030] (3) According to the kit described in (1) or (2), the above nucleic acid is a polynucleotide selected from the polynucleotides shown in (a) to (e) below.

[0031] (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0032] (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 1–125 and 466–478.

[0033] (c) A polynucleotide, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases, consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t.

[0034] (d) A polynucleotide containing a base sequence complementary to the base sequence shown in any of the sequence numbers 1–125, 466–478, or a base sequence in which u is t, and

[0035] (e) A polynucleotide that hybridizes with any of the polynucleotides (a) to (d) above under strict conditions.

[0036] (4) The kit according to any one of (1) to (3), wherein the kit further comprises a reagent capable of reacting with other biliary tract cancer markers selected from miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, m Nucleic acids specifically bound to at least one of the following polynucleotides: iR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, and miR-6780b-5p.

[0037] (5) The kit according to (4), wherein miR-6808-5p is hsa-miR-6808-5p, miR-6774-5p is hsa-miR-6774-5p, miR-4656 is hsa-miR-4656, miR-6806-5p is hsa-miR-6806-5p, and miR-1233-5p is hsa-miR-1233-5p. miR-328-5p is hsa-miR-328-5p, miR-4674 is hsa-miR-4674, miR-2110 is hsa-miR-2110, miR-6076 is hsa-miR-6076, miR-3619-3p is hsa-miR-3619-3p, and miR-92a-2-5p is hsa-miR-92a-2-5p. miR-128-1-5p is hsa-miR-128-1-5p, miR-638 is hsa-miR-638, miR-2861 is hsa-miR-2861, miR-371a-5p is hsa-miR-371a-5p, miR-211-3p is hsa-miR-211-3p, and miR-1273g-3p is hsa-miR-12 73g-3p, miR-1203 is hsa-miR-1203, miR-122-5p is hsa-miR-122-5p, miR-4258 is hsa-miR-4258, miR-4484 is hsa-miR-4484, miR-4648 is hsa-miR-4648, and miR-6780b-5p is hsa-miR-6780b-5p.

[0038] (6) According to the kit described in (4) or (5), the above nucleic acid is a polynucleotide selected from the polynucleotides shown in (f) to (j) below.

[0039] (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0040] (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126–148.

[0041] (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequences 126 to 148 or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0042] (i) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequences 126–148 or a base sequence in which u is t, and

[0043] (j) A polynucleotide that hybridizes with any of the polynucleotides (f) to (i) above under strict conditions.

[0044] (7) The kit according to any one of (1) to (6), wherein the kit contains at least two nucleic acids capable of specifically binding to at least two polynucleotides selected from all biliary tract cancer markers described in (1) or (2).

[0045] (8). 3p、miR-4476、miR-4294、miR-150-3p、miR-6729-5p、miR-7641、miR-6765- 3p, miR-6820-5p, miR-575, miR-6836-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877- 5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p miR-4734 miR-16-5p miR-602 miR-4651 miR-8069 miR-1238-5p miR-6880-5p miR-8072 miR-4723-5p miR-4732-5p miR-6125 miR-6090 miR- 7114-5p, miR-564, miR-451a, miR-3135b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3 p, miR-1227-5p, miR-3188, miR-7704, miR-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a miR-4707-5p, miR-6741-5p, miR-1260b, miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-44 33b-3p, miR-197-5p, miR-718, miR-4513, miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449miR-6889-5p,miR-486-3p,miR-7113-3p,miR-642a-3p,miR-7847-3p,miR-6768-5p,miR-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR -4467,miR-6858-5p,miR-4417,miR-3665,miR-4736,miR-4687-3p,miR-1908-5p,miR-5195-3p,miR-4286,miR-3679-3p,miR-6791-5p,miR-1202,miR-3 Nucleic acids specifically bound to at least one of the following polynucleotides: miR-4746-3p, miR-3184-5p, miR-3937, miR-6515-3p, miR-6132, miR-187-5p, miR-7111-5p, miR-5787, miR-6779-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p.

[0046] (9) The apparatus according to (8), wherein miR-125a-3p is hsa-miR-125a-3p, miR-6893-5p is hsa-miR-6893-5p, miR-204-3p is hsa-miR-204-3p, miR-4476 is hsa-miR-4476, miR-4294 is hsa-miR-4294, miR-150-3p is hsa-miR-150-3p, miR-6729-5p is hsa-miR-6729-5p, miR-7641 is hsa-miR-7641, miR-6765-3p is hsa-miR-6765-3p, miR- 6820-5p is hsa-miR-6820-5p, miR-575 is hsa-miR-575, miR-6836-3p is hsa-miR-6836-3p, miR-1469 is hsa-miR-1469, miR-663a is hsa-miR-663a, miR-6075 is hsa-miR-6075, miR-4634 is hsa-miR-4634, miR-423-5p is hsa-miR-423-5p, miR-4454 is hsa-miR-4454, miR-7109-5p is hsa-miR-7109-5p, miR-6789-5p is hs a-miR-6789-5p, miR-6877-5p is hsa-miR-6877-5p, miR-4792 is hsa-miR-4792, miR-4530 is hsa-miR-4530, miR-7975 is hsa-miR-7975, miR-6724-5p is hsa-miR-6724-5p, miR-8073 is hsa-miR-8073, miR-7977 is hsa-miR-7977, miR-1231 is hsa-miR-1231, miR-6799-5p is hsa-miR-6799-5p, miR-615-5p is hsa-miR-6 15-5p, miR-4450 is hsa-miR-4450, miR-6726-5p is hsa-miR-6726-5p, miR-6875-5p is hsa-miR-6875-5p, miR-4734 is hsa-miR-4734, miR-16-5p is hsa-miR-16-5p, miR-602 is hsa-miR-602, miR-4651 is hsa-miR-4651, miR-8069 is hsa-miR-8069, miR-1238-5p is hsa-miR-1238-5p, miR-6880-5p is hsa-miR-6880-5p.miR-8072 is hsa-miR-8072, miR-4723-5p is hsa-miR-4723-5p, miR-4732-5p is hsa-miR-4732-5p, miR-6125 is hsa-miR-6125, miR-6090 is hsa-miR-6090, miR-7114-5p is hsa-miR-7114-5p, miR-564 is hsa-miR-564, miR-451a is hsa-miR-451a, miR-3135b is hsa-miR-3135b, miR-4497 is hsa-miR-4497, miR-466 5-5p is hsa-miR-4665-5p, miR-3622a-5p is hsa-miR-3622a-5p, miR-6850-5p is hsa-miR-6850-5p, miR-6821-5p is hsa-miR-6821-5p, miR-5100 is hsa-miR-5100, miR-6872-3p is hsa-miR-6872-3p, miR-4433-3p is hsa-miR-4433-3p, miR-1227-5p is hsa-miR-1227-5p, miR-3188 is hsa-miR-3188, miR-7704 is hs a-miR-7704, miR-3185 is hsa-miR-3185, miR-1908-3p is hsa-miR-1908-3p, miR-6781-5p is hsa-miR-6781-5p, miR-6805-5p is hsa-miR-6805-5p, miR-8089 is hsa-miR-8089, miR-665 is hsa-miR-665, miR-4486 is hsa-miR-4486, miR-6722-3p is hsa-miR-6722-3p, miR-1260a is hsa-miR-1260a, miR-4707-5p is... hsa-miR-4707-5p, miR-6741-5p is hsa-miR-6741-5p, miR-1260b is hsa-miR-1260b, miR-1246 is hsa-miR-1246, miR-6845-5p is hsa-miR-6845-5p, miR-4638-5p is hsa-miR-4638-5p, miR-6085 is hsa-miR-6085, miR-1228-3p is hsa-miR-1228-3p, miR-4534 is hsa-miR-4534, miR-5585-3p is hsa-miR-5585-3p.miR-4741 is hsa-miR-4741, miR-4433b-3p is hsa-miR-4433b-3p, miR-197-5p is hsa-miR-197-5p, miR-718 is hsa-miR-718, miR-4513 is hsa-miR-4513, miR-4446-3p is hsa-miR-4446-3p, miR-619-5p is hsa-miR-619-5p, miR-6816-5p is hsa-miR-6816-5p, miR-6778-5p is hsa-miR-6778-5p, miR-24-3p is hsa -miR-24-3p, miR-1915-3p is hsa-miR-1915-3p, miR-4665-3p is hsa-miR-4665-3p, miR-4449 is hsa-miR-4449, miR-6889-5p is hsa-miR-6889-5p, miR-486-3p is hsa-miR-486-3p, miR-7113-3p is hsa-miR-7113-3p, miR-642a-3p is hsa-miR-642a-3p, miR-7847-3p is hsa-miR-7847-3p, miR-6768-5p is hsa-m iR-6768-5p, miR-1290 is hsa-miR-1290, miR-7108-5p is hsa-miR-7108-5p, miR-92b-5p is hsa-miR-92b-5p, miR-663b is hsa-miR-663b, miR-3940-5p is hsa-miR-3940-5p, miR-4467 is hsa-miR-4467, miR-6858-5p is hsa-miR-6858-5p, miR-4417 is hsa-miR-4417, miR-3665 is hsa-miR-3665, miR-4736 is hsa -miR-4736, miR-4687-3p is hsa-miR-4687-3p, miR-1908-5p is hsa-miR-1908-5p, miR-5195-3p is hsa-miR-5195-3p, miR-4286 is hsa-miR-4286, miR-3679-3p is hsa-miR-3679-3p, miR-6791-5p is hsa-miR-6791-5p, miR-1202 is hsa-miR-1202, miR-3656 is hsa-miR-3656, miR-4746-3p is hsa-miR-4746-3p.miR-3184-5p is hsa-miR-3184-5p, miR-3937 is hsa-miR-3937, miR-6515-3p is hsa-miR-6515-3p, miR-6132 is hsa-miR-6132, miR-187-5p is hsa-miR-187-5p, miR-7111-5p is hsa-miR-7111-5p, miR-5787 is hsa-miR-5787, and miR-6779-5p is hsa-miR-6779-5p, miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4649-5p, miR-760 The values ​​are hsa-miR-760, hsa-miR-3162-5p, hsa-miR-3178, hsa-miR-3178, hsa-miR-940, hsa-miR-940, hsa-miR-4271, hsa-miR-4271, hsa-miR-6769b-5p, hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-6757-5p, hsa-miR-3131, and hsa-miR-1343-3p.

[0047] (10) In the apparatus according to (8) or (9), the nucleic acid is a polynucleotide selected from the polynucleotides shown in (a) to (e) below.

[0048] (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0049] (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 1–125 and 466–478.

[0050] (c) A polynucleotide, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases, consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t.

[0051] (d) A polynucleotide containing a base sequence complementary to the base sequence shown in any of the sequence numbers 1–125, 466–478, or a base sequence in which u is t, and

[0052] (e) A polynucleotide that hybridizes with any of the polynucleotides (a) to (d) above under strict conditions.

[0053] (11) The device according to any one of (8) to (10), the device further comprising the ability to interact with other biliary tract cancer markers selected from miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, m Nucleic acids specifically bound to at least one of the following polynucleotides: iR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, and miR-6780b-5p.

[0054] (12) The apparatus according to (11), wherein miR-6808-5p is hsa-miR-6808-5p, miR-6774-5p is hsa-miR-6774-5p, miR-4656 is hsa-miR-4656, miR-6806-5p is hsa-miR-6806-5p, and miR-1233-5p is hsa-miR-1233-5p. p, miR-328-5p is hsa-miR-328-5p, miR-4674 is hsa-miR-4674, miR-2110 is hsa-miR-2110, miR-6076 is hsa-miR-6076, miR-3619-3p is hsa-miR-3619-3p, miR-92a-2-5p is hsa-miR-92a-2-5p miR-128-1-5p is hsa-miR-128-1-5p, miR-638 is hsa-miR-638, miR-2861 is hsa-miR-2861, miR-371a-5p is hsa-miR-371a-5p, miR-211-3p is hsa-miR-211-3p, and miR-1273g-3p is hsa-miR-12 73g-3p, miR-1203 is hsa-miR-1203, miR-122-5p is hsa-miR-122-5p, miR-4258 is hsa-miR-4258, miR-4484 is hsa-miR-4484, miR-4648 is hsa-miR-4648, and miR-6780b-5p is hsa-miR-6780b-5p.

[0055] (13) In the apparatus according to (11) or (12), the nucleic acid is a polynucleotide selected from the polynucleotides shown in (f) to (j) below.

[0056] (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0057] (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126–148.

[0058] (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequences 126 to 148 or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0059] (i) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequences 126–148 or a base sequence in which u is t, and

[0060] (j) A polynucleotide that hybridizes with any of the polynucleotides (f) to (i) above under strict conditions.

[0061] (14) The apparatus according to any one of (8) to (13) is an apparatus for determination by hybridization technique.

[0062] (15) According to the apparatus described in (14), the hybridization technique is a nucleic acid array technique.

[0063] (16) The device according to any one of (8) to (15) comprises at least two nucleic acids capable of specifically binding to at least two polynucleotides selected from all biliary cancer markers described in (8) or (9).

[0064] (17) A method for detecting biliary tract cancer, comprising: using a kit described in any one of (1) to (7) or an apparatus described in any one of (8) to (16), measuring the expression level of a target nucleic acid in a sample of a test subject, and using the measured expression level and the control expression level of a healthy body measured similarly to evaluate in vitro whether the test subject has biliary tract cancer or not.

[0065] (18) According to the method described in (17), the subject is a human.

[0066] (19) The specimen is blood, serum or plasma, as described in (17) or (18).

[0067] <Definitions of Terms>

[0068] The terms used in this specification have the following definitions.

[0069] In this specification, "biliary tract cancer" refers to all malignant tumors that form in the bile ducts. Specifically, it includes extrahepatic bile duct cancer, gallbladder cancer, papillary bile duct cancer, duodenal papillary bile duct cancer, and intrahepatic bile duct cancer.

[0070] In this specification, "benign tumors and / or benign diseases of the bile duct and / or surrounding organs" refers to non-malignant tumors related to the bile duct, liver, and pancreas.

[0071] The abbreviations for nucleotides, polynucleotides, DNA, RNA, etc., shall be in accordance with the “Guidelines for the Production of Instructions, etc., Containing Base Sequences or Amino Acid Sequences” (edited by the Japanese Patent Office) and common usage in this technical field.

[0072] In this specification, "polynucleotide" refers to nucleic acids comprising any of RNA, DNA, and RNA / DNA (chimeras). Furthermore, the aforementioned DNA includes any of cDNA, genomic DNA, and synthetic DNA. Additionally, the aforementioned RNA includes any of total RNA, mRNA, rRNA, miRNA, siRNA, snoRNA, snRNA, non-coding RNA, and synthetic RNA. In this specification, "synthetic DNA" and "synthetic RNA" refer to DNA and RNA artificially produced based on specified base sequences (which can be either native or non-native sequences), using, for example, an automated nucleic acid synthesizer. In this specification, "non-native sequence" is intended to be used in a broad sense, encompassing sequences that differ from native sequences, such as sequences containing one or more substitutions, deletions, insertions, and / or additions (i.e., mutant sequences), sequences containing one or more modified nucleotides (i.e., modified sequences), etc. Furthermore, in this specification, polynucleotides and nucleic acids are used interchangeably.

[0073] In this specification, a “fragment” is a polynucleotide having a continuous portion of a polynucleotide sequence, preferably having a length of 15 or more bases, more preferably 17 or more bases, and more preferably 19 or more bases.

[0074] In this specification, the term "gene" is intended to include not only RNA and double-stranded DNA, but also the individual single-stranded DNA strands that constitute them, such as the positive (or sense) strand or the complementary (or antisense) strand. Furthermore, there is no particular limitation on its length.

[0075] Therefore, unless otherwise specified, the term "gene" in this specification includes: double-stranded DNA containing human genomic DNA, single-stranded DNA (positive strand), single-stranded DNA containing cDNA with a sequence complementary to the positive strand (complementary strand), microRNA (miRNA), fragments thereof, and transcripts thereof. Furthermore, "gene" is not only the "gene" indicated by a specific base sequence (or sequence number), but also includes RNA with biological functions equivalent to the RNA encoded by them, such as "nucleic acids" encoding homologs (i.e., orthologs), gene polymorphisms, and derivatives. Specifically, examples of such "nucleic acids" encoding homologs, mutants, or derivatives include those having a base sequence that hybridizes to the complementary sequence of any of the base sequences indicated by sequence numbers 1 to 509 or a base sequence in which u is t, under the strict conditions described below. Additionally, "gene" may include, for example, expression control regions, coding regions, exons, or introns, regardless of differences in functional regions. Furthermore, "genes" can be contained within cells, released outside cells and exist independently, or be enclosed in vesicles called exogenous bodies.

[0076] In this specification, "exosome" refers to a small vesicle surrounded by a lipid bilayer secreted by cells. Exosomes originate from multivesicular endosomes and, when released into the extracellular environment, sometimes contain biological substances such as RNA, DNA (genes), and proteins. Exosomes are known to be found in bodily fluids such as blood, serum, plasma, and lymph.

[0077] In this specification, "transcription product" refers to RNA synthesized using a gene's DNA sequence as a template. RNA polymerase binds to a site upstream of the gene called the promoter, synthesizing RNA by binding ribonucleotides to the 3' end in a manner complementary to the DNA's base sequence. This RNA contains not only the gene itself but also the complete sequence from the transcription start site to the end of the poly-A sequence, represented by expression control regions, coding regions, exons, or introns.

[0078] Furthermore, unless otherwise specified, “microRNA (miRNA)” in this specification is intended to be used as a 15-25 base non-coding RNA that is transcribed from a hairpin-like RNA precursor, cleaved by a dsRNA cleaving enzyme with RNase III activity, encapsulated in a protein complex called RISC, and involved in the translational repression of mRNA. In addition, the term “miRNA” as used in this specification includes not only the “miRNA” indicated by the specific base sequence (or sequence number), but also its precursors (pre-miRNA, pri-miRNA), and miRNAs with equivalent biological functions, such as homologs (i.e., homologs or orthologs), mutants of gene polymorphisms, and derivatives. Specifically, examples of such precursors, homologs, mutants, or derivatives include “miRNAs” that can be identified by miRBase version 20 (http: / / www.mirbase.org / ) and have a base sequence that hybridizes to the complementary sequence of any specific base sequence indicated by any of the sequence numbers 1-509 under the stringent conditions described below. In addition, the term “miRNA” used in this specification may refer to the gene product of a miR gene, which contains mature miRNA (e.g., a 15-25 base or 19-25 base non-coding RNA involved in the translation repression of mRNA as described above) or miRNA precursor (e.g., pre-miRNA or pri-miRNA as described above).

[0079] In this specification, "probe" includes polynucleotides and / or complementary polynucleotides used to specifically detect RNA or polynucleotides derived from gene expression.

[0080] In this specification, "primers" refers to polynucleotides and / or complementary polynucleotides that specifically identify and amplify RNA produced through gene expression or polynucleotides derived from such RNA.

[0081] Here, complementary polynucleotides (complementary strands, negative strands) refer to polynucleotides that are complementary in base pairing based on a base pairing such as A:T(U) or G:C, consisting of a full-length sequence or a portion thereof (here referred to as the positive strand for convenience) of a polynucleotide composed of a base sequence defined by any of the sequence numbers 1 to 509, or a sequence of bases in that sequence where u is t. However, such complementary strands are not limited to the case where the base sequence of the target positive strand is completely complementary; they can also have a complementary relationship to the extent that they can hybridize with the target positive strand under strict conditions.

[0082] In this specification, "stringent conditions" refer to conditions under which the nucleic acid probe hybridizes to its target sequence to a greater extent than that for other sequences (e.g., a value greater than the average background measurement plus the standard error of the background measurement multiplied by 2). Stringent conditions are sequence-dependent and vary depending on the environment in which hybridization takes place. By controlling the stringency of hybridization and / or washing conditions, target sequences that are 100% complementary to the nucleic acid probe can be identified. Specific examples of "stringent conditions" are described later.

[0083] In this specification, "Tm value" refers to the temperature at which the double-stranded portion of a polynucleotide denatures into a single strand, with the double strand and single strand existing in a 1:1 ratio.

[0084] In this specification, "mutant" in the case of nucleic acids refers to a naturally occurring mutant caused by polymorphism, mutation, etc., or a mutant containing one or more deletions, substitutions, additions, or insertions of one or more bases in any of the base sequences from sequence number 1 to 509, or in the base sequences where u is t, or in a portion thereof; or a mutant showing approximately 90%, 95%, 97%, 98%, or 99% or more % identity with each of the base sequences or a portion thereof; or a nucleic acid that hybridizes with a polynucleotide or oligonucleotide containing the base sequence or a portion thereof under the strict conditions defined above.

[0085] In this specification, "multiple" refers to an integer of approximately 10, 9, 8, 7, 6, 5, 4, 3, or 2.

[0086] The mutants described in this manual can be created using known techniques such as site-directed mutagenesis and mutation introduction using PCR.

[0087] The "% identity" in this specification can be determined using the protein or gene search systems described above, with or without vacancies (Zheng Zhang et al., 2000, J. Comput. Biol., Vol. 7, pp. 203-214; Altschul, SF et al., 1990, Journal of Molecular Biology, Vol. 215, pp. 403-410; Pearson, WR et al., 1988, Proc. Natl. Acad. Sci. USA, Vol. 85, pp. 2444-2448).

[0088] In this specification, "derivative" refers to modified nucleic acids, and does not specifically include, for example, labeled derivatives using fluorophores, derivatives containing modified nucleotides (e.g., nucleotides containing halogen, alkyl such as methyl, alkoxy such as methoxy, thio, carboxymethyl, etc., and nucleotides subjected to base reconstruction, double bond saturation, deamination, substitution of oxygen molecules for sulfur molecules, etc.), PNA (peptide nucleic acid; Nielsen, PE et al., 1991, Science, Vol. 254, pp. 1497-500), LNA (locked nucleic acid; Obika, S. et al., 1998, Tetrahedron Lett., Vol. 39, pp. 5401-5404), etc.

[0089] The "nucleic acid" in this specification, capable of specifically binding to polynucleotides selected from the above-mentioned miRNA group used as markers of biliary tract cancer, is a synthetic or modulated nucleic acid. Specifically, it includes "nucleic acid probes" or "primers" and is used directly or indirectly to detect the presence of biliary tract cancer in a subject, or to diagnose the presence, severity, improvement, and sensitivity to biliary tract cancer treatment, or to screen candidate substances useful for the prevention, improvement, or treatment of biliary tract cancer. These include nucleotides, oligonucleotides, and polynucleotides that can specifically identify and bind to the transcripts or cDNA synthesized from any of the sequences 1-509 in organisms, particularly in bodily fluids such as blood and urine, associated with the development of biliary tract cancer. Based on the aforementioned properties, these nucleotides, oligonucleotides, and polynucleotides can be effectively used as probes for detecting the aforementioned genes expressed in organisms, tissues, and cells, and also as primers for amplifying the aforementioned genes expressed in organisms.

[0090] The term "test" as used in this manual may be replaced with terms such as examination, measurement, detection, or judgment support. Furthermore, the term "evaluation" in this manual is used to imply support for diagnosis or evaluation based on examination or measurement results.

[0091] The term "test subject" as used in this instruction manual refers to mammals including humans, chimpanzees, pet animals such as dogs and cats, livestock such as cattle, horses, sheep, and goats, and rodents such as mice and rats. Furthermore, "healthy subject" also refers to such mammals, meaning animals that do not have the cancer being tested for.

[0092] As used in this specification, "P" or "P-value" indicates, in a statistical test, the probability that an extreme statistic will be observed compared to the actual statistic calculated from the data, under the null hypothesis. Therefore, the smaller the "P" or "P-value," the more significant the difference is considered between the compared objects.

[0093] In this instruction manual, "sensitivity" refers to the value of (number of true positives) / (number of true positives + number of false negatives). High sensitivity allows for early detection of biliary tract cancer, leading to complete resection of the cancerous portion and a reduced recurrence rate.

[0094] In this instruction manual, "specificity" refers to (number of true negatives) / (number of true negatives + number of false positives). High specificity prevents unnecessary additional testing due to misdiagnosing healthy individuals as having biliary tract cancer, thus reducing the burden on patients and lowering medical costs.

[0095] In this instruction manual, "accuracy" refers to the value of (number of true positives + number of true negatives) / (total number of cases). Accuracy represents the proportion of correct judgments for all samples and is the primary indicator for evaluating test performance.

[0096] In this specification, the term "specimen" as the object of judgment, detection, or diagnosis refers to tissues and biological materials whose gene expression changes as bile duct cancer develops, progresses, and is effectively treated. Specifically, it refers to bile duct tissue and its surrounding blood vessels, lymph nodes, and organs. It also includes organs suspected of metastasis, skin, and bodily fluids such as blood, urine, saliva, sweat, and tissue exudate, as well as serum, plasma, feces, and hair. Further, it refers to biological samples extracted from these, specifically genes such as RNA and miRNA.

[0097] The terms "hsa-miR-125a-3p gene" or "hsa-miR-125a-3p" as used in this specification include the hsa-miR-125a-3p gene described in sequence number 1 (miRBase accession number MIMAT0004602), homologs or orthologs of other species, etc. The hsa-miR-125a-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, CurrBiol, Vol. 12, pp. 735-739. In addition, as a precursor of "hsa-miR-125a-3p", "hsa-mir-125a" (miRBase accession number MI0000469, sequence number 149), which forms a hairpin-like structure, is known.

[0098] The terms "hsa-miR-6893-5p gene" or "hsa-miR-6893-5p" as used in this specification include the hsa-miR-6893-5p gene described in sequence number 2 (miRBase accession number MIMAT0027686), homologs or orthologs of other species, etc. The hsa-miR-6893-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6893-5p", "hsa-mir-6893" (miRBase accession number MI0022740, sequence number 150) is known to form a hairpin-like structure.

[0099] The terms "hsa-miR-204-3p gene" or "hsa-miR-204-3p" as used in this specification include the hsa-miR-204-3p gene described in sequence number 3 (miRBase accession number MIMAT0022693), homologs or orthologs of other species, etc. The hsa-miR-204-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. In addition, as a precursor of "hsa-miR-204-3p", "hsa-mir-204" (miRBase accession number MI0000284, sequence number 151) is known to form a hairpin-like structure.

[0100] The terms "hsa-miR-4476 gene" or "hsa-miR-4476" as used in this specification include the hsa-miR-4476 gene described in sequence number 4 (miRBase accession number MIMAT0019003), homologs or orthologs of other species, etc. The hsa-miR-4476 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4476", "hsa-mir-4476" (miRBase accession number MI0016828, sequence number 152) is known to form a hairpin-like structure.

[0101] The terms "hsa-miR-4294 gene" or "hsa-miR-4294" as used in this specification include the hsa-miR-4294 gene described in sequence number 5 (miRBase accession number MIMAT0016849), homologs or orthologs of other species, etc. The hsa-miR-4294 gene can be obtained by the method described in GoffLA et al., 2009, PLoS One, Vol. 4, e7192. In addition, as a precursor of "hsa-miR-4294", "hsa-mir-4294" (miRBase accession number MI0015827, sequence number 153) is known to form a hairpin-like structure.

[0102] The terms "hsa-miR-150-3p gene" or "hsa-miR-150-3p" as used in this specification include the hsa-miR-150-3p gene described in sequence number 6 (miRBase accession number MIMAT0004610), homologs or orthologs of other species, etc. The hsa-miR-150-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12, pp. 735-739. In addition, as a precursor of "hsa-miR-150-3p", "hsa-mir-150" (miRBase accession number MI0000479, sequence number 154) is known to form a hairpin-like structure.

[0103] The terms "hsa-miR-6729-5p gene" or "hsa-miR-6729-5p" as used in this specification include the hsa-miR-6729-5p gene described in sequence number 7 (miRBase accession number MIMAT0027359), homologs or orthologs of other species, etc. The hsa-miR-6729-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6729-5p", "hsa-mir-6729" (miRBase accession number MI0022574, sequence number 155) is known to form a hairpin-like structure.

[0104] The terms "hsa-miR-7641 gene" or "hsa-miR-7641" as used in this specification include the hsa-miR-7641 gene described in sequence number 8 (miRBase accession number MIMAT0029782), homologs or orthologs of other species, etc. The hsa-miR-7641 gene can be obtained by the method described in Yoo JK et al., 2013, Arch Pharm Res, Vol. 36, pp. 353-358. In addition, as precursors to "hsa-miR-7641", "hsa-mir-7641-1" and "hsa-mir-7641-2" (miRBase accession numbers MI0024975, MI0024976, sequence numbers 156, 157) are known to form hairpin-like structures.

[0105] The terms "hsa-miR-6765-3p gene" or "hsa-miR-6765-3p" as used in this specification include the hsa-miR-6765-3p gene described in sequence number 9 (miRBase accession number MIMAT0027431), homologs or orthologs of other species, etc. The hsa-miR-6765-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6765-3p", "hsa-mir-6765" (miRBase accession number MI0022610, sequence number 158) is known to form a hairpin-like structure.

[0106] The terms "hsa-miR-6820-5p gene" or "hsa-miR-6820-5p" as used in this specification include the hsa-miR-6820-5p gene described in sequence number 10 (miRBase accession number MIMAT0027540), homologs or orthologs of other species, etc. The hsa-miR-6820-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6820-5p", "hsa-mir-6820" (miRBase accession number MI0022665, sequence number 159) is known to form a hairpin-like structure.

[0107] The terms "hsa-miR-575 gene" or "hsa-miR-575" as used in this specification include the hsa-miR-575 gene described in sequence number 11 (miRBase accession number MIMAT0003240), homologs or orthologs of other species, etc. The hsa-miR-575 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-575", "hsa-mir-575" (miRBase accession number MI0003582, sequence number 160) is known to form a hairpin-like structure.

[0108] The terms "hsa-miR-6836-3p gene" or "hsa-miR-6836-3p" as used in this specification include the hsa-miR-6836-3p gene described in sequence number 12 (miRBase accession number MIMAT0027575), homologs or orthologs of other species, etc. The hsa-miR-6836-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6836-3p", "hsa-mir-6836" (miRBase accession number MI0022682, sequence number 161) is known to form a hairpin-like structure.

[0109] The terms "hsa-miR-1469 gene" or "hsa-miR-1469" as used in this specification include the hsa-miR-1469 gene described in sequence number 13 (miRBase accession number MIMAT0007347), homologs or orthologs of other species, etc. The hsa-miR-1469 gene can be obtained by the method described in Kawaji H et al., 2008, BMC Genomics, Vol. 9, p. 157. In addition, as a precursor of "hsa-miR-1469", "hsa-mir-1469" (miRBase accession number MI0007074, sequence number 162) that forms a hairpin-like structure is known.

[0110] The terms "hsa-miR-663a gene" or "hsa-miR-663a" as used in this specification include the hsa-miR-663a gene described in sequence number 14 (miRBase accession number MIMAT0003326), homologs or orthologs of other species, etc. The hsa-miR-663a gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci U SA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-663a", "hsa-mir-663a" (miRBase accession number MI0003672, sequence number 163) is known to form a hairpin-like structure.

[0111] The terms "hsa-miR-6075 gene" or "hsa-miR-6075" as used in this specification include the hsa-miR-6075 gene described in sequence number 15 (miRBase accession number MIMAT0023700), homologs or orthologs of other species, etc. The hsa-miR-6075 gene can be obtained using the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor to "hsa-miR-6075," "hsa-mir-6075" (miRBase accession number MI0020352, sequence number 164), which forms a hairpin-like structure, is known.

[0112] The terms "hsa-miR-4634 gene" or "hsa-miR-4634" as used in this specification include the hsa-miR-4634 gene described in sequence number 16 (miRBase accession number MIMAT0019691), homologs or orthologs of other species, etc. The hsa-miR-4634 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4634", "hsa-mir-4634" (miRBase accession number MI0017261, sequence number 165) is known to form a hairpin-like structure.

[0113] The terms "hsa-miR-423-5p gene" or "hsa-miR-423-5p" as used in this specification include the hsa-miR-423-5p gene described in sequence number 17 (miRBase accession number MIMAT0004748), homologs or orthologs of other species, etc. The hsa-miR-423-5p gene can be obtained by the method described in Kasashima K et al., 2004, Biochem Biophys Res Commun., Vol. 322, pp. 403-410. In addition, as a precursor of "hsa-miR-423-5p", "hsa-mir-423" (miRBase accession number MI0001445, sequence number 166) is known to form a hairpin-like structure.

[0114] The terms "hsa-miR-4454 gene" or "hsa-miR-4454" as used in this specification include the hsa-miR-4454 gene described in sequence number 18 (miRBase accession number MIMAT0018976), homologs or orthologs of other species, etc. The hsa-miR-4454 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4454", "hsa-mir-4454" (miRBase accession number MI0016800, sequence number 167) is known to form a hairpin-like structure.

[0115] The terms "hsa-miR-7109-5p gene" or "hsa-miR-7109-5p" as used in this specification include the hsa-miR-7109-5p gene described in sequence number 19 (miRBase accession number MIMAT0028115), homologs or orthologs of other species, etc. The hsa-miR-7109-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-7109-5p", "hsa-mir-7109" (miRBase accession number MI0022960, sequence number 168) is known to form a hairpin-like structure.

[0116] The terms "hsa-miR-6789-5p gene" or "hsa-miR-6789-5p" as used in this specification include the hsa-miR-6789-5p gene described in sequence number 20 (miRBase accession number MIMAT0027478), homologs or orthologs of other species, etc. The hsa-miR-6789-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6789-5p", "hsa-mir-6789" (miRBase accession number MI0022634, sequence number 169) is known to form a hairpin-like structure.

[0117] The terms "hsa-miR-6877-5p gene" or "hsa-miR-6877-5p" as used in this specification include the hsa-miR-6877-5p gene described in sequence number 21 (miRBase accession number MIMAT0027654), homologs or orthologs of other species, etc. The hsa-miR-6877-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6877-5p", "hsa-mir-6877" (miRBase accession number MI0022724, sequence number 170) is known to form a hairpin-like structure.

[0118] The terms "hsa-miR-4792 gene" or "hsa-miR-4792" as used in this specification include the hsa-miR-4792 gene described in sequence number 22 (miRBase accession number MIMAT0019964), homologs or orthologs of other species, etc. The hsa-miR-4792 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4792", "hsa-mir-4792" (miRBase accession number MI0017439, sequence number 171) is known to form a hairpin-like structure.

[0119] The terms "hsa-miR-4530 gene" or "hsa-miR-4530" as used in this specification include the hsa-miR-4530 gene described in sequence number 23 (miRBase accession number MIMAT0019069), homologs or orthologs of other species, etc. The hsa-miR-4530 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4530", "hsa-mir-4530" (miRBase accession number MI0016897, sequence number 172) is known to form a hairpin-like structure.

[0120] The terms "hsa-miR-7975 gene" or "hsa-miR-7975" as used in this specification include the hsa-miR-7975 gene described in sequence number 24 (miRBase accession number MIMAT0031178), homologs or orthologs of other species, etc. The hsa-miR-7975 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online edition. In addition, as a precursor of "hsa-miR-7975", "hsa-mir-7975" (miRBase accession number MI0025751, sequence number 173) forming a hairpin-like structure is known.

[0121] The terms "hsa-miR-6724-5p gene" or "hsa-miR-6724-5p" as used in this specification include the hsa-miR-6724-5p gene described in sequence number 25 (miRBase accession number MIMAT0025856), homologs or orthologs of other species, etc. The hsa-miR-6724-5p gene can be obtained by the method described in Li Y et al., 2012, Gene, Vol. 497, pp. 330-335. Furthermore, as a precursor to "hsa-miR-6724-5p", "hsa-mir-6724" (miRBase accession number MI0022559, sequence number 174), which forms a hairpin-like structure, is known.

[0122] The terms "hsa-miR-8073 gene" or "hsa-miR-8073" as used in this specification include the hsa-miR-8073 gene described in sequence number 26 (miRBase accession number MIMAT0031000), homologs or orthologs of other species, etc. The hsa-miR-8073 gene can be obtained using the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. Furthermore, as a precursor to "hsa-miR-8073," "hsa-mir-8073" (miRBase accession number MI0025909, sequence number 175) is known to form a hairpin-like structure.

[0123] The terms "hsa-miR-7977 gene" or "hsa-miR-7977" as used in this specification include the hsa-miR-7977 gene described in sequence number 27 (miRBase accession number MIMAT0031180), homologs or orthologs of other species, etc. The hsa-miR-7977 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online edition. In addition, as a precursor of "hsa-miR-7977", "hsa-mir-7977" (miRBase accession number MI0025753, sequence number 176) is known to form a hairpin-like structure.

[0124] The terms "hsa-miR-1231 gene" or "hsa-miR-1231" as used in this specification include the hsa-miR-1231 gene described in sequence number 28 (miRBase accession number MIMAT0005586), homologs or orthologs of other species, etc. The hsa-miR-1231 gene can be obtained using the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor to "hsa-miR-1231," the hairpin-like structure "hsa-mir-1231" (miRBase accession number MI0006321, sequence number 177) is known.

[0125] The terms "hsa-miR-6799-5p gene" or "hsa-miR-6799-5p" as used in this specification include the hsa-miR-6799-5p gene described in sequence number 29 (miRBase accession number MIMAT0027498), homologs or orthologs of other species, etc. The hsa-miR-6799-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6799-5p", "hsa-mir-6799" (miRBase accession number MI0022644, sequence number 178) is known to form a hairpin-like structure.

[0126] The terms "hsa-miR-615-5p gene" or "hsa-miR-615-5p" as used in this specification include the hsa-miR-615-5p gene described in sequence number 30 (miRBase accession number MIMAT0004804), homologs or orthologs of other species, etc. The hsa-miR-615-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl AcadSci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-615-5p", "hsa-mir-615" (miRBase accession number MI0003628, sequence number 179), which forms a hairpin-like structure, is known.

[0127] The terms "hsa-miR-4450 gene" or "hsa-miR-4450" as used in this specification include the hsa-miR-4450 gene described in sequence number 31 (miRBase accession number MIMAT0018971), homologs or orthologs of other species, etc. The hsa-miR-4450 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4450", "hsa-mir-4450" (miRBase accession number MI0016795, sequence number 180) is known to form a hairpin-like structure.

[0128] The terms "hsa-miR-6726-5p gene" or "hsa-miR-6726-5p" as used in this specification include the hsa-miR-6726-5p gene described in sequence number 32 (miRBase accession number MIMAT0027353), homologs or orthologs of other species, etc. The hsa-miR-6726-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6726-5p", "hsa-mir-6726" (miRBase accession number MI0022571, sequence number 181) is known to form a hairpin-like structure.

[0129] The terms "hsa-miR-6875-5p gene" or "hsa-miR-6875-5p" as used in this specification include the hsa-miR-6875-5p gene described in sequence number 33 (miRBase accession number MIMAT0027650), homologs or orthologs of other species, etc. The hsa-miR-6875-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6875-5p", "hsa-mir-6875" (miRBase accession number MI0022722, sequence number 182) is known to form a hairpin-like structure.

[0130] The terms "hsa-miR-4734 gene" or "hsa-miR-4734" as used in this specification include the hsa-miR-4734 gene described in sequence number 34 (miRBase accession number MIMAT0019859), homologs or orthologs of other species, etc. The hsa-miR-4734 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4734", "hsa-mir-4734" (miRBase accession number MI0017371, sequence number 183) is known to form a hairpin-like structure.

[0131] The terms "hsa-miR-16-5p gene" or "hsa-miR-16-5p" as used in this specification include the hsa-miR-16-5p gene described in sequence number 35 (miRBase accession number MIMAT0000069), homologs or orthologs of other species, etc. The hsa-miR-16-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2001, Science, Vol. 294, pp. 853-858. In addition, as precursors to "hsa-miR-16-5p", "hsa-mir-16-1" and "hsa-mir-16-2" (miRBase accession numbers MI0000070, MI0000115, sequence numbers 184, 185) are known to form hairpin-like structures.

[0132] The terms "hsa-miR-602 gene" or "hsa-miR-602" as used in this specification include the hsa-miR-602 gene described in sequence number 36 (miRBase accession number MIMAT0003270), homologs or orthologs of other species, etc. The hsa-miR-602 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-602", "hsa-mir-602" (miRBase accession number MI0003615, sequence number 186) is known to form a hairpin-like structure.

[0133] The terms "hsa-miR-4651 gene" or "hsa-miR-4651" as used in this specification include the hsa-miR-4651 gene described in sequence number 37 (miRBase accession number MIMAT0019715), homologs or orthologs of other species, etc. The hsa-miR-4651 gene can be obtained using the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4651", "hsa-mir-4651" (miRBase accession number MI0017279, sequence number 187) is known to form a hairpin-like structure.

[0134] The terms "hsa-miR-8069 gene" or "hsa-miR-8069" as used in this specification include the hsa-miR-8069 gene described in sequence number 38 (miRBase accession number MIMAT0030996), homologs or orthologs of other species, etc. The hsa-miR-8069 gene can be obtained using the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. Furthermore, as a precursor to "hsa-miR-8069", "hsa-mir-8069" (miRBase accession number MI0025905, sequence number 188) is known to form a hairpin-like structure.

[0135] The terms "hsa-miR-1238-5p gene" or "hsa-miR-1238-5p" as used in this specification include the hsa-miR-1238-5p gene described in sequence number 39 (miRBase accession number MIMAT0022947), homologs or orthologs of other species, etc. The hsa-miR-1238-5p gene can be obtained using the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor to "hsa-miR-1238-5p", "hsa-mir-1238" (miRBase accession number MI0006328, sequence number 189), which forms a hairpin-like structure, is known.

[0136] The terms "hsa-miR-6880-5p gene" or "hsa-miR-6880-5p" as used in this specification include the hsa-miR-6880-5p gene described in sequence number 40 (miRBase accession number MIMAT0027660), homologs or orthologs of other species, etc. The hsa-miR-6880-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6880-5p", "hsa-mir-6880" (miRBase accession number MI0022727, sequence number 190) is known to form a hairpin-like structure.

[0137] The terms "hsa-miR-8072 gene" or "hsa-miR-8072" as used in this specification include the hsa-miR-8072 gene described in sequence number 41 (miRBase accession number MIMAT0030999), homologs or orthologs of other species, etc. The hsa-miR-8072 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. Furthermore, as a precursor to "hsa-miR-8072", "hsa-mir-8072" (miRBase accession number MI0025908, sequence number 191) is known to form a hairpin-like structure.

[0138] The terms "hsa-miR-4723-5p gene" or "hsa-miR-4723-5p" as used in this specification include the hsa-miR-4723-5p gene described in sequence number 42 (miRBase accession number MIMAT0019838), homologs or orthologs of other species, etc. The hsa-miR-4723-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4723-5p", "hsa-mir-4723" (miRBase accession number MI0017359, sequence number 192) is known to form a hairpin-like structure.

[0139] The terms "hsa-miR-4732-5p gene" or "hsa-miR-4732-5p" as used in this specification include the hsa-miR-4732-5p gene described in sequence number 43 (miRBase accession number MIMAT0019855), homologs or orthologs of other species, etc. The hsa-miR-4732-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4732-5p", "hsa-mir-4732" (miRBase accession number MI0017369, sequence number 193) is known to form a hairpin-like structure.

[0140] The terms "hsa-miR-6125 gene" or "hsa-miR-6125" as used in this specification include the hsa-miR-6125 gene described in sequence number 44 (miRBase accession number MIMAT0024598), homologs or orthologs of other species, etc. The hsa-miR-6125 gene can be obtained by the method described in Smith JL et al., 2012, J Virol, Vol. 86, pp. 5278-5287. Furthermore, as a precursor to "hsa-miR-6125", "hsa-mir-6125" (miRBase accession number MI0021259, sequence number 194) is known to form a hairpin-like structure.

[0141] The terms "hsa-miR-6090 gene" or "hsa-miR-6090" as used in this specification include the hsa-miR-6090 gene described in sequence number 45 (miRBase accession number MIMAT0023715), homologs or orthologs of other species, etc. The hsa-miR-6090 gene can be obtained using the method described in Yoo JK et al., 2012, Stem Cells Dev, Vol. 21, pp. 2049-2057. Furthermore, as a precursor to "hsa-miR-6090," "hsa-mir-6090" (miRBase accession number MI0020367, sequence number 195), which forms a hairpin-like structure, is known.

[0142] The terms “hsa-miR-7114-5p gene” or “hsa-miR-7114-5p” as used in this specification include the hsa-miR-7114-5p gene described in sequence number 46 (miRBase accession number MIMAT0028125), homologs or orthologs of other species, etc. The hsa-miR-7114-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of “hsa-miR-7114-5p”, “hsa-mir-7114” (miRBase accession number MI0022965, sequence number 196) which forms a hairpin-like structure is known.

[0143] The terms "hsa-miR-564 gene" or "hsa-miR-564" as used in this specification include the hsa-miR-564 gene described in sequence number 47 (miRBase accession number MIMAT0003228), homologs or orthologs of other species, etc. The hsa-miR-564 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-564," "hsa-mir-564" (miRBase accession number MI0003570, sequence number 197) is known to form a hairpin-like structure.

[0144] The terms "hsa-miR-451a gene" or "hsa-miR-451a" as used in this specification include the hsa-miR-451a gene described in sequence number 48 (miRBase accession number MIMAT0001631), homologs or orthologs of other species, etc. The hsa-miR-451a gene can be obtained by the method described in Altuvia Y et al., 2005, Nucleic Acids Res, Vol. 33, pp. 2697-2706. Furthermore, as a precursor to "hsa-miR-451a", "hsa-mir-451a" (miRBase accession number MI0001729, sequence number 198) is known to form a hairpin-like structure.

[0145] The terms "hsa-miR-3135b gene" or "hsa-miR-3135b" as used in this specification include the hsa-miR-3135b gene described in sequence number 49 (miRBase accession number MIMAT0018985), homologs or orthologs of other species, etc. The hsa-miR-3135b gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-3135b", "hsa-mir-3135b" (miRBase accession number MI0016809, sequence number 199) is known to form a hairpin-like structure.

[0146] The terms "hsa-miR-4497 gene" or "hsa-miR-4497" as used in this specification include the hsa-miR-4497 gene described in sequence number 50 (miRBase accession number MIMAT0019032), homologs or orthologs of other species, etc. The hsa-miR-4497 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4497", "hsa-mir-4497" (miRBase accession number MI0016859, sequence number 200) is known to form a hairpin-like structure.

[0147] The terms "hsa-miR-4665-5p gene" or "hsa-miR-4665-5p" as used in this specification include the hsa-miR-4665-5p gene described in sequence number 51 (miRBase accession number MIMAT0019739), homologs or orthologs of other species, etc. The hsa-miR-4665-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4665-5p", "hsa-mir-4665" (miRBase accession number MI0017295, sequence number 201) is known to form a hairpin-like structure.

[0148] The terms "hsa-miR-3622a-5p gene" or "hsa-miR-3622a-5p" as used in this specification include the hsa-miR-3622a-5p gene described in sequence number 52 (miRBase accession number MIMAT0018003), homologs or orthologs of other species, etc. The hsa-miR-3622a-5p gene can be obtained by the method described in Witten D et al., 2010, BMCBiol, Vol. 8, p. 58. In addition, as a precursor of "hsa-miR-3622a-5p", "hsa-mir-3622a" (miRBase accession number MI0016013, sequence number 202) is known to form a hairpin-like structure.

[0149] The terms "hsa-miR-6850-5p gene" or "hsa-miR-6850-5p" as used in this specification include the hsa-miR-6850-5p gene described in sequence number 53 (miRBase accession number MIMAT0027600), homologs or orthologs of other species, etc. The hsa-miR-6850-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6850-5p", "hsa-mir-6850" (miRBase accession number MI0022696, sequence number 203) is known to form a hairpin-like structure.

[0150] The terms "hsa-miR-6821-5p gene" or "hsa-miR-6821-5p" as used in this specification include the hsa-miR-6821-5p gene described in sequence number 54 (miRBase accession number MIMAT0027542), homologs or orthologs of other species, etc. The hsa-miR-6821-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6821-5p", "hsa-mir-6821" (miRBase accession number MI0022666, sequence number 204) is known to form a hairpin-like structure.

[0151] The terms "hsa-miR-5100 gene" or "hsa-miR-5100" as used in this specification include the hsa-miR-5100 gene described in sequence number 55 (miRBase accession number MIMAT0022259), homologs or orthologs of other species, etc. The hsa-miR-5100 gene can be obtained by the method described in Tandon M et al., 2012, Oral Dis, Vol. 18, pp. 127-131. In addition, as a precursor of "hsa-miR-5100", "hsa-mir-5100" (miRBase accession number MI0019116, sequence number 205) is known to form a hairpin-like structure.

[0152] The terms "hsa-miR-6872-3p gene" or "hsa-miR-6872-3p" as used in this specification include the hsa-miR-6872-3p gene described in sequence number 56 (miRBase accession number MIMAT0027645), homologs or orthologs of other species, etc. The hsa-miR-6872-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6872-3p", "hsa-mir-6872" (miRBase accession number MI0022719, sequence number 206) is known to form a hairpin-like structure.

[0153] The terms "hsa-miR-4433-3p gene" or "hsa-miR-4433-3p" as used in this specification include the hsa-miR-4433-3p gene described in sequence number 57 (miRBase accession number MIMAT0018949), homologs or orthologs of other species, etc. The hsa-miR-4433-3p gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4433-3p", "hsa-mir-4433" (miRBase accession number MI0016773, sequence number 207) is known to form a hairpin-like structure.

[0154] The terms "hsa-miR-1227-5p gene" or "hsa-miR-1227-5p" as used in this specification include the hsa-miR-1227-5p gene described in sequence number 58 (miRBase accession number MIMAT0022941), homologs or orthologs of other species, etc. The hsa-miR-1227-5p gene can be obtained using the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor to "hsa-miR-1227-5p", "hsa-mir-1227" (miRBase accession number MI0006316, sequence number 208), which forms a hairpin-like structure, is known.

[0155] The terms "hsa-miR-3188 gene" or "hsa-miR-3188" as used in this specification include the hsa-miR-3188 gene described in sequence number 59 (miRBase accession number MIMAT0015070), homologs or orthologs of other species, etc. The hsa-miR-3188 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3188", "hsa-mir-3188" (miRBase accession number MI0014232, sequence number 209) is known to form a hairpin-like structure.

[0156] The terms "hsa-miR-7704 gene" or "hsa-miR-7704" as used in this specification include the hsa-miR-7704 gene described in sequence number 60 (miRBase accession number MIMAT0030019), homologs or orthologs of other species, etc. The hsa-miR-7704 gene can be obtained by the method described in Swaminathan S et al., 2013, Biochem Biophys ResCommun, Vol. 434, pp. 228-234. Furthermore, as a precursor to "hsa-miR-7704", "hsa-mir-7704" (miRBase accession number MI0025240, sequence number 210) is known to form a hairpin-like structure.

[0157] The terms "hsa-miR-3185 gene" or "hsa-miR-3185" as used in this specification include the hsa-miR-3185 gene described in sequence number 61 (miRBase accession number MIMAT0015065), homologs or orthologs of other species, etc. The hsa-miR-3185 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3185", "hsa-mir-3185" (miRBase accession number MI0014227, sequence number 211) is known to form a hairpin-like structure.

[0158] The terms "hsa-miR-1908-3p gene" or "hsa-miR-1908-3p" as used in this specification include the hsa-miR-1908-3p gene described in sequence number 62 (miRBase accession number MIMAT0026916), homologs or orthologs of other species, etc. The hsa-miR-1908-3p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, Vol. 26, pp. 2496-2505. Furthermore, as a precursor to "hsa-miR-1908-3p", "hsa-mir-1908" (miRBase accession number MI0008329, sequence number 212), which forms a hairpin-like structure, is known.

[0159] The terms "hsa-miR-6781-5p gene" or "hsa-miR-6781-5p" as used in this specification include the hsa-miR-6781-5p gene described in sequence number 63 (miRBase accession number MIMAT0027462), homologs or orthologs of other species, etc. The hsa-miR-6781-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6781-5p", "hsa-mir-6781" (miRBase accession number MI0022626, sequence number 213) is known to form a hairpin-like structure.

[0160] The terms "hsa-miR-6805-5p gene" or "hsa-miR-6805-5p" as used in this specification include the hsa-miR-6805-5p gene described in sequence number 64 (miRBase accession number MIMAT0027510), homologs or orthologs of other species, etc. The hsa-miR-6805-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6805-5p", "hsa-mir-6805" (miRBase accession number MI0022650, sequence number 214) is known to form a hairpin-like structure.

[0161] The terms "hsa-miR-8089 gene" or "hsa-miR-8089" as used in this specification include the hsa-miR-8089 gene described in sequence number 65 (miRBase accession number MIMAT0031016), homologs or orthologs of other species, etc. The hsa-miR-8089 gene can be obtained using the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. Furthermore, as a precursor to "hsa-miR-8089", "hsa-mir-8089" (miRBase accession number MI0025925, sequence number 215) is known to form a hairpin-like structure.

[0162] The terms "hsa-miR-665 gene" or "hsa-miR-665" as used in this specification include the hsa-miR-665 gene described in sequence number 66 (miRBase accession number MIMAT0004952), homologs or orthologs of other species, etc. The hsa-miR-665 gene can be obtained by the method described in Berezikov E et al., 2006, Genome Res, Vol. 16, pp. 1289-1298. Furthermore, as a precursor to "hsa-miR-665", "hsa-mir-665" (miRBase accession number MI0005563, sequence number 216) is known to form a hairpin-like structure.

[0163] The terms "hsa-miR-4486 gene" or "hsa-miR-4486" as used in this specification include the hsa-miR-4486 gene described in sequence number 67 (miRBase accession number MIMAT0019020), homologs or orthologs of other species, etc. The hsa-miR-4486 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4486", "hsa-mir-4486" (miRBase accession number MI0016847, sequence number 217) is known to form a hairpin-like structure.

[0164] The terms "hsa-miR-6722-3p gene" or "hsa-miR-6722-3p" as used in this specification include the hsa-miR-6722-3p gene described in sequence number 68 (miRBase accession number MIMAT0025854), homologs or orthologs of other species, etc. The hsa-miR-6722-3p gene can be obtained by the method described in Li Y et al., 2012, Gene, Vol. 497, pp. 330-335. Furthermore, as a precursor to "hsa-miR-6722-3p", "hsa-mir-6722" (miRBase accession number MI0022557, sequence number 218), which forms a hairpin-like structure, is known.

[0165] The terms "hsa-miR-1260a gene" or "hsa-miR-1260a" as used in this specification include the hsa-miR-1260a gene described in sequence number 69 (miRBase accession number MIMAT0005911), homologs or orthologs of other species, etc. The hsa-miR-1260a gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, Vol. 18, pp. 610-621. In addition, as a precursor of "hsa-miR-1260a", "hsa-mir-1260a" (miRBase accession number MI0006394, sequence number 219) is known to form a hairpin-like structure.

[0166] The terms "hsa-miR-4707-5p gene" or "hsa-miR-4707-5p" as used in this specification include the hsa-miR-4707-5p gene described in sequence number 70 (miRBase accession number MIMAT0019807), homologs or orthologs of other species, etc. The hsa-miR-4707-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4707-5p", "hsa-mir-4707" (miRBase accession number MI0017340, sequence number 220) is known to form a hairpin-like structure.

[0167] The terms "hsa-miR-6741-5p gene" or "hsa-miR-6741-5p" as used in this specification include the hsa-miR-6741-5p gene described in sequence number 71 (miRBase accession number MIMAT0027383), homologs or orthologs of other species, etc. The hsa-miR-6741-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6741-5p", "hsa-mir-6741" (miRBase accession number MI0022586, sequence number 221) is known to form a hairpin-like structure.

[0168] The terms "hsa-miR-1260b gene" or "hsa-miR-1260b" as used in this specification include the hsa-miR-1260b gene described in sequence number 72 (miRBase accession number MIMAT0015041), homologs or orthologs of other species, etc. The hsa-miR-1260b gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. In addition, as a precursor of "hsa-miR-1260b", "hsa-mir-1260b" (miRBase accession number MI0014197, sequence number 222) is known to form a hairpin-like structure.

[0169] The terms "hsa-miR-1246 gene" or "hsa-miR-1246" as used in this specification include the hsa-miR-1246 gene described in sequence number 73 (miRBase accession number MIMAT0005898), homologs or orthologs of other species, etc. The hsa-miR-1246 gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, Vol. 18, pp. 610-621. In addition, as a precursor of "hsa-miR-1246", "hsa-mir-1246" (miRBase accession number MI0006381, sequence number 223) is known to form a hairpin-like structure.

[0170] The terms "hsa-miR-6845-5p gene" or "hsa-miR-6845-5p" as used in this specification include the hsa-miR-6845-5p gene described in sequence number 74 (miRBase accession number MIMAT0027590), homologs or orthologs of other species, etc. The hsa-miR-6845-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6845-5p", "hsa-mir-6845" (miRBase accession number MI0022691, sequence number 224) is known to form a hairpin-like structure.

[0171] The terms "hsa-miR-4638-5p gene" or "hsa-miR-4638-5p" as used in this specification include the hsa-miR-4638-5p gene described in sequence number 75 (miRBase accession number MIMAT0019695), homologs or orthologs of other species, etc. The hsa-miR-4638-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4638-5p", "hsa-mir-4638" (miRBase accession number MI0017265, sequence number 225) is known to form a hairpin-like structure.

[0172] The terms "hsa-miR-6085 gene" or "hsa-miR-6085" as used in this specification include the hsa-miR-6085 gene described in sequence number 76 (miRBase accession number MIMAT0023710), homologs or orthologs of other organisms, etc. The hsa-miR-6085 gene can be obtained using the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor to "hsa-miR-6085," "hsa-mir-6085" (miRBase accession number MI0020362, sequence number 226), which forms a hairpin-like structure, is known.

[0173] The terms "hsa-miR-1228-3p gene" or "hsa-miR-1228-3p" as used in this specification include the hsa-miR-1228-3p gene described in sequence number 77 (miRBase accession number MIMAT0005583), homologs or orthologs of other species, etc. The hsa-miR-1228-3p gene can be obtained using the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor to "hsa-miR-1228-3p", "hsa-mir-1228" (miRBase accession number MI0006318, sequence number 227), which forms a hairpin-like structure, is known.

[0174] The terms "hsa-miR-4534 gene" or "hsa-miR-4534" as used in this specification include the hsa-miR-4534 gene described in sequence number 78 (miRBase accession number MIMAT0019073), homologs or orthologs of other species, etc. The hsa-miR-4534 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4534", "hsa-mir-4534" (miRBase accession number MI0016901, sequence number 228) is known to form a hairpin-like structure.

[0175] The terms "hsa-miR-5585-3p gene" or "hsa-miR-5585-3p" as used in this specification include the hsa-miR-5585-3p gene described in sequence number 79 (miRBase accession number MIMAT0022286), homologs or orthologs of other species, etc. The hsa-miR-5585-3p gene can be obtained by the method described in Friedlander MR et al., 2012, Nucleic Acids Res, Vol. 40, pp. 37-52. In addition, as a precursor of "hsa-miR-5585-3p", "hsa-mir-5585" (miRBase accession number MI0019142, sequence number 229) is known to form a hairpin-like structure.

[0176] The terms "hsa-miR-4741 gene" or "hsa-miR-4741" as used in this specification include the hsa-miR-4741 gene described in sequence number 80 (miRBase accession number MIMAT0019871), homologs or orthologs of other species, etc. The hsa-miR-4741 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4741", "hsa-mir-4741" (miRBase accession number MI0017379, sequence number 230) is known to form a hairpin-like structure.

[0177] The terms "hsa-miR-4433b-3p gene" or "hsa-miR-4433b-3p" as used in this specification include the hsa-miR-4433b-3p gene described in sequence number 81 (miRBase accession number MIMAT0030414), homologs or orthologs of other species, etc. The hsa-miR-4433b-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746. In addition, as a precursor of "hsa-miR-4433b-3p", "hsa-mir-4433b" (miRBase accession number MI0025511, sequence number 231) is known to form a hairpin-like structure.

[0178] The terms "hsa-miR-197-5p gene" or "hsa-miR-197-5p" as used in this specification include the hsa-miR-197-5p gene described in sequence number 82 (miRBase accession number MIMAT0022691), homologs or orthologs of other organisms, etc. The hsa-miR-197-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2003, RNA, Vol. 9, pp. 175-179. In addition, as a precursor of "hsa-miR-197-5p", "hsa-mir-197" (miRBase accession number MI0000239, sequence number 232) is known to form a hairpin-like structure.

[0179] The terms "hsa-miR-718 gene" or "hsa-miR-718" as used in this specification include the hsa-miR-718 gene described in sequence number 83 (miRBase accession number MIMAT0012735), homologs or orthologs of other species, etc. The hsa-miR-718 gene can be obtained by the method described in Artzi S et al., 2008, BMC Bioinformatics, Vol. 9, p39. In addition, as a precursor of "hsa-miR-718", "hsa-mir-718" (miRBase accession number MI0012489, sequence number 233) is known to form a hairpin-like structure.

[0180] The terms "hsa-miR-4513 gene" or "hsa-miR-4513" as used in this specification include the hsa-miR-4513 gene described in sequence number 84 (miRBase accession number MIMAT0019050), homologs or orthologs of other species, etc. The hsa-miR-4513 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4513", "hsa-mir-4513" (miRBase accession number MI0016879, sequence number 234) is known to form a hairpin-like structure.

[0181] The terms "hsa-miR-4446-3p gene" or "hsa-miR-4446-3p" as used in this specification include the hsa-miR-4446-3p gene described in sequence number 85 (miRBase accession number MIMAT0018965), homologs or orthologs of other species, etc. The hsa-miR-4446-3p gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4446-3p", "hsa-mir-4446" (miRBase accession number MI0016789, sequence number 235) is known to form a hairpin-like structure.

[0182] The terms "hsa-miR-619-5p gene" or "hsa-miR-619-5p" as used in this specification include the hsa-miR-619-5p gene described in sequence number 86 (miRBase accession number MIMAT0026622), homologs or orthologs of other species, etc. The hsa-miR-619-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl AcadSci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-619-5p", "hsa-mir-619" (miRBase accession number MI0003633, sequence number 236), which forms a hairpin-like structure, is known.

[0183] The terms "hsa-miR-6816-5p gene" or "hsa-miR-6816-5p" as used in this specification include the hsa-miR-6816-5p gene described in sequence number 87 (miRBase accession number MIMAT0027532), homologs or orthologs of other species, etc. The hsa-miR-6816-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6816-5p", "hsa-mir-6816" (miRBase accession number MI0022661, sequence number 237) is known to form a hairpin-like structure.

[0184] The terms "hsa-miR-6778-5p gene" or "hsa-miR-6778-5p" as used in this specification include the hsa-miR-6778-5p gene described in sequence number 88 (miRBase accession number MIMAT0027456), homologs or orthologs of other species, etc. The hsa-miR-6778-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6778-5p", "hsa-mir-6778" (miRBase accession number MI0022623, sequence number 238), which forms a hairpin-like structure, is known.

[0185] The terms "hsa-miR-24-3p gene" or "hsa-miR-24-3p" as used in this specification include the hsa-miR-24-3p gene described in sequence number 89 (miRBase accession number MIMAT0000080), homologs or orthologs of other species, etc. The hsa-miR-24-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2001, Science, Vol. 294, pp. 853-858. In addition, as precursors to "hsa-miR-24-3p", "hsa-mir-24-1" and "hsa-mir-24-2" (miRBase accession numbers MI0000080, MI0000081, sequence numbers 239, 240) are known to form hairpin-like structures.

[0186] The terms "hsa-miR-1915-3p gene" or "hsa-miR-1915-3p" as used in this specification include the hsa-miR-1915-3p gene described in sequence number 90 (miRBase accession number MIMAT0007892), homologs or orthologs of other species, etc. The hsa-miR-1915-3p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, Vol. 26, pp. 2496-2505. Furthermore, as a precursor to "hsa-miR-1915-3p", "hsa-mir-1915" (miRBase accession number MI0008336, sequence number 241), which forms a hairpin-like structure, is known.

[0187] The terms "hsa-miR-4665-3p gene" or "hsa-miR-4665-3p" as used in this specification include the hsa-miR-4665-3p gene described in sequence number 91 (miRBase accession number MIMAT0019740), homologs or orthologs of other species, etc. The hsa-miR-4665-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4665-3p", "hsa-mir-4665" (miRBase accession number MI0017295, sequence number 201) is known to form a hairpin-like structure.

[0188] The terms "hsa-miR-4449 gene" or "hsa-miR-4449" as used in this specification include the hsa-miR-4449 gene described in sequence number 92 (miRBase accession number MIMAT0018968), homologs or orthologs of other species, etc. The hsa-miR-4449 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4449", "hsa-mir-4449" (miRBase accession number MI0016792, sequence number 242) is known to form a hairpin-like structure.

[0189] The terms "hsa-miR-6889-5p gene" or "hsa-miR-6889-5p" as used in this specification include the hsa-miR-6889-5p gene described in sequence number 93 (miRBase accession number MIMAT0027678), homologs or orthologs of other species, etc. The hsa-miR-6889-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6889-5p", "hsa-mir-6889" (miRBase accession number MI0022736, sequence number 243), which forms a hairpin-like structure, is known.

[0190] The terms "hsa-miR-486-3p gene" or "hsa-miR-486-3p" as used in this specification include the hsa-miR-486-3p gene described in sequence number 94 (miRBase accession number MIMAT0004762), homologs or orthologs of other species, etc. The hsa-miR-486-3p gene can be obtained by the method described in Fu H et al., 2005, FEBS Lett, Vol. 579, pp. 3849-3854. In addition, as precursors to "hsa-miR-486-3p", "hsa-mir-486" and "hsa-mir-486-2" (miRBase accession numbers MI0002470 and MI0023622, sequence numbers 244 and 245) are known to form hairpin-like structures.

[0191] The terms "hsa-miR-7113-3p gene" or "hsa-miR-7113-3p" as used in this specification include the hsa-miR-7113-3p gene described in sequence number 95 (miRBase accession number MIMAT0028124), homologs or orthologs of other species, etc. The hsa-miR-7113-3p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-7113-3p", "hsa-mir-7113" (miRBase accession number MI0022964, sequence number 246) is known to form a hairpin-like structure.

[0192] The terms "hsa-miR-642a-3p gene" or "hsa-miR-642a-3p" as used in this specification include the hsa-miR-642a-3p gene described in sequence number 96 (miRBase accession number MIMAT0020924), homologs or orthologs of other species, etc. The hsa-miR-642a-3p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-642a-3p", "hsa-mir-642a" (miRBase accession number MI0003657, sequence number 247), which forms a hairpin-like structure, is known.

[0193] The terms "hsa-miR-7847-3p gene" or "hsa-miR-7847-3p" as used in this specification include the hsa-miR-7847-3p gene described in sequence number 97 (miRBase accession number MIMAT0030422), homologs or orthologs of other species, etc. The hsa-miR-7847-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746. In addition, as a precursor of "hsa-miR-7847-3p", "hsa-mir-7847" (miRBase accession number MI0025517, sequence number 248) is known to form a hairpin-like structure.

[0194] The terms "hsa-miR-6768-5p gene" or "hsa-miR-6768-5p" as used in this specification include the hsa-miR-6768-5p gene described in sequence number 98 (miRBase accession number MIMAT0027436), homologs or orthologs of other species, etc. The hsa-miR-6768-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-6768-5p", "hsa-mir-6768" (miRBase accession number MI0022613, sequence number 249) is known to form a hairpin-like structure.

[0195] The terms "hsa-miR-1290 gene" or "hsa-miR-1290" as used in this specification include the hsa-miR-1290 gene described in sequence number 99 (miRBase accession number MIMAT0005880), homologs or orthologs of other species, etc. The hsa-miR-1290 gene can be obtained by the method described in Morin RD et al., 2008, Genome Res, Vol. 18, pp. 610-621. In addition, as a precursor of "hsa-miR-1290", "hsa-mir-1290" (miRBase accession number MI0006352, sequence number 250) is known to form a hairpin-like structure.

[0196] The terms "hsa-miR-7108-5p gene" or "hsa-miR-7108-5p" as used in this specification include the hsa-miR-7108-5p gene described in sequence number 100 (miRBase accession number MIMAT0028113), homologs or orthologs of other species, etc. The hsa-miR-7108-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-7108-5p", "hsa-mir-7108" (miRBase accession number MI0022959, sequence number 251) is known to form a hairpin-like structure.

[0197] The terms "hsa-miR-92b-5p gene" or "hsa-miR-92b-5p" as used in this specification include the hsa-miR-92b-5p gene described in sequence number 101 (miRBase accession number MIMAT0004792), homologs or orthologs of other species, etc. The hsa-miR-92b-5p gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl AcadSci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-92b-5p", "hsa-mir-92b" (miRBase accession number MI0003560, sequence number 252), which forms a hairpin-like structure, is known.

[0198] The terms "hsa-miR-663b gene" or "hsa-miR-663b" as used in this specification include the hsa-miR-663b gene described in sequence number 102 (miRBase accession number MIMAT0005867), homologs or orthologs of other species, etc. The hsa-miR-663b gene can be obtained using the method described in Takada S et al., 2008, Leukemia, Vol. 22, pp. 1274-1278. Furthermore, as a precursor to "hsa-miR-663b," the hairpin-like structure "hsa-mir-663b" (miRBase accession number MI0006336, sequence number 253) is known.

[0199] The terms "hsa-miR-3940-5p gene" or "hsa-miR-3940-5p" as used in this specification include the hsa-miR-3940-5p gene described in sequence number 103 (miRBase accession number MIMAT0019229), homologs or orthologs of other species, etc. The hsa-miR-3940-5p gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, Vol. 5, e10563. In addition, as a precursor of "hsa-miR-3940-5p", "hsa-mir-3940" (miRBase accession number MI0016597, sequence number 254) is known to form a hairpin-like structure.

[0200] The terms "hsa-miR-4467 gene" or "hsa-miR-4467" as used in this specification include the hsa-miR-4467 gene described in sequence number 104 (miRBase accession number MIMAT0018994), homologs or orthologs of other species, etc. The hsa-miR-4467 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4467", "hsa-mir-4467" (miRBase accession number MI0016818, sequence number 255) is known to form a hairpin-like structure.

[0201] The terms "hsa-miR-6858-5p gene" or "hsa-miR-6858-5p" as used in this specification include the hsa-miR-6858-5p gene described in sequence number 105 (miRBase accession number MIMAT0027616), homologs or orthologs of other species, etc. The hsa-miR-6858-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6858-5p", "hsa-mir-6858" (miRBase accession number MI0022704, sequence number 256), which forms a hairpin-like structure, is known.

[0202] The terms "hsa-miR-4417 gene" or "hsa-miR-4417" as used in this specification include the hsa-miR-4417 gene described in sequence number 106 (miRBase accession number MIMAT0018929), homologs or orthologs of other species, etc. The hsa-miR-4417 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4417", "hsa-mir-4417" (miRBase accession number MI0016753, sequence number 257) is known to form a hairpin-like structure.

[0203] The terms "hsa-miR-3665 gene" or "hsa-miR-3665" as used in this specification include the hsa-miR-3665 gene described in sequence number 107 (miRBase accession number MIMAT0018087), homologs or orthologs of other species, etc. The hsa-miR-3665 gene can be obtained using the method described by Xie X et al., 2005, Nature, Vol. 434, pp. 338-345. Furthermore, as a precursor to "hsa-miR-3665," the hairpin-like structure "hsa-mir-3665" (miRBase accession number MI0016066, sequence number 258) is known.

[0204] The terms "hsa-miR-4736 gene" or "hsa-miR-4736" as used in this specification include the hsa-miR-4736 gene described in sequence number 108 (miRBase accession number MIMAT0019862), homologs or orthologs of other species, etc. The hsa-miR-4736 gene can be obtained using the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4736," "hsa-mir-4736" (miRBase accession number MI0017373, sequence number 259), which forms a hairpin-like structure, is known.

[0205] The terms "hsa-miR-4687-3p gene" or "hsa-miR-4687-3p" as used in this specification include the hsa-miR-4687-3p gene described in sequence number 109 (miRBase accession number MIMAT0019775), homologs or orthologs of other species, etc. The hsa-miR-4687-3p gene can be obtained using the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4687-3p", "hsa-mir-4687" (miRBase accession number MI0017319, sequence number 260), which forms a hairpin-like structure, is known.

[0206] The terms "hsa-miR-1908-5p gene" or "hsa-miR-1908-5p" as used in this specification include the hsa-miR-1908-5p gene described in sequence number 110 (miRBase accession number MIMAT0007881), homologs or orthologs of other species, etc. The hsa-miR-1908-5p gene can be obtained by the method described in Bar M et al., 2008, Stem Cells, Vol. 26, pp. 2496-2505. Furthermore, as a precursor to "hsa-miR-1908-5p", "hsa-mir-1908" (miRBase accession number MI0008329, sequence number 212), which forms a hairpin-like structure, is known.

[0207] The terms "hsa-miR-5195-3p gene" or "hsa-miR-5195-3p" as used in this specification include the hsa-miR-5195-3p gene described in sequence number 111 (miRBase accession number MIMAT0021127), homologs or orthologs of other species, etc. The hsa-miR-5195-3p gene can be obtained by the method described in Schotte D et al., 2011, Leukemia, Vol. 25, pp. 1389-1399. Furthermore, as a precursor to "hsa-miR-5195-3p", "hsa-mir-5195" (miRBase accession number MI0018174, sequence number 261), which forms a hairpin-like structure, is known.

[0208] The terms "hsa-miR-4286 gene" or "hsa-miR-4286" as used in this specification include the hsa-miR-4286 gene described in sequence number 112 (miRBase accession number MIMAT0016916), homologs or orthologs of other species, etc. The hsa-miR-4286 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One, Vol. 4, e7192. In addition, as a precursor of "hsa-miR-4286", "hsa-mir-4286" (miRBase accession number MI0015894, sequence number 262) is known to form a hairpin-like structure.

[0209] The terms "hsa-miR-3679-3p gene" or "hsa-miR-3679-3p" as used in this specification include the hsa-miR-3679-3p gene described in sequence number 113 (miRBase accession number MIMAT0018105), homologs or orthologs of other species, etc. The hsa-miR-3679-3p gene can be obtained by the method described in Creighton CJ et al., 2010, PLoSOne, Vol. 5, e9637. In addition, as a precursor of "hsa-miR-3679-3p", "hsa-mir-3679" (miRBase accession number MI0016080, sequence number 263) is known to form a hairpin-like structure.

[0210] The terms "hsa-miR-6791-5p gene" or "hsa-miR-6791-5p" as used in this specification include the hsa-miR-6791-5p gene described in sequence number 114 (miRBase accession number MIMAT0027482), homologs or orthologs of other species, etc. The hsa-miR-6791-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6791-5p", "hsa-mir-6791" (miRBase accession number MI0022636, sequence number 264), which forms a hairpin-like structure, is known.

[0211] The terms "hsa-miR-1202 gene" or "hsa-miR-1202" as used in this specification include the hsa-miR-1202 gene described in sequence number 115 (miRBase accession number MIMAT0005865), homologs or orthologs of other species, etc. The hsa-miR-1202 gene can be obtained by the method described in Marton S et al., 2008, Leukemia, Vol. 22, pp. 330-338. Furthermore, as a precursor to "hsa-miR-1202", "hsa-mir-1202" (miRBase accession number MI0006334, sequence number 265) is known to form a hairpin-like structure.

[0212] The terms "hsa-miR-3656 gene" or "hsa-miR-3656" as used in this specification include the hsa-miR-3656 gene described in sequence number 116 (miRBase accession number MIMAT0018076), homologs or orthologs of other species, etc. The hsa-miR-3656 gene can be obtained by the method described in Meiri E et al., 2010, Nucleic Acids Res, Vol. 38, pp. 6234-6246. Furthermore, as a precursor to "hsa-miR-3656," the hairpin-like structure "hsa-mir-3656" (miRBase accession number MI0016056, sequence number 266) is known.

[0213] The terms "hsa-miR-4746-3p gene" or "hsa-miR-4746-3p" as used in this specification include the hsa-miR-4746-3p gene described in sequence number 117 (miRBase accession number MIMAT0019881), homologs or orthologs of other species, etc. The hsa-miR-4746-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4746-3p", "hsa-mir-4746" (miRBase accession number MI0017385, sequence number 267) is known to form a hairpin-like structure.

[0214] The terms "hsa-miR-3184-5p gene" or "hsa-miR-3184-5p" as used in this specification include the hsa-miR-3184-5p gene described in sequence number 118 (miRBase accession number MIMAT0015064), homologs or orthologs of other species, etc. The hsa-miR-3184-5p gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3184-5p", "hsa-mir-3184" (miRBase accession number MI0014226, sequence number 268) is known to form a hairpin-like structure.

[0215] The terms "hsa-miR-3937 gene" or "hsa-miR-3937" as used in this specification include the hsa-miR-3937 gene described in sequence number 119 (miRBase accession number MIMAT0018352), homologs or orthologs of other species, etc. The hsa-miR-3937 gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, Vol. 5, e10563. In addition, as a precursor of "hsa-miR-3937", "hsa-mir-3937" (miRBase accession number MI0016593, sequence number 269) is known to form a hairpin-like structure.

[0216] The terms "hsa-miR-6515-3p gene" or "hsa-miR-6515-3p" as used in this specification include the hsa-miR-6515-3p gene described in sequence number 120 (miRBase accession number MIMAT0025487), homologs or orthologs of other species, etc. The hsa-miR-6515-3p gene can be obtained by the method described in Joyce CE et al., 2011, Hum MolGenet, Vol. 20, pp. 4025-4040. In addition, as a precursor of "hsa-miR-6515-3p", "hsa-mir-6515" (miRBase accession number MI0022227, sequence number 270) is known to form a hairpin-like structure.

[0217] The terms "hsa-miR-6132 gene" or "hsa-miR-6132" as used in this specification include the hsa-miR-6132 gene described in sequence number 121 (miRBase accession number MIMAT0024616), homologs or orthologs of other species, etc. The hsa-miR-6132 gene can be obtained by the method described in Dannemann M et al., 2012, Genome Biol Evol, Vol. 4, pp. 552-564. Furthermore, as a precursor to "hsa-miR-6132", "hsa-mir-6132" (miRBase accession number MI0021277, sequence number 271) is known to form a hairpin-like structure.

[0218] The terms "hsa-miR-187-5p gene" or "hsa-miR-187-5p" as used in this specification include the hsa-miR-187-5p gene described in sequence number 122 (miRBase accession number MIMAT0004561), homologs or orthologs of other species, etc. The hsa-miR-187-5p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. In addition, as a precursor of "hsa-miR-187-5p", "hsa-mir-187" (miRBase accession number MI0000274, sequence number 272), which forms a hairpin-like structure, is known.

[0219] The terms "hsa-miR-7111-5p gene" or "hsa-miR-7111-5p" as used in this specification include the hsa-miR-7111-5p gene described in sequence number 123 (miRBase accession number MIMAT0028119), homologs or orthologs of other species, etc. The hsa-miR-7111-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. In addition, as a precursor of "hsa-miR-7111-5p", "hsa-mir-7111" (miRBase accession number MI0022962, sequence number 273) is known to form a hairpin-like structure.

[0220] The terms "hsa-miR-5787 gene" or "hsa-miR-5787" as used in this specification include the hsa-miR-5787 gene described in sequence number 124 (miRBase accession number MIMAT0023252), homologs or orthologs of other species, etc. The hsa-miR-5787 gene can be obtained by the method described in Yoo H et al., 2011, Biochem Biophys Res Commun., Vol. 415, pp. 567-572. Furthermore, as a precursor to "hsa-miR-5787", "hsa-mir-5787" (miRBase accession number MI0019797, sequence number 274) is known to form a hairpin-like structure.

[0221] The terms "hsa-miR-6779-5p gene" or "hsa-miR-6779-5p" as used in this specification include the hsa-miR-6779-5p gene described in sequence number 125 (miRBase accession number MIMAT0027458), homologs or orthologs of other species, etc. The hsa-miR-6779-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6779-5p", "hsa-mir-6779" (miRBase accession number MI0022624, sequence number 275), which forms a hairpin-like structure, is known.

[0222] The terms "hsa-miR-6808-5p gene" or "hsa-miR-6808-5p" as used in this specification include the hsa-miR-6808-5p gene described in sequence number 126 (miRBase accession number MIMAT0027516), homologs or orthologs of other species, etc. The hsa-miR-6808-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6808-5p", "hsa-mir-6808" (miRBase accession number MI0022653, sequence number 276), which forms a hairpin-like structure, is known.

[0223] The terms "hsa-miR-6774-5p gene" or "hsa-miR-6774-5p" as used in this specification include the hsa-miR-6774-5p gene described in sequence number 127 (miRBase accession number MIMAT0027448), homologs or orthologs of other species, etc. The hsa-miR-6774-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6774-5p", "hsa-mir-6774" (miRBase accession number MI0022619, sequence number 277), which forms a hairpin-like structure, is known.

[0224] The terms "hsa-miR-4656 gene" or "hsa-miR-4656" as used in this specification include the hsa-miR-4656 gene described in sequence number 128 (miRBase accession number MIMAT0019723), homologs or orthologs of other species, etc. The hsa-miR-4656 gene can be obtained using the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4656," the hairpin-like structure "hsa-mir-4656" (miRBase accession number MI0017284, sequence number 278) is known.

[0225] The terms "hsa-miR-6806-5p gene" or "hsa-miR-6806-5p" as used in this specification include the hsa-miR-6806-5p gene described in sequence number 129 (miRBase accession number MIMAT0027512), homologs or orthologs of other species, etc. The hsa-miR-6806-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6806-5p", "hsa-mir-6806" (miRBase accession number MI0022651, sequence number 279), which forms a hairpin-like structure, is known.

[0226] The terms "hsa-miR-1233-5p gene" or "hsa-miR-1233-5p" as used in this specification include the hsa-miR-1233-5p gene described in sequence number 130 (miRBase accession number MIMAT0022943), homologs or orthologs of other species, etc. The hsa-miR-1233-5p gene can be obtained by the method described in Berezikov E et al., 2007, MolCell, Vol. 28, pp. 328-336. In addition, as precursors to "hsa-miR-1233-5p", "hsa-mir-1233-1" and "hsa-mir-1233-2" (miRBase accession numbers MI0006323, MI0015973, sequence numbers 280, 281) are known to form hairpin-like structures.

[0227] The terms "hsa-miR-328-5p gene" or "hsa-miR-328-5p" as used in this specification include the hsa-miR-328-5p gene described in sequence number 131 (miRBase accession number MIMAT0026486), homologs or orthologs of other species, etc. The hsa-miR-328-5p gene can be obtained by the method described in Kim J et al., 2004, Proc Natl Acad Sci U SA, Vol. 101, pp. 360-365. Furthermore, as a precursor to "hsa-miR-328-5p", "hsa-mir-328" (miRBase accession number MI0000804, sequence number 282), which forms a hairpin-like structure, is known.

[0228] The terms "hsa-miR-4674 gene" or "hsa-miR-4674" as used in this specification include the hsa-miR-4674 gene described in sequence number 132 (miRBase accession number MIMAT0019756), homologs or orthologs of other species, etc. The hsa-miR-4674 gene can be obtained using the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4674," "hsa-mir-4674" (miRBase accession number MI0017305, sequence number 283), which forms a hairpin-like structure, is known.

[0229] The terms "hsa-miR-2110 gene" or "hsa-miR-2110" as used in this specification include the hsa-miR-2110 gene described in sequence number 133 (miRBase accession number MIMAT0010133), homologs or orthologs of other species, etc. The hsa-miR-2110 gene can be obtained by the method described in Zhu JY et al., 2009, J Virol, Vol. 83, pp. 3333-3341. In addition, as a precursor of "hsa-miR-2110", "hsa-mir-2110" (miRBase accession number MI0010629, sequence number 284) is known to form a hairpin-like structure.

[0230] The terms "hsa-miR-6076 gene" or "hsa-miR-6076" as used in this specification include the hsa-miR-6076 gene described in sequence number 134 (miRBase accession number MIMAT0023701), homologs or orthologs of other organisms, etc. The hsa-miR-6076 gene can be obtained using the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor to "hsa-miR-6076," the hairpin-like structure "hsa-mir-6076" (miRBase accession number MI0020353, sequence number 285) is known.

[0231] The terms "hsa-miR-3619-3p gene" or "hsa-miR-3619-3p" as used in this specification include the hsa-miR-3619-3p gene described in sequence number 135 (miRBase accession number MIMAT0019219), homologs or orthologs of other species, etc. The hsa-miR-3619-3p gene can be obtained by the method described in Witten D et al., 2010, BMC Biol, Vol. 8, p58. In addition, as a precursor of "hsa-miR-3619-3p", "hsa-mir-3619" (miRBase accession number MI0016009, sequence number 286) is known to form a hairpin-like structure.

[0232] The terms "hsa-miR-92a-2-5p gene" or "hsa-miR-92a-2-5p" as used in this specification include the hsa-miR-92a-2-5p gene described in sequence number 136 (miRBase accession number MIMAT0004508), homologs or orthologs of other species, etc. The hsa-miR-92a-2-5p gene can be obtained by the method described in Mourelatos Z et al., 2002, Genes Dev, Vol. 16, pp. 720-728. Furthermore, as a precursor to "hsa-miR-92a-2-5p", "hsa-mir-92a-2" (miRBase accession number MI0000094, sequence number 287), which forms a hairpin-like structure, is known.

[0233] The terms "hsa-miR-128-1-5p gene" or "hsa-miR-128-1-5p" as used in this specification include the hsa-miR-128-1-5p gene described in sequence number 137 (miRBase accession number MIMAT0026477), homologs or orthologs of other species, etc. The hsa-miR-128-1-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12, pp. 735-739. In addition, as a precursor of "hsa-miR-128-1-5p", "hsa-mir-128-1" (miRBase accession number MI0000447, sequence number 288), which forms a hairpin-like structure, is known.

[0234] The terms "hsa-miR-638 gene" or "hsa-miR-638" as used in this specification include the hsa-miR-638 gene described in sequence number 138 (miRBase accession number MIMAT0003308), homologs or orthologs of other species, etc. The hsa-miR-638 gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, pp. 3687-3692. Furthermore, as a precursor to "hsa-miR-638," "hsa-mir-638" (miRBase accession number MI0003653, sequence number 289), which forms a hairpin-like structure, is known.

[0235] The terms "hsa-miR-2861 gene" or "hsa-miR-2861" as used in this specification include the hsa-miR-2861 gene described in sequence number 139 (miRBase accession number MIMAT0013802), homologs or orthologs of other species, etc. The hsa-miR-2861 gene can be obtained by the method described in Li H et al., 2009, J Clin Invest, Vol. 119, pp. 3666-3677. Furthermore, as a precursor to "hsa-miR-2861", "hsa-mir-2861" (miRBase accession number MI0013006, sequence number 290) is known to form a hairpin-like structure.

[0236] The terms "hsa-miR-371a-5p gene" or "hsa-miR-371a-5p" as used in this specification include the hsa-miR-371a-5p gene described in sequence number 140 (miRBase accession number MIMAT0004687), homologs or orthologs of other species, etc. The hsa-miR-371a-5p gene can be obtained by the method described in Suh MR et al., 2004, Dev Biol, Vol. 270, pp. 488-498. In addition, as a precursor of "hsa-miR-371a-5p", "hsa-mir-371a" (miRBase accession number MI0000779, sequence number 291), which forms a hairpin-like structure, is known.

[0237] The terms "hsa-miR-211-3p gene" or "hsa-miR-211-3p" as used in this specification include the hsa-miR-211-3p gene described in sequence number 141 (miRBase accession number MIMAT0022694), homologs or orthologs of other species, etc. The hsa-miR-211-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. In addition, as a precursor of "hsa-miR-211-3p", "hsa-mir-211" (miRBase accession number MI0000287, sequence number 292) is known to form a hairpin-like structure.

[0238] The terms "hsa-miR-1273g-3p gene" or "hsa-miR-1273g-3p" as used in this specification include the hsa-miR-1273g-3p gene described in sequence number 142 (miRBase accession number MIMAT0022742), homologs or orthologs of other species, etc. The hsa-miR-1273g-3p gene can be obtained by the method described in Reshmi G et al., 2011, Genomics, Vol. 97, pp. 333-340. Furthermore, as a precursor to "hsa-miR-1273g-3p", "hsa-mir-1273g" (miRBase accession number MI0018003, sequence number 293), which forms a hairpin-like structure, is known.

[0239] The terms "hsa-miR-1203 gene" or "hsa-miR-1203" as used in this specification include the hsa-miR-1203 gene described in sequence number 143 (miRBase accession number MIMAT0005866), homologs or orthologs of other species, etc. The hsa-miR-1203 gene can be obtained using the method described in Marton S et al., 2008, Leukemia, Vol. 22, pp. 330-338. Furthermore, as a precursor to "hsa-miR-1203", "hsa-mir-1203" (miRBase accession number MI0006335, sequence number 294) is known to form a hairpin-like structure.

[0240] The terms "hsa-miR-122-5p gene" or "hsa-miR-122-5p" as used in this specification include the hsa-miR-122-5p gene described in sequence number 144 (miRBase accession number MIMAT0000421), homologs or orthologs of other species, etc. The hsa-miR-122-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, CurrBiol, Vol. 12, pp. 735-739. In addition, as a precursor of "hsa-miR-122-5p", "hsa-mir-122" (miRBase accession number MI0000442, sequence number 295) is known to form a hairpin-like structure.

[0241] The terms "hsa-miR-4258 gene" or "hsa-miR-4258" as used in this specification include the hsa-miR-4258 gene described in sequence number 145 (miRBase accession number MIMAT0016879), homologs or orthologs of other species, etc. The hsa-miR-4258 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One, Vol. 4, e7192. In addition, as a precursor of "hsa-miR-4258", "hsa-mir-4258" (miRBase accession number MI0015857, sequence number 296) is known to form a hairpin-like structure.

[0242] The terms "hsa-miR-4484 gene" or "hsa-miR-4484" as used in this specification include the hsa-miR-4484 gene described in sequence number 146 (miRBase accession number MIMAT0019018), homologs or orthologs of other species, etc. The hsa-miR-4484 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. In addition, as a precursor of "hsa-miR-4484", "hsa-mir-4484" (miRBase accession number MI0016845, sequence number 297) is known to form a hairpin-like structure.

[0243] The terms "hsa-miR-4648 gene" or "hsa-miR-4648" as used in this specification include the hsa-miR-4648 gene described in sequence number 147 (miRBase accession number MIMAT0019710), homologs or orthologs of other species, etc. The hsa-miR-4648 gene can be obtained by the method described in Persson H et al., 2011, Cancer Res, Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-4648", "hsa-mir-4648" (miRBase accession number MI0017275, sequence number 298) is known to form a hairpin-like structure.

[0244] The terms "hsa-miR-6780b-5p gene" or "hsa-miR-6780b-5p" as used in this specification include the hsa-miR-6780b-5p gene described in sequence number 148 (miRBase accession number MIMAT0027572), homologs or orthologs of other species, etc. The hsa-miR-6780b-5p gene can be obtained by the method described in Ladewig E et al., 2012, GenomeRes, Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6780b-5p", "hsa-mir-6780b" (miRBase accession number MI0022681, sequence number 299), which forms a hairpin-like structure, is known.

[0245] The terms "hsa-miR-4516 gene" or "hsa-miR-4516" as used in this specification include the hsa-miR-4516 gene described in sequence number 466 (miRBase accession number MIMAT0019053), homologs or orthologs of other species, etc. The hsa-miR-4516 gene can be obtained by the method described in Jima DD et al., 2010, Blood. Vol. 116, e118-e127. Furthermore, as a precursor to "hsa-miR-4516", "hsa-mir-4516" (miRBase accession number MI0016882, sequence number 479) is known to form a hairpin-like structure.

[0246] The terms "hsa-miR-4649-5p gene" or "hsa-miR-4649-5p" as used in this specification include the hsa-miR-4649-5p gene described in sequence number 467 (miRBase accession number MIMAT0019711), homologs or orthologs of other species, etc. The hsa-miR-4649-5p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., Vol. 71, pp. 78-86. Furthermore, as a precursor to "hsa-miR-4649-5p", "hsa-mir-4649" (miRBase accession number MI0017276, sequence number 480), which forms a hairpin-like structure, is known.

[0247] The terms "hsa-miR-760 gene" or "hsa-miR-760" as used in this specification include the hsa-miR-760 gene described in sequence number 468 (miRBase accession number MIMAT0004957), homologs or orthologs of other species, etc. The hsa-miR-760 gene can be obtained by the method described in Berezikov E et al., 2006, Genome Res., Vol. 16, pp. 289-1298. Furthermore, as a precursor to "hsa-miR-760," "hsa-mir-760" (miRBase accession number MI0005567, sequence number 481), which forms a hairpin-like structure, is known.

[0248] The terms "hsa-miR-3162-5p gene" or "hsa-miR-3162-5p" as used in this specification include the hsa-miR-3162-5p gene described in sequence number 469 (miRBase accession number MIMAT0015036), homologs or orthologs of other species, etc. The hsa-miR-3162-5p gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3162-5p", "hsa-mir-3162" (miRBase accession number MI0014192, sequence number 482) is known to form a hairpin-like structure.

[0249] The terms "hsa-miR-3178 gene" or "hsa-miR-3178" as used in this specification include the hsa-miR-3178 gene described in sequence number 470 (miRBase accession number MIMAT0015055), homologs or orthologs of other species, etc. The hsa-miR-3178 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3178", "hsa-mir-3178" (miRBase accession number MI0014212, sequence number 483) that forms a hairpin-like structure is known.

[0250] The terms "hsa-miR-940 gene" or "hsa-miR-940" as used in this specification include the hsa-miR-940 gene described in sequence number 471 (miRBase accession number MIMAT0004983), homologs or orthologs of other species, etc. The hsa-miR-940 gene can be obtained using the method described in Lui WO et al., 2007, Cancer Res., Vol. 67, pp. 6031-6043. Furthermore, as a precursor to "hsa-miR-940," "hsa-mir-940" (miRBase accession number MI0005762, sequence number 484), which forms a hairpin-like structure, is known.

[0251] The terms "hsa-miR-4271 gene" or "hsa-miR-4271" as used in this specification include the hsa-miR-4271 gene described in sequence number 472 (miRBase accession number MIMAT0016901), homologs or orthologs of other species, etc. The hsa-miR-4271 gene can be obtained by the method described in Goff LA et al., 2009, PLoS One., Vol. 4, e7192. In addition, as a precursor of "hsa-miR-4271", "hsa-mir-4271" (miRBase accession number MI0015879, sequence number 485) is known to form a hairpin-like structure.

[0252] The terms "hsa-miR-6769b-5p gene" or "hsa-miR-6769b-5p" as used in this specification include the hsa-miR-6769b-5p gene described in sequence number 473 (miRBase accession number MIMAT0027620), homologs or orthologs of other species, etc. The hsa-miR-6769b-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res., Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6769b-5p", "hsa-mir-6769b" (miRBase accession number MI0022706, sequence number 486), which forms a hairpin-like structure, is known.

[0253] The terms "hsa-miR-4508 gene" or "hsa-miR-4508" as used in this specification include the hsa-miR-4508 gene described in sequence number 474 (miRBase accession number MIMAT0019045), homologs or orthologs of other species, etc. The hsa-miR-4508 gene can be obtained by the method described in Jima DD et al., 2010, Blood., Vol. 116, e118-e127. Furthermore, as a precursor to "hsa-miR-4508", "hsa-mir-4508" (miRBase accession number MI0016872, sequence number 487) is known to form a hairpin-like structure.

[0254] The terms "hsa-miR-6826-5p gene" or "hsa-miR-6826-5p" as used in this specification include the hsa-miR-6826-5p gene described in sequence number 475 (miRBase accession number MIMAT0027552), homologs or orthologs of other species, etc. The hsa-miR-6826-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res., Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6826-5p", "hsa-mir-6826" (miRBase accession number MI0022671, sequence number 488), which forms a hairpin-like structure, is known.

[0255] The terms "hsa-miR-6757-5p gene" or "hsa-miR-6757-5p" as used in this specification include the hsa-miR-6757-5p gene described in sequence number 476 (miRBase accession number MIMAT0027414), homologs or orthologs of other species, etc. The hsa-miR-6757-5p gene can be obtained by the method described in Ladewig E et al., 2012, Genome Res., Vol. 22, pp. 1634-1645. Furthermore, as a precursor to "hsa-miR-6757-5p", the hairpin-like structure "hsa-mir-6757" (miRBase accession number MI0022602, sequence number 489) is known.

[0256] The terms "hsa-miR-3131 gene" or "hsa-miR-3131" as used in this specification include the hsa-miR-3131 gene described in sequence number 477 (miRBase accession number MIMAT0014996), homologs or orthologs of other species, etc. The hsa-miR-3131 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. In addition, as a precursor of "hsa-miR-3131", "hsa-mir-3131" (miRBase accession number MI0014151, sequence number 490) is known to form a hairpin-like structure.

[0257] The terms "hsa-miR-1343-3p gene" or "hsa-miR-1343-3p" as used in this specification include the hsa-miR-1343-3p gene described in sequence number 478 (miRBase accession number MIMAT0019776), homologs or orthologs of other species, etc. The hsa-miR-1343-3p gene can be obtained by the method described in Persson H et al., 2011, Cancer Res., Vol. 71, pp. 78-86. In addition, as a precursor of "hsa-miR-1343-3p", "hsa-mir-1343" (miRBase accession number MI0017320, sequence number 491) is known to form a hairpin-like structure.

[0258] Furthermore, when mature miRNAs are excised from hairpin-like RNA precursors, sometimes one or more bases before and after the sequence are cut short or long, resulting in base substitutions and mutants called isomiRs (Morin RD. et al., 2008, Genome Research, Vol. 18, pp. 610-621). In miRBase version 20, in addition to the base sequences shown in any of the sequences 1-148 and 466-478, several mutants and fragments of isomiRs with base sequences shown in any of the sequences 300-465 and 492-509 are also shown. These mutants can also be obtained as miRNAs with base sequences shown in any of the sequences 1-148 and 466-478.

[0259] That is, the serial numbers 1, 3, 4, 6, 14, 16, 17, 18, 22, 23, 24, 25, 30, 31, 34, 35, 37, 42, 43, 44, 47, 48, 49, 50, 51, 52, 55, 57, 59, 61, 62, 66, 67, 69, 70, 72, 73, 75, 77, 79, 80, 82, 83, 84, 85, 86, 89, 90, 92, 94, 96, 99, 101, 102, 103, 104, 106, 107, 109, 1 of the present invention Among the polynucleotide mutants consisting of the base sequences shown in, e.g., 10, 111, 112, 113, 115, 116, 120, 121, 122, 124, 130, 131, 132, 133, 136, 137, 138, 139, 140, 141, 142, 144, 146, 147, 466, 467, 468, 469, 470, 471, 474, 477, and 478, or the polynucleotide mutants consisting of base sequences where u is t, the longest mutant is, for example, registered in miRBase version 20. The following serial numbers can be cited as examples: 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370, 372, 374, 376, 378, 380, 382, ​​384, 386, 388, 390. The polynucleotides are 392, 394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 492, 494, 496, 498, 500, 502, 504, 506, and 508.

[0260] Furthermore, the serial numbers 1, 3, 4, 6, 14, 16, 17, 18, 22, 23, 24, 25, 30, 31, 34, 35, 37, 42, 43, 44, 47, 48, 49, 50, 51, 52, 55, 57, 59, 61, 62, 66, 67, 69, 70, 72, 73, 75, 77, 79, 80, 82, 83, 84, 85, 86, 89, 90, 92, 94, 96, 99, 101, 102, 103, 104, 106, 107, 109, 1 of the present invention are... Among the polynucleotide mutants consisting of the base sequences shown in 10, 111, 112, 113, 115, 116, 120, 121, 122, 124, 130, 131, 132, 133, 136, 137, 138, 139, 140, 141, 142, 144, 146, 147, 466, 467, 468, 469, 470, 471, 474, 477, and 478, or the polynucleotide mutants consisting of base sequences where u is t, the shortest mutants registered, for example, in miRBase version 20, are classified as such. Other possible serial numbers include 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 3 The polynucleotide sequences shown are 93, 395, 397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463, 465, 493, 495, 497, 499, 501, 503, 505, 507, and 509.In addition to these mutants and fragments, other examples include those registered in miRBase as sequence numbers 1, 3, 4, 6, 14, 16, 17, 18, 22, 23, 24, 25, 30, 31, 34, 35, 37, 42, 43, 44, 47, 48, 49, 50, 51, 52, 55, 57, 59, 61, 62, 66, 67, 69, 70, 72, 73, 75, 77, 79, 80, 82, 83, 8... 4, 85, 86, 89, 90, 92, 94, 96, 99, 101, 102, 103, 104, 106, 107, 109, 110, 111, 112, 113, 115, 116, 120, 121, 122, 124, 130, 131, 132, 133, 136, 137, 138, 139, 140, 141, 142, 144, 146, and 147 are polynucleotides containing multiple isomiRs. Furthermore, as examples of polynucleotides containing the base sequences shown in any one of the sequence numbers 1-148 and 466-478, polynucleotides shown in any one of the sequence numbers 149-299 and 479-491 as precursors can be cited.

[0261] The names of the genes shown in sequence numbers 1 to 509 and their miRBase accession numbers are recorded in Table 1.

[0262] In this specification, "capable of specific binding" means that the nucleic acid probes or primers used in this invention bind to specific target nucleic acids, but cannot actually bind to other nucleic acids.

[0263] [Table 1]

[0264]

[0265]

[0266]

[0267]

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274]

[0275]

[0276]

[0277]

[0278]

[0279] This specification includes the contents described in the specifications and drawings of Japanese Patent Application Nos. 2014-120884 and 2014-185733, which form the basis of the priority of this application.

[0280] The effects of the invention

[0281] This invention enables the easy and highly accurate detection of biliary tract cancer. For example, by using the measurements of several miRNAs in the patient's blood, serum, and / or plasma, which can be collected in a minimally invasive manner, it is easy to determine whether the patient has biliary tract cancer. Attached Figure Description

[0282] Figure 1 The relationship between the base sequences of hsa-miR-4665-5p (as shown in sequence number 51) and hsa-miR-4665-3p (as shown in sequence number 91) generated from hsa-mir-4665 (as shown in sequence number 201) as a precursor is shown.

[0283] Figure 2 Left: A graph showing the hsa-miR-125a-3p (serial number 1) measurements in healthy individuals (100 people) and biliary tract cancer patients (67 people) selected as the learning specimen group, with the horizontal axis representing the values. The horizontal line in the graph represents the threshold (5.69) optimized by Fisher's discriminant analysis to distinguish between the two groups. Right: A graph showing the hsa-miR-125a-3p (serial number 1) measurements in healthy individuals (50 people) and biliary tract cancer patients (33 people) selected as the test specimen group, with the horizontal axis representing the threshold (5.69) set for distinguishing between the two groups in the learning specimen group.

[0284] Figure 3Left: A graph showing the hsa-miR-6893-5p (serial number 2) measurements on the x-axis and the hsa-miR-4476 (serial number 4) measurements on the y-axis for healthy individuals (100 people, circle) and biliary tract cancer patients (67 people, triangle) selected as the study specimen group. The lines in the graph represent the discriminant function (0 = 5.16x + y + 48.11) optimized by Fisher's discriminant analysis to distinguish between the two groups. Right: A graph showing the hsa-miR-6893-5p (serial number 2) measurements on the x-axis and the hsa-miR-4476 (serial number 4) measurements on the y-axis for healthy individuals (50 people, circle) and biliary tract cancer patients (33 people, triangle) selected as the test specimen group. The lines in the figure represent the thresholds set by the learning specimen group to distinguish between the two groups (0 = 5.16x + y + 48.11).

[0285] Figure 4 The image above shows the hsa-miR-6075 (serial number 15), hsa-miR-6836-3p (serial number 12), hsa-miR-6799-5p (serial number 29), and hsa-miR-1 samples from 67 patients with biliary tract cancer, 93 healthy individuals, 35 patients with colorectal cancer, 37 patients with gastric cancer, 32 patients with esophageal cancer, 38 patients with liver cancer, and 13 patients with benign pancreatic and biliary tract diseases selected as the study specimen group. The measured values ​​of 25a-3p (serial number 1) were analyzed using Fisher's discriminant analysis to generate a discriminant formula (-1.25×hsa-miR-6075-1.06×hsa-miR-6836-3p+0.53×hsa-miR-6799-5p+0.18×hsa-miR-125a-3p+15.41). A graph was plotted with the discriminant score obtained from this formula on the ordinate and the sample group on the x-axis. The dashed lines in the graph represent the discriminant boundary between the two groups where the discriminant score is 0. The following figure shows the hsa-miR-6075 (serial number 15), hsa-miR-6836-3p (serial number 12), hsa-miR-6799-5p (serial number 29), and hsa-miR-125a-3p (serial number 1) measurements for 33 patients with biliary tract cancer, 57 healthy individuals, 15 patients with colorectal cancer, 13 patients with gastric cancer, 18 patients with esophageal cancer, 12 patients with liver cancer, and 8 patients with benign pancreatic and biliary tract diseases, selected as the test specimen group. The graph is plotted with the discriminant score obtained from the discriminant expression based on the learning specimen group on the vertical axis and the specimen group on the horizontal axis. The dashed line in the figure represents the discriminant boundary between the two groups where the discriminant score is 0. Detailed Implementation

[0286] The present invention will be further described in detail below.

[0287] 1. Target nucleic acids for bile duct cancer

[0288] The primary target nucleic acids used for detecting the presence and / or absence of biliary tract cancer or biliary tract cancer cells using the nucleic acid probes or primers defined above in this invention, which serve as markers for biliary tract cancer, may be selected from hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR-150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, and hsa-miR-6836-3p. hsa-miR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-miR-423-5p, hsa-miR-4454, hsa-miR-7109-5p, hsa-miR-6789-5p, hsa-miR-6877 -5p, hsa-miR-4792, hsa-miR-4530, hsa-miR-7975, hsa-miR-6724-5p, hsa-miR-8073, hsa-miR-7977, hsa-miR-1231, hsa-miR-6799-5p, hsa-miR-615 -5p, hsa-miR-4450, hsa-miR-6726-5p, hsa-miR-6875-5p, hsa-miR-4734, hsa-miR-16-5p, hsa-miR-602, hsa-miR-4651, hsa-miR-8069, hsa-miR-12 38-5p, hsa-miR-6880-5p, hsa-miR-8072, hsa-miR-4723-5p, hsa-miR-4732-5p, hsa-miR-6125, hsa-miR-6090, hsa-miR-7114-5p, hsa-miR-564, hsa- miR-451a, hsa-miR-3135b, hsa-miR-4497, hsa-miR-4665-5p, hsa-miR-3622a-5p, hsa-miR-6850-5p, hsa-miR-6821-5p, hsa-miR-5100, hsa-miR-68 72-3p, hsa-miR-4433-3p, hsa-miR-1227-5p, hsa-miR-3188, hsa-miR-7704, hsa-miR-3185, hsa-miR-1908-3p, hsa-miR-6781-5p, hsa-miR-6805-5p,hsa-miR-8089, hsa-miR-665, hsa-miR-4486, hsa-miR-6722-3p, hsa-miR-1260a, hsa-miR-4707-5p, hsa-miR-6741-5p, hsa-miR-1260b, hsa-miR-12 46, hsa-miR-6845-5p, hsa-miR-4638-5p, hsa-miR-6085, hsa-miR-1228-3p, hsa-miR-4534, hsa-miR-5585-3p, hsa-miR-4741, hsa-miR-4433b-3p, h sa-miR-197-5p, hsa-miR-718, hsa-miR-4513, hsa-miR-4446-3p, hsa-miR-619-5p, hsa-miR-6816-5p, hsa-miR-6778-5p, hsa-miR-24-3p, hsa-miR- 1915-3p, hsa-miR-4665-3p, hsa-miR-4449, hsa-miR-6889-5p, hsa-miR-486-3p, hsa-miR-7113-3p, hsa-miR-642a-3p, hsa-miR-7847-3p, hsa-miR- 6768-5p, hsa-miR-1290, hsa-miR-7108-5p, hsa-miR-92b-5p, hsa-miR-663b, hsa-miR-3940-5p, hsa-miR-4467, hsa-miR-6858-5p, hsa-miR-4417, h sa-miR-3665, hsa-miR-4736, hsa-miR-4687-3p, hsa-miR-1908-5p, hsa-miR-5195-3p, hsa-miR-4286, hsa-miR-3679-3p, hsa-miR-6791-5p, hsa-miR-1202, hsa-miR-3656, hsa-miR-4746-3p, hsa-miR-3184-5p, hsa-miR-3937, hsa-miR-6515-3p, hsa-miR-6132, hsa-miR-187-5p, hsa-miR-7111-5p hsa-miR-5787 hsa-miR-6779-5p hsa-miR-4516 hsa-miR-4649-5p hsa-miR-760 hsa-miR-3162-5p hsa-miR-3178 hsa-miR-940 hsa-miR-4271At least one miRNA from hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-3131, and hsa-miR-1343-3p. Further, other bile duct cancer markers that can be combined with these miRNAs, namely, those selected from hsa-miR-6808-5p, hsa-miR-6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa-miR-1233-5p, hsa-miR-328-5p, hsa-miR-4674, hsa-miR-2110, hsa-miR-6076, hsa-miR-3619-3p, hsa-miR-92a-2-5p, hsa At least one miRNA from the following groups can also be preferred as target nucleic acids: hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-371a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR-1203, hsa-miR-122-5p, hsa-miR-4258, hsa-miR-4484, hsa-miR-4648, and hsa-miR-6780b-5p.

[0289] Among the above-mentioned miRNAs, for example, human genes containing the base sequences shown in any one of SEQ ID NO: 1 to 148, 466 to 478 (that is, hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR-150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, hsa-miR-6836-3p, hsa-miR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-miR-423-5p, hsa-miR-4454, hsa-miR-7109-5p, hsa-miR-6789-5p, hsa-miR-6877-5p, hsa-miR-4792, hsa-miR-4530, hsa-miR-7975, hsa-miR-6724-5p, hsa-miR-8073, hsa-miR-7977, hsa-miR-1231, hsa-miR-6799-5p, hsa-miR-615-5p, hsa-miR-4450, hsa-miR-6726-5p, hsa-miR-6875-5p, hsa-miR-4734, hsa-miR-16-5p, hsa-miR-602, hsa-miR-4651, hsa-miR-8069, hsa-miR-1238-5p, hsa-miR-6880-5p, hsa-miR-8072, hsa-miR-4723-5p, hsa-miR-4732-5p, hsa-miR-6125, hsa-miR-6090, hsa-miR-7114-5p, hsa-miR-564, hsa-miR-451a, hsa-miR-3135b, hsa-miR-4497, hsa-miR-4665-5p, hsa-miR-3622a-5p, hsa-miR-6850-5p, hsa-miR-6821-5p, hsa-miR-5100, hsa-miR-6872-3p, hsa-miR-4433-3p, hsa-miR-1227-5p, hsa-miR-3188, hsa-miR-7704, hsa-miR-3185, hsa-miR-1908-3p, hsa-miR-6781-5p, hsa-miR-6805-5p, hsa-miR-8089,hsa-miR-665, hsa-miR-4486, hsa-miR-6722-3p, hsa-miR-1260a, hsa-miR-4707-5p, hsa-miR-6741-5p, hsa-miR-1260b, hsa-miR-1246, hsa-miR-6845-5p, hsa-miR-4638-5p, hsa-miR-6085, hsa-miR-1228-3p, hsa-miR-4534, hsa-miR-5585-3p, hsa-miR-4741, hsa-miR-4433b-3p, hsa-miR-197-5 p, hsa-miR-718, hsa-miR-4513, hsa-miR-4446-3p, hsa-miR-619-5p, hsa-miR-6816-5p, hsa-miR-6778-5p, hsa-miR-24-3p, hsa-miR-1915-3p, hsa- miR-4665-3p, hsa-miR-4449, hsa-miR-6889-5p, hsa-miR-486-3p, hsa-miR-7113-3p, hsa-miR-642a-3p, hsa-miR-7847-3p, hsa-miR-6768-5p, hsa-miR-1290, hsa-miR-7108-5p, hsa-miR-92b-5p, hsa-miR-663b, hsa-miR-3940-5p, hsa-miR-4467, hsa-miR-6858-5p, hsa-miR-4417, hsa-miR-3665 hsa-miR-4736 hsa-miR-4687-3p hsa-miR-1908-5p hsa-miR-5195-3p hsa-miR-4286 hsa-miR-3679-3p hsa-miR-6791-5p hsa-miR-1202 hsa- miR-3656, hsa-miR-4746-3p, hsa-miR-3184-5p, hsa-miR-3937, hsa-miR-6515-3p, hsa-miR-6132, hsa-miR-187-5p, hsa-miR-7111-5p, hsa-miR-57 87, hsa-miR-6779-5p, hsa-miR-6808-5p, hsa-miR-6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa-miR-1233-5p, hsa-miR-328-5p, hsa-miR-4674hsa-miR-2110, hsa-miR-6076, hsa-miR-3619-3p, hsa-miR-92a-2-5p, hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-3 71a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR-1203, hsa-miR-122-5p, hsa-miR-4258, hsa-miR-4484, hsa-miR-4648, hsa- Genes, their families, transcripts, and mutants or derivatives thereof (including miR-6780b-5p, hsa-miR-4516, hsa-miR-4649-5p, hsa-miR-760, hsa-miR-3162-5p, hsa-miR-3178, hsa-miR-940, hsa-miR-4271, hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-3131, and hsa-miR-1343-3p), their families, their transcripts, and their mutants or derivatives. Here, genes, families, transcripts, mutants, and derivatives are as defined above.

[0290] The preferred target nucleic acid is a human gene containing the base sequence shown in any one of the sequence numbers 1 to 509, or its transcript, more preferably the transcript, i.e., miRNA, pri-miRNA or pre-miRNA as its precursor RNA.

[0291] The primary target genes are the hsa-miR-125a-3p gene, their homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0292] The second target genes are the hsa-miR-6893-5p gene, their homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0293] The third target gene is the hsa-miR-204-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0294] The fourth target gene is the hsa-miR-4476 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0295] The fifth target gene is the hsa-miR-4294 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0296] The sixth target gene is the hsa-miR-150-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0297] The seventh target gene is the hsa-miR-6729-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0298] The eighth target gene is the hsa-miR-7641 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0299] The ninth target gene is the hsa-miR-6765-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0300] The 10th target gene is the hsa-miR-6820-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0301] The 11th target gene is the hsa-miR-575 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0302] The 12th target gene is the hsa-miR-6836-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0303] The 13th target gene is the hsa-miR-1469 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0304] The 14th target gene is the hsa-miR-663a gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0305] The 15th target gene is the hsa-miR-6075 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0306] The 16th target gene is the hsa-miR-4634 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0307] The 17th target gene is the hsa-miR-423-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0308] The 18th target gene is the hsa-miR-4454 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0309] The 19th target gene is the hsa-miR-7109-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0310] The 20th target gene is the hsa-miR-6789-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0311] The 21st target gene is the hsa-miR-6877-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0312] The 22nd target gene is the hsa-miR-4792 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0313] The 23rd target gene is the hsa-miR-4530 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0314] The 24th target gene is the hsa-miR-7975 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0315] The 25th target gene is the hsa-miR-6724-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0316] The 26th target gene is the hsa-miR-8073 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0317] The 27th target gene is the hsa-miR-7977 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0318] The 28th target gene is the hsa-miR-1231 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0319] The 29th target gene is the hsa-miR-6799-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0320] The 30th target gene is the hsa-miR-615-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0321] The 31st target gene is the hsa-miR-4450 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0322] The 32nd target gene is the hsa-miR-6726-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0323] The 33rd target gene is the hsa-miR-6875-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0324] The 34th target gene is the hsa-miR-4734 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0325] The 35th target gene is the hsa-miR-16-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0326] The 36th target gene is the hsa-miR-602 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0327] The 37th target gene is the hsa-miR-4651 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0328] The 38th target gene is the hsa-miR-8069 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0329] The 39th target gene is the hsa-miR-1238-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0330] The 40th target gene is the hsa-miR-6880-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0331] The 41st target gene is the hsa-miR-8072 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0332] The 42nd target gene is the hsa-miR-4723-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0333] The 43rd target gene is the hsa-miR-4732-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0334] The 44th target gene is the hsa-miR-6125 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0335] The 45th target gene is the hsa-miR-6090 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0336] The 46th target gene is the hsa-miR-7114-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0337] The 47th target gene is the hsa-miR-564 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0338] The 48th target gene is the hsa-miR-451a gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0339] The 49th target gene is the hsa-miR-3135b gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0340] The 50th target gene is the hsa-miR-4497 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0341] The 51st target gene is the hsa-miR-4665-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0342] The 52nd target gene is the hsa-miR-3622a-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0343] The 53rd target gene is the hsa-miR-6850-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0344] The 54th target gene is the hsa-miR-6821-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0345] The 55th target gene is the hsa-miR-5100 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0346] The 56th target gene is the hsa-miR-6872-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0347] The 57th target gene is the hsa-miR-4433-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0348] The 58th target gene is the hsa-miR-1227-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0349] The 59th target gene is the hsa-miR-3188 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0350] The 60th target gene is the hsa-miR-7704 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0351] The 61st target gene is the hsa-miR-3185 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0352] The 62nd target gene is the hsa-miR-1908-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0353] The 63rd target gene is the hsa-miR-6781-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0354] The 64th target gene is the hsa-miR-6805-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0355] The 65th target gene is the hsa-miR-8089 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0356] The 66th target gene is the hsa-miR-665 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0357] The 67th target gene is the hsa-miR-4486 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0358] The 68th target gene is the hsa-miR-6722-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0359] The 69th target gene is the hsa-miR-1260a gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0360] The 70th target gene is the hsa-miR-4707-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0361] The 71st target gene is the hsa-miR-6741-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0362] The 72nd target gene is the hsa-miR-1260b gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0363] The 73rd target gene is the hsa-miR-1246 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0364] The 74th target gene is the hsa-miR-6845-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0365] The 75th target gene is the hsa-miR-4638-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0366] The 76th target gene is the hsa-miR-6085 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0367] The 77th target gene is the hsa-miR-1228-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0368] The 78th target gene is the hsa-miR-4534 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0369] The 79th target gene is the hsa-miR-5585-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0370] The 80th target gene is the hsa-miR-4741 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0371] The 81st target gene is the hsa-miR-4433b-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0372] The 82nd target gene is the hsa-miR-197-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0373] The 83rd target gene is the hsa-miR-718 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0374] The 84th target gene is the hsa-miR-4513 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0375] The 85th target gene is the hsa-miR-4446-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0376] The 86th target gene is the hsa-miR-619-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0377] The 87th target gene is the hsa-miR-6816-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0378] The 88th target gene is the hsa-miR-6778-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0379] The 89th target gene is the hsa-miR-24-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0380] The 90th target gene is the hsa-miR-1915-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0381] The 91st target gene is the hsa-miR-4665-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0382] The 92nd target gene is the hsa-miR-4449 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0383] The 93rd target gene is the hsa-miR-6889-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0384] The 94th target gene is the hsa-miR-486-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0385] The 95th target gene is the hsa-miR-7113-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0386] The 96th target gene is the hsa-miR-642a-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0387] The 97th target gene is the hsa-miR-7847-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0388] The 98th target gene is the hsa-miR-6768-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0389] The 99th target gene is the hsa-miR-1290 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0390] The 100th target gene is the hsa-miR-7108-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0391] The 101st target gene is the hsa-miR-92b-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0392] The 102nd target gene is the hsa-miR-663b gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0393] The 103rd target gene is the hsa-miR-3940-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0394] The 104th target gene is the hsa-miR-4467 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0395] The 105th target gene is the hsa-miR-6858-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0396] The 106th target gene is the hsa-miR-4417 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0397] The 107th target gene is the hsa-miR-3665 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0398] The 108th target gene is the hsa-miR-4736 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0399] The 109th target gene is the hsa-miR-4687-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0400] The 110th target gene is the hsa-miR-1908-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0401] The 111th target gene is the hsa-miR-5195-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0402] The 112th target gene is the hsa-miR-4286 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0403] The 113th target gene is the hsa-miR-3679-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0404] The 114th target gene is the hsa-miR-6791-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0405] The 115th target gene is the hsa-miR-1202 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0406] The 116th target gene is the hsa-miR-3656 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0407] The 117th target gene is the hsa-miR-4746-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0408] The 118th target gene is the hsa-miR-3184-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0409] The 119th target gene is the hsa-miR-3937 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0410] The 120th target gene is the hsa-miR-6515-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0411] The 121st target gene is the hsa-miR-6132 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0412] The 122nd target gene is the hsa-miR-187-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0413] The 123rd target gene is the hsa-miR-7111-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0414] The 124th target gene is the hsa-miR-5787 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0415] The 125th target gene is the hsa-miR-6779-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0416] The 126th target gene is the hsa-miR-6808-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0417] The 127th target gene is the hsa-miR-6774-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0418] The 128th target gene is the hsa-miR-4656 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0419] The 129th target gene is the hsa-miR-6806-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0420] The 130th target gene is the hsa-miR-1233-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0421] The 131st target gene is the hsa-miR-328-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0422] The 132nd target gene is the hsa-miR-4674 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0423] The 133rd target gene is the hsa-miR-2110 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0424] The 134th target gene is the hsa-miR-6076 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0425] The 135th target gene is the hsa-miR-3619-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0426] The 136th target gene is the hsa-miR-92a-2-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0427] The 137th target gene is the hsa-miR-128-1-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0428] The 138th target gene is the hsa-miR-638 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0429] The 139th target gene is the hsa-miR-2861 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0430] The 140th target gene is the hsa-miR-371a-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0431] The 141st target gene is the hsa-miR-211-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0432] The 142nd target gene is the hsa-miR-1273g-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0433] The 143rd target gene is the hsa-miR-1203 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0434] The 144th target gene is the hsa-miR-122-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0435] The 145th target gene is the hsa-miR-4258 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0436] The 146th target gene is the hsa-miR-4484 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0437] The 147th target gene is the hsa-miR-4648 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0438] The 148th target gene is the hsa-miR-6780b-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in the expression of this gene or its transcripts serving as biomarkers for biliary tract cancer.

[0439] The 149th target gene is the hsa-miR-4516 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0440] The 150th target gene is the hsa-miR-4649-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0441] The 151st target gene is the hsa-miR-760 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0442] The 152nd target gene is the hsa-miR-3162-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0443] The 153rd target gene is the hsa-miR-3178 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0444] The 154th target gene is the hsa-miR-940 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0445] The 155th target gene is the hsa-miR-4271 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0446] The 156th target gene is the hsa-miR-6769b-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0447] The 157th target gene is the hsa-miR-4508 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0448] The 158th target gene is the hsa-miR-6826-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0449] The 159th target gene is the hsa-miR-6757-5p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0450] The 160th target gene is the hsa-miR-3131 gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0451] The 161st target gene is the hsa-miR-1343-3p gene, its homologs, their transcripts, or their mutants or derivatives. To date, there are no reports of changes in gene or transcript expression serving as biomarkers for biliary tract cancer.

[0452] 2. Nucleic acid probes or primers for the detection of biliary tract cancer.

[0453] In this invention, nucleic acids that can specifically bind to the target nucleic acids described above as markers of biliary tract cancer can be used as nucleic acids for detecting or diagnosing biliary tract cancer, such as nucleic acid probes or primers.

[0454] In this invention, nucleic acid probes or primers capable of detecting or diagnosing biliary tract cancer can qualitatively and / or quantitatively determine the target nucleic acids used as biomarkers for biliary tract cancer, such as human-derived hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR-150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, hsa-miR-6836-3p, hsa -miR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-miR-423-5p, hsa-miR-4454, hsa-miR-7109-5p, hsa-miR-6789-5p, hsa-miR-6877-5p , hsa-miR-4792, hsa-miR-4530, hsa-miR-7975, hsa-miR-6724-5p, hsa-miR-8073, hsa-miR-7977, hsa-miR-1231, hsa-miR-6799-5p, hsa-miR-615-5 p, hsa-miR-4450, hsa-miR-6726-5p, hsa-miR-6875-5p, hsa-miR-4734, hsa-miR-16-5p, hsa-miR-602, hsa-miR-4651, hsa-miR-8069, hsa-miR-1238 -5p, hsa-miR-6880-5p, hsa-miR-8072, hsa-miR-4723-5p, hsa-miR-4732-5p, hsa-miR-6125, hsa-miR-6090, hsa-miR-7114-5p, hsa-miR-564, hsa-m iR-451a, hsa-miR-3135b, hsa-miR-4497, hsa-miR-4665-5p, hsa-miR-3622a-5p, hsa-miR-6850-5p, hsa-miR-6821-5p, hsa-miR-5100, hsa-miR-687 2-3p, hsa-miR-4433-3p, hsa-miR-1227-5p, hsa-miR-3188, hsa-miR-7704, hsa-miR-3185, hsa-miR-1908-3p, hsa-miR-6781-5p, hsa-miR-6805-5p,hsa-miR-8089, hsa-miR-665, hsa-miR-4486, hsa-miR-6722-3p, hsa-miR-1260a, hsa-miR-4707-5p, hsa-miR-6741-5p, hsa-miR-1260b, hsa-miR-12 46, hsa-miR-6845-5p, hsa-miR-4638-5p, hsa-miR-6085, hsa-miR-1228-3p, hsa-miR-4534, hsa-miR-5585-3p, hsa-miR-4741, hsa-miR-4433b-3p, h sa-miR-197-5p, hsa-miR-718, hsa-miR-4513, hsa-miR-4446-3p, hsa-miR-619-5p, hsa-miR-6816-5p, hsa-miR-6778-5p, hsa-miR-24-3p, hsa-miR- 1915-3p, hsa-miR-4665-3p, hsa-miR-4449, hsa-miR-6889-5p, hsa-miR-486-3p, hsa-miR-7113-3p, hsa-miR-642a-3p, hsa-miR-7847-3p, hsa-miR- 6768-5p, hsa-miR-1290, hsa-miR-7108-5p, hsa-miR-92b-5p, hsa-miR-663b, hsa-miR-3940-5p, hsa-miR-4467, hsa-miR-6858-5p, hsa-miR-4417, h sa-miR-3665, hsa-miR-4736, hsa-miR-4687-3p, hsa-miR-1908-5p, hsa-miR-5195-3p, hsa-miR-4286, hsa-miR-3679-3p, hsa-miR-6791-5p, hsa-miR-1202, hsa-miR-3656, hsa-miR-4746-3p, hsa-miR-3184-5p, hsa-miR-3937, hsa-miR-6515-3p, hsa-miR-6132, hsa-miR-187-5p, hsa-miR-7111-5p hsa-miR-5787 hsa-miR-6779-5p hsa-miR-4516 hsa-miR-4649-5p hsa-miR-760 hsa-miR-3162-5p hsa-miR-3178 hsa-miR-940 hsa-miR-4271hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-3131 or hsa-miR-1343-3p or combinations thereof, or their homologs, their transcripts, their mutants or derivatives, and hsa-miR-6808-5p, hsa-miR-6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa-miR-1233-5p, hsa-miR-328-5p, hsa-miR-4674, hsa-miR-2110, hsa- The following are considered to be present, expressed, or present in quantities of miR-6076, hsa-miR-3619-3p, hsa-miR-92a-2-5p, hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-371a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR-1203, hsa-miR-122-5p, hsa-miR-4258, hsa-miR-4484, hsa-miR-4648, or hsa-miR-6780b-5p, or combinations thereof, or their homologs, their transcripts, their mutants, or their derivatives.

[0455] Regarding the aforementioned target nucleic acids, compared to healthy individuals, their expression levels in subjects with biliary tract cancer can be either increased or decreased (hereinafter referred to as "increased / decreased"), depending on the type of target nucleic acid. Therefore, the nucleic acid of the present invention can measure the expression levels of the aforementioned target nucleic acids in bodily fluids derived from subjects suspected of having biliary tract cancer (e.g., humans) and bodily fluids derived from healthy individuals, and compare them, effectively for the detection of biliary tract cancer. Furthermore, the nucleic acid of the present invention can measure the expression levels of the aforementioned target nucleic acids in bodily fluids derived from subjects suspected of having biliary tract cancer (e.g., humans), as well as bodily fluids derived from patients with colorectal cancer, gastric cancer, esophageal cancer, liver cancer, and benign pancreatic and biliary tract diseases, and compare them, effectively for the specific detection of biliary tract cancer from other cancers, benign diseases, etc.

[0456] The nucleic acid probes or primers that can be used in this invention are nucleic acid probes that can specifically bind to polynucleotides composed of at least one of the base sequences shown in sequence numbers 1 to 125 (preferably sequence numbers 1, 2, 4 to 125) and 466 to 478, or primers for amplifying polynucleotides composed of at least one of the base sequences shown in sequence numbers 1 to 125 and 466 to 478.

[0457] The nucleic acid probes or primers that can be used in this invention may further include nucleic acid probes that can specifically bind to polynucleotides composed of at least one of the base sequences shown in sequence numbers 126 to 148, or primers for amplifying polynucleotides composed of at least one of the base sequences shown in sequence numbers 126 to 148.

[0458] Specifically, the aforementioned nucleic acid probes or primers comprise polynucleotide groups selected from the base sequences shown in any one of sequence numbers 1 to 509, or base sequences in which u is t, and their complementary polynucleotide groups; polynucleotide groups and their complementary polynucleotide groups that hybridize separately to DNA composed of base sequences complementary to the aforementioned base sequences under stringent conditions (described later); and combinations of one or more polynucleotides from polynucleotide groups comprising 15 or more, preferably 17 or more consecutive bases from the base sequences of these polynucleotide groups. These polynucleotides can be used as nucleic acid probes and primers for detecting the aforementioned biliary tract cancer markers as target nucleic acids.

[0459] More specifically, examples of nucleic acid probes or primers that can be used in this invention are one or more polynucleotides selected from the following polynucleotides (a) to (e).

[0460] (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0461] (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 1–125 and 466–478.

[0462] (c) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 1-125, 466-478, or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0463] (d) A polynucleotide comprising a base sequence complementary to the base sequence shown in any of the sequence numbers 1–125, 466–478, or a base sequence in which u is t, and,

[0464] (e) A polynucleotide that hybridizes with any of the polynucleotides (a) to (d) above under strict conditions.

[0465] The nucleic acid probes or primers that can be used in this invention may contain, in addition to at least one polynucleotide selected from the polynucleotides (a) to (e) above, polynucleotides selected from the polynucleotides (f) to (j) below.

[0466] (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0467] (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126 to 148.

[0468] (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequences 126 to 148 or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0469] (i) a polynucleotide comprising a base sequence complementary to the base sequence shown in any of sequences 126–148 or a base sequence in which u is t, and,

[0470] (j) A polynucleotide that hybridizes with any of the polynucleotides (f) to (i) above under strict conditions.

[0471] The term "fragment containing 15 or more consecutive bases" in the above-mentioned polynucleotides may include, for example, a number of bases in the polynucleotide sequence that is less than 15 consecutive bases, less than 17 consecutive bases, less than 19 consecutive bases, etc., but is not limited to this.

[0472] The polynucleotides or their fragments used in this invention can be either DNA or RNA.

[0473] The polynucleotides used in this invention can be produced using common techniques such as DNA recombination technology, PCR, and methods using an automated DNA / RNA synthesizer.

[0474] DNA recombination technology and PCR can use techniques described in, for example, Ausubel et al., Current Protocols in Molecular Biology, John Willey & Sons, US (1993); Sambrook et al., Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press, US (1989).

[0475] Human-derived hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR-150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, hsa-miR-6836-3p, hsa-miR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-miR-423-5p, hsa-miR-4454, hsa-miR-7109-5p, hsa-miR-6789-5p, hsa-miR-6877-5p, hsa-miR-4792, hsa-miR-4530, hsa-miR-7975, hsa-miR-6724-5p, hsa-miR-8073, hsa-miR-7977, hsa-miR-1231, hsa-miR-6799-5p, hsa-miR-615-5p, hsa-miR-4450, hsa-miR-6726-5p, hsa-miR-6875-5p, hsa-miR-4734, hsa-miR-16-5p, hsa-miR-602, hsa-miR-4651, hsa-miR-8069, hsa-miR-1238-5p, hsa-miR-6880-5p, hsa-miR-8072, hsa-miR-4723-5p, hsa-miR-4732-5p, hsa-miR-6125, hsa-miR-6090, hsa-miR-7114-5p, hsa-miR-564, hsa-miR-451a, hsa-miR-3135b, hsa-miR-4497, hsa-miR-4665-5p, hsa-miR-3622a-5p, hsa-miR-6850-5p, hsa-miR-6821-5p, hsa-miR-5100, hsa-miR-6872-3p, hsa-miR-4433-3p, hsa-miR-1227-5p, hsa-miR-3188, hsa-miR-7704, hsa-miR-3185, hsa-miR-1908-3p, hsa-miR-6781-5p, hsa-miR-6805-5p, hsa-miR-8089, hsa-miR-665, hsa-miR-4486, hsa-miR-6722-3p, as shown by serial numbers 1 to 148, 466 to 478hsa-miR-1260a, hsa-miR-4707-5p, hsa-miR-6741-5p, hsa-miR-1260b, hsa-miR-1246, hsa-miR-6845-5p, hsa-miR-4638-5p, hsa-miR-6085, hsa-miR-1228-3p, hsa-miR-4534, hsa-miR-5585-3p, hsa-miR-4741, hsa-miR-4433b-3p, hsa-miR-197-5p, hsa-miR-718, hsa-miR-4513, hsa-miR-4446-3 p, hsa-miR-619-5p, hsa-miR-6816-5p, hsa-miR-6778-5p, hsa-miR-24-3p, hsa-miR-1915-3p, hsa-miR-4665-3p, hsa-miR-4449, hsa-miR-6889-5p hsa-miR-486-3p, hsa-miR-7113-3p, hsa-miR-642a-3p, hsa-miR-7847-3p, hsa-miR-6768-5p, hsa-miR-1290, hsa-miR-7108-5p, hsa-miR-92b-5p, hsa-miR-663b, hsa-miR-3940-5p, hsa-miR-4467, hsa-miR-6858-5p, hsa-miR-4417, hsa-miR-3665, hsa-miR-4736, hsa-miR-4687-3p, hsa-miR-1908 -5p, hsa-miR-5195-3p, hsa-miR-4286, hsa-miR-3679-3p, hsa-miR-6791-5p, hsa-miR-1202, hsa-miR-3656, hsa-miR-4746-3p, hsa-miR-3184-5p, h sa-miR-3937, hsa-miR-6515-3p, hsa-miR-6132, hsa-miR-187-5p, hsa-miR-7111-5p, hsa-miR-5787, hsa-miR-6779-5p, hsa-miR-6808-5p, hsa-miR -6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa-miR-1233-5p, hsa-miR-328-5p, hsa-miR-4674, hsa-miR-2110, hsa-miR-6076, hsa-miR-3619-3phsa-miR-92a-2-5p, hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-371a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR-1203, hsa-miR-122-5p, hsa-miR-4258, hsa-miR-4484, hsa-miR-4648, and hsa-miR-6780b-5p are known, and their methods of acquisition are also known, as described above. Therefore, by cloning this gene, polynucleotides that can be used as nucleic acid probes or primers in this invention can be prepared.

[0476] Such nucleic acid probes or primers can be chemically synthesized using automated DNA synthesis devices. This synthesis typically employs the phosphoramidite method, which can automatically synthesize single-stranded DNA up to approximately 100 base pairs. Automated DNA synthesis devices are commercially available from companies such as Polygen, ABI, and Applied BioSystems.

[0477] Alternatively, the polynucleotides of the present invention can also be prepared by cDNA cloning. cDNA cloning technology can utilize, for example, the Wako microRNA Cloning Kit.

[0478] Here, the sequences of nucleic acid probes and primers used to detect polynucleotides composed of the base sequences shown in any of the sequences 1–148 and 466–478 are not present in the organism as miRNAs or their precursors. For example, the base sequences shown in sequences 51 and 91 are generated from the precursor shown in sequence 201, but this precursor has… Figure 1 The hairpin-like structure shown has mismatched base sequences in sequences 51 and 91. Therefore, base sequences that are perfectly complementary to those in sequence 51 or 91 are not naturally generated in living organisms. Similarly, the nucleic acid probes and primers used to detect the base sequences shown in sequences 1–148 and 466–478 have artificial base sequences not present in living organisms.

[0479] 3. Kits or devices for detecting bile duct cancer

[0480] The present invention also provides a kit or device for detecting biliary tract cancer comprising one or more target nucleic acids used as markers of biliary tract cancer, which can be used as nucleic acid probes or primers in the present invention (which may include mutants, fragments or derivatives; hereinafter sometimes referred to as detection polynucleotides).

[0481] In this invention, the target nucleic acids used as biomarkers for biliary tract cancer are preferably selected from group 1 of the following:

[0482] miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6836 -3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, mi R-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4 651, miR-8069, miR-1238-5p, miR-6880-5p, miR-8072, miR-4723-5p, miR-4732-5p, miR-6125, miR-6090, miR-7114-5p, miR-564, miR-451a, miR-3135 b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3p, miR-1227-5p, miR-3188, miR-7704, mi R-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a, miR-4707-5p, miR-6741-5p, miR-1260b miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-4433b-3p, miR-197-5p, miR-718, miR-4513 miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449, miR-6889-5p, miR-486-3p, miR-7113-3pmiR-642a-3p,miR-7847-3p,miR-6768-5p,miR-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR-4467,miR-6858-5p,miR- 4417,miR-3665,miR-4736,miR-4687-3p,miR-1908-5p,miR-5195-3p,miR-4286,miR-3679-3p,miR-6791-5p,miR-1202,miR-3656,miR- 4746-3p,miR-3184-5p,miR-3937,miR-6515-3p,miR-6132,miR-187-5p,miR-7111-5p,miR-5787,miR-6779-5p,miR-4516,miR-4649-5p ,miR-760,miR-3162-5p,miR-3178,miR-940,miR-4271,miR-6769b-5p,miR-4508,miR-6826-5p,miR-6757-5p,miR-3131 and miR-1343-3p. ,

[0483] Depending on the circumstances, the additional target nucleic acids that can be used for assay are preferably selected from the following group 2: miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, and miR-6780b-5p.

[0484] The kit or device of the present invention contains a nucleic acid capable of specifically binding to the target nucleic acid described above as a marker of biliary tract cancer, preferably containing the nucleic acid probe or primer described in section 2 above, and specifically containing one or more polynucleotides selected from the polynucleotide classes described in section 2 above, or mutants of such polynucleotides.

[0485] Specifically, the kit or device of the present invention may contain at least one: a polynucleotide containing a base sequence represented by any one of the sequence numbers 1 to 125 and 466 to 478 or a base sequence in which u is t (or composed of a base sequence represented by any one of the sequence numbers 1 to 125 and 466 to 478 or a base sequence in which u is t), a polynucleotide containing its complementary sequence (or composed of its complementary sequence), a polynucleotide hybridizing with these polynucleotides under stringent conditions, or a mutant or fragment of these polynucleotide sequences containing 15 or more consecutive bases.

[0486] The kit or device of the present invention may further comprise one or more of the following: a polynucleotide comprising the base sequence shown in any of the serial numbers 126 to 148 or a base sequence in which u is t (or composed of the base sequence shown in any of the serial numbers 126 to 148 or a base sequence in which u is t), a polynucleotide comprising its complementary sequence (or composed of its complementary sequence), a polynucleotide hybridizing with these polynucleotides under stringent conditions, or a mutant or fragment of these polynucleotide sequences comprising 15 or more consecutive bases.

[0487] The kit or device of the present invention may contain fragments selected from, for example, one or more, preferably two or more, polynucleotides selected from (1) to (2) below.

[0488] (1) A polynucleotide containing 15 or more consecutive bases in the base sequence of any of the sequence numbers 1 to 125 and 466 to 478, where u is t, or the complementary sequence thereof.

[0489] (2) A polynucleotide containing 15 or more consecutive bases in the base sequence of any of the sequences shown in sequence numbers 126 to 148, where u is t, or in the complementary sequence thereof.

[0490] In a preferred embodiment, the polynucleotide is a polynucleotide composed of a base sequence represented by any one of sequence numbers 1 to 125, 466 to 478 or a base sequence in which u is t; a polynucleotide composed of its complementary sequence; a polynucleotide that hybridizes with these polynucleotides under stringent conditions; or a mutant of these polynucleotides containing 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases.

[0491] Furthermore, in a preferred embodiment, the polynucleotide is a polynucleotide composed of a base sequence shown in any of the sequences 126 to 148 or a base sequence in which u is t; a polynucleotide composed of its complementary sequence; a polynucleotide that hybridizes with these polynucleotides under stringent conditions; or a mutant of these polynucleotides containing 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases.

[0492] In a preferred embodiment, the above-mentioned fragment may be a polynucleotide containing 15 or more, preferably 17 or more, and more preferably 19 or more consecutive bases.

[0493] In this invention, the size of the polynucleotide fragment is, for example, within the range of 15 to less than the total number of bases in the sequence of each polynucleotide, 17 to less than the total number of bases in the sequence, 19 to less than the total number of bases in the sequence, etc.

[0494] Specifically, examples of the above-described polynucleotide combinations constituting the kits or devices of the present invention include any combination of the above-described polynucleotides consisting of the base sequences represented by the sequence numbers shown in Table 1 below (sequence numbers 1-148 and 466-478 corresponding to the miRNA markers in Table 1) or their complementary sequences. These are strictly illustrative, and all other possible combinations are included in the present invention.

[0495] For example, in the present invention, as the above-described combination constituting a kit or device for distinguishing between biliary tract cancer and healthy individuals, it is desirable that the above-described polynucleotides composed of the base sequences shown by the serial numbers shown in Table 1 are combined in two or more combinations. Generally, a combination of two can achieve sufficient performance.

[0496] Specifically, as a combination of two polynucleotides consisting of a base sequence used to distinguish between bile duct cancer and a healthy body or its complementary sequence, the combination of two of the above-mentioned polynucleotides selected from the base sequences shown in sequence numbers 1 to 148 and 466 to 478 preferably includes at least one newly discovered polynucleotide combination consisting of a base sequence shown in sequence numbers 1 to 125 and 466 to 478.

[0497] Furthermore, as a combination of cancer-type-specific polynucleotides that can distinguish not only healthy individuals from biliary tract cancer but also healthy individuals from other cancers, it is preferably, for example, a combination of at least one polynucleotide selected from a group consisting of polynucleotides composed of the base sequences shown in sequence numbers 1, 4, 5, 11, 12, 15, 23, 29, 39, 40, 54, 76, 79, 91, 103, 115, 121, 134, 143, 466, 469, 472, 473 and 474 or their complementary sequences (hereinafter, this group is referred to as "cancer-type-specific polynucleotide group 1") and a plurality of polynucleotides with other sequence numbers.

[0498] Furthermore, as a combination of cancer-type-specific polynucleotides that can distinguish not only healthy individuals from biliary tract cancer but also healthy individuals from other cancers, it is more preferably a combination of multiple polynucleotides selected from cancer-type-specific polynucleotide group 1.

[0499] Furthermore, as a combination of cancer-type-specific polynucleotides that can distinguish not only healthy individuals from biliary tract cancer but also healthy individuals from other cancers, among the combinations of multiple polynucleotides selected from cancer-type-specific polynucleotide group 1, it is more preferable to include a combination of polynucleotides selected from the group consisting of polynucleotides composed of the base sequences shown in sequence numbers 4, 5, 12, 15 and 40 or their complementary sequences contained in cancer-type-specific polynucleotide group 1 (hereinafter, this group is referred to as "cancer-type-specific polynucleotide group 2").

[0500] The number of the aforementioned combinations of cancer-type specific polynucleotides can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more, more preferably 4 or more. Generally, a combination of 4 can achieve sufficient performance.

[0501] Hereinafter, examples are given, without limitation, of a polynucleotide consisting of the base sequence shown in Serial No. 4 or its complementary sequence, and a polynucleotide consisting of the base sequence shown in Serial No. 4 of three polynucleotides selected from cancer type-specific polynucleotide group 1 or its complementary sequence.

[0502] (1) Combinations of serial numbers 4, 15, 54, and 115 (markers: miR-4476, miR-6075, miR-6821-5p, miR-1202)

[0503] (2) Combinations of serial numbers 4, 5, 12, 76 (markers: miR-4476, miR-4294, miR-6836-3p, miR-6085)

[0504] (3) Combinations of serial numbers 4, 5, 12, and 115 (markers: miR-4476, miR-4294, miR-6836-3p, miR-1202)

[0505] (4) Combinations of serial numbers 4, 12, 15, 474 (markers: miR-4476, miR-6836-3p, miR-6075, miR-4508)

[0506] (5) Combinations of serial numbers 4, 15, 29, and 115 (markers: miR-4476, miR-6075, miR-6799-5p, miR-1202)

[0507] Hereinafter, examples are given, without limitation, of a polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence, and a polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence of three polynucleotides selected from cancer type-specific polynucleotide group 1.

[0508] (1) Combinations of serial numbers 5, 76, 12, 115 (markers: hsa-miR-4294, hsa-miR-6085, hsa-miR-6836-3p, hsa-miR-1202)

[0509] (2) Combinations of serial numbers 5, 76, 54, 115 (markers: hsa-miR-4294, hsa-miR-6085, hsa-miR-6821-5p, hsa-miR-1202)

[0510] (3) Combinations of serial numbers 5, 23, 12, 115 (markers: hsa-miR-4294, hsa-miR-4530, hsa-miR-6836-3p, hsa-miR-1202)

[0511] (4) Combinations of serial numbers 5, 12, 115, 91 (markers: hsa-miR-4294, hsa-miR-6836-3p, hsa-miR-1202, hsa-miR-4665-3p)

[0512] (5) Combinations of serial numbers 5, 1, 23, 4 (markers: hsa-miR-4294, hsa-miR-125a-3p, hsa-miR-4530, hsa-miR-4476)

[0513] Hereinafter, combinations of polynucleotides consisting of the base sequence shown in sequence number 12 or its complementary sequence, and polynucleotides consisting of the base sequence shown in sequence number of three polynucleotides selected from cancer type-specific polynucleotide group 1, or their complementary sequences, are illustrated without limitation.

[0514] (1) Combinations of serial numbers 5, 12, 29, 115 (markers: miR-4294, miR-6836-3p, miR-6799-5p, miR-1202)

[0515] (2) Combinations of serial numbers 12, 15, 23, and 115 (markers: miR-6836-3p, miR-6075, miR-4530, miR-1202)

[0516] (3) Combinations of serial numbers 5, 12, 115, 469 (markers: miR-4294, miR-6836-3p, miR-3162-5p, miR-1202)

[0517] (4) Combinations of serial numbers 5, 12, 115, 472 (markers: miR-4294, miR-6836-3p, miR-1202, miR-4271)

[0518] (5) Combinations of serial numbers 5, 12, 76, and 115 (markers: miR-4294, miR-6085, miR-1202, miR-6836-3p)

[0519] Hereinafter, combinations of polynucleotides consisting of the base sequence shown in sequence number 15 or its complementary sequence, and polynucleotides consisting of the base sequence shown in sequence number of three polynucleotides selected from cancer type-specific polynucleotide group 1, are illustrated without limitation.

[0520] (1) Combinations of serial numbers 15, 29, 1, 12 (markers: hsa-miR-6075, hsa-miR-6799-5p, hsa-miR-125a-3p, hsa-miR-6836-3p)

[0521] (2) Combinations of serial numbers 15, 12, 11, 143 (markers: hsa-miR-6075, hsa-miR-6836-3p, hsa-miR-575, hsa-miR-1203)

[0522] (3) Combinations of serial numbers 15, 76, 121, 39 (markers: hsa-miR-6075, hsa-miR-6085, hsa-miR-6132, hsa-miR-1238-5p)

[0523] (4) Combinations of serial numbers 15, 76, 54, 121 (markers: hsa-miR-6075, hsa-miR-6085, hsa-miR-6821-5p, hsa-miR-6132)

[0524] (5) Combinations of serial numbers 15, 40, 1, 23 (markers: hsa-miR-6075, hsa-miR-6880-5p, hsa-miR-125a-3p, hsa-miR-4530)

[0525] Hereinafter, examples are given, without limitation, of a polynucleotide consisting of the base sequence shown in sequence number 40 or its complementary sequence, and a polynucleotide consisting of the base sequence shown in sequence number of three polynucleotides selected from cancer type-specific polynucleotide group 1 or its complementary sequence.

[0526] (1) Combinations of serial numbers 12, 40, 472, and 473 (markers: miR-6836-3p, miR-6880-5p, miR-4271, and miR-6769b-5p).

[0527] (2) Combinations of serial numbers 12, 23, 40, 466 (markers: miR-6836-3p, miR-4530, miR-6880-5p, miR-4516)

[0528] (3) Combinations of serial numbers 12, 23, 40, and 134 (markers: miR-6836-3p, miR-4530, miR-6880-5p, miR-6076)

[0529] (4) Combinations of serial numbers 15, 40, 121, and 134 (markers: miR-6075, miR-6880-5p, miR-6132, miR-6076)

[0530] (5) Combinations of serial numbers 15, 40, 54, and 76 (markers: miR-6075, miR-6880-5p, miR-6821-5p, miR-6085)

[0531] In addition to the polynucleotides described above (which may include mutants, fragments, or derivatives), the kits or devices of the present invention may also contain known or future-discovered polynucleotides capable of detecting biliary tract cancer.

[0532] In addition to the polynucleotides described above, the kit of the present invention may also contain antibodies for the determination of known biomarkers for biliary tract cancer detection, such as CEA, CA19-9, SPan-1, DUPAN-2, CA50, CA195, IL-6, CA242, TAG-72, urinary fucose, POA, and TPS.

[0533] The polynucleotides contained in the kit of the present invention can be packaged individually or in arbitrary combinations in different containers.

[0534] The kit of the present invention may also include a kit for extracting nucleic acids (e.g., total RNA) from body fluids, cells or tissues, a fluorescent labeling substance, an enzyme and culture medium for nucleic acid amplification, and an instruction manual.

[0535] The apparatus of the present invention is for measuring cancer biomarkers, wherein the polynucleotides and other nucleic acids described above are used to measure cancer biomarkers bound to or attached to a solid phase. Examples of solid phase materials include plastic, paper, glass, and silicon; plastic is preferred for ease of processing. The shape of the solid phase is arbitrary, for example, square, circular, rectangular, or membrane-like. The apparatus of the present invention includes, for example, devices for measurement using hybridization techniques; specifically, examples include blotting devices, nucleic acid arrays (e.g., microarrays, DNA chips, RNA chips, etc.).

[0536] Nucleic acid array technology is a technique that uses methods to create arrays such as chips by binding or attaching nucleic acids one by one, and then uses these arrays to determine target nucleic acids by hybridization. These methods include: spotting nucleic acids onto the surface of a solid phase that has undergone surface treatments such as L-lysine coating and / or the introduction of functional groups such as amino and carboxyl groups using a high-density dispensing machine called a spotter or arrayer; spraying nucleic acids onto the solid phase using an inkjet printer that ejects tiny droplets through a nozzle via a piezoelectric element; and sequentially synthesizing nucleotides on the solid phase.

[0537] The kit or device of the present invention comprises nucleic acids capable of specifically binding to at least one, preferably at least two, more preferably at least three, and most preferably at least five to all polynucleotides of the miRNAs of group 1 described above as markers of biliary tract cancer. The kit or device of the present invention may further comprise, depending on the circumstances, nucleic acids capable of specifically binding to at least one, preferably at least two, more preferably at least three, and most preferably all five polynucleotides of the miRNAs of group 2 described above as markers of biliary tract cancer.

[0538] The kit or device of the present invention can be used for the detection of biliary tract cancer as described in section 4.

[0539] 4. Detection methods for bile duct cancer

[0540] The present invention further provides a method for detecting biliary tract cancer, comprising: using the kit or device of the present invention described in section 3 above (containing the above-described nucleic acids that can be used in the present invention), to in vitro determine the presence of the following groups in a sample: miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6836-3p, miR-1469, miR-663a, miR-6075, miR-4634, and miR-423-5p. ,miR-4454,miR-7109-5p,miR-6789-5p,miR-6877-5p,miR-4792,miR-4530,miR-7975,miR-6724-5p,miR-8073,miR-7977,miR-1231,miR-6799-5p, miR-615-5p,miR-4450,miR-6726-5p,miR-6875-5p,miR-4734,miR-16-5p,miR-602,miR-4651,miR-8069,miR-1238-5p,miR-6880-5p,miR-8072,miR -4723-5p,miR-4732-5p,miR-6125,miR-6090,miR-7114-5p,miR-564,miR-451a,miR-3135b,miR-4497,miR-4665-5p,miR-3622a-5p,miR-6850-5p, miR-6821-5p,miR-5100,miR-6872-3p,miR-4433-3p,miR-1227-5p,miR-3188,miR-7704,miR-3185,miR-1908-3p,miR-6781-5p,miR-6805-5p,miR-8 089,miR-665,miR-4486,miR-6722-3p,miR-1260a,miR-4707-5p,miR-6741-5p,miR-1260b,miR-1246,miR-6845-5p,miR-4638-5p,miR-6085,miR-1 228-3p,miR-4534,miR-5585-3p,miR-4741,miR-4433b-3p,miR-197-5p,miR-718,miR-4513,miR-4446-3p,miR-619-5p,miR-6816-5p,miR-6778-5p,miR-24-3p,miR-1915-3p,miR-4665-3p,miR-4449,miR-6889-5p,miR-486-3p,miR-7113-3p,miR-642a-3p,miR-7847-3p,miR-6768-5p,miR-12 90.miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR-4467,miR-6858-5p,miR-4417,miR-3665,miR-4736,miR-4687-3p,miR-1908-5p,m The expression levels of bile duct cancer-derived genes from miR-5195-3p, miR-4286, miR-3679-3p, miR-6791-5p, miR-1202, miR-3656, miR-4746-3p, miR-3184-5p, miR-3937, miR-6515-3p, miR-6132, miR-187-5p, miR-7111-5p, miR-5787, and miR-6779-5p, and, depending on the circumstances, selected from the following groups: miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, and miR-1233-5p. miR-328-5p,miR-4674,miR-2110,miR-6076,miR-3619-3p,miR-92a-2-5p,miR-128-1-5p,miR-638,miR-2861,miR-371a-5p,miR-211-3p,miR- 1273g-3p,miR-1203,miR-122-5p,miR-4258,miR-4484,miR-4648,miR-6780b-5p,miR-4516,miR-4649-5p,miR-760,miR-3162-5p,miR-3178,m The expression levels of genes derived from biliary tract cancer, including iR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p, and the expression levels of one or more of the biliary tract cancer-derived genes shown, are further evaluated using blood, serum, plasma, or other samples collected from subjects suspected of having biliary tract cancer and healthy individuals (including those without biliary tract cancer). The expression levels of the aforementioned genes in the samples are compared with the control expression levels in healthy individuals. For example, if the two expression levels are compared, and there is a statistically significant difference in the expression levels of the target nucleic acids in the samples, the subject is evaluated as having biliary tract cancer.

[0541] The method described above in this invention enables low-invasive early diagnosis of cancer with high sensitivity and specificity, thereby leading to early treatment and improved prognosis. Furthermore, it can monitor disease progression and the effectiveness of surgical treatment, radiotherapy, and chemotherapy.

[0542] For the method of extracting genes from samples such as blood, serum, and plasma according to the present invention, the RNA extraction reagent from the 3D-Gene (registered trademark) RNA extraction reagent from liquid sample kit (Toray Industries, Inc.) is particularly preferred. However, the general acidic phenol method (acid guanidinium-phenol-chloroform (AGPC) method) can also be used, as can Trizol (registered trademark) (Life Technologies Inc.), or RNA extraction reagents containing acidic phenol such as Trizol (Life Technologies Inc.) or Isogen (Nippon Gene Inc.) can be added and prepared. Furthermore, kits such as the miRNeasy (registered trademark) Mini Kit (Qiagen Inc.) can be used, but the method is not limited to these methods.

[0543] The present invention further provides the application of the kit or apparatus of the present invention for the in vitro detection of expression products of miRNA genes derived from biliary tract cancer in specimens of test subject origin.

[0544] In the methods described above, the kits or devices used above comprise polynucleotides that can be used in this invention, either individually or in all possible combinations, as described above.

[0545] In the detection or (genetic) diagnosis of biliary tract cancer of the present invention, the polynucleotides contained in the kit or device of the present invention can be used as probes or primers. When used as primers, TaqMan MicroRNA Assays from Life Technologies, the miScript PCR System from Qiagen, etc., can be used, but are not limited to these methods.

[0546] The polynucleotides contained in the kit or device of the present invention can be used as primers or probes in conventional methods for the specific detection of specific genes, such as hybridization techniques like RNA blotting, DNA blotting, in situ hybridization, Northern hybridization, and Southern hybridization, and quantitative amplification techniques like quantitative RT-PCR. As the test sample, depending on the type of detection method used, bodily fluids such as blood, serum, plasma, and urine are collected from the subject. Alternatively, total RNA modulated from such bodily fluids using the above methods can be used, and further, various polynucleotides containing cDNA modulated based on this RNA can be used.

[0547] The kit or device of the present invention is useful for the diagnosis of biliary tract cancer or the detection of its presence or absence. Specifically, the detection of biliary tract cancer using the kit or device can be performed as follows: From a subject suspected of having biliary tract cancer, using samples such as blood, serum, plasma, or urine, the expression level of genes detected by the nucleic acid probes or primers contained in the kit or device is detected in vitro. If the expression level of the target miRNA marker, determined by measuring at least one base sequence or its complementary sequence represented by sequence numbers 1-125, 466-478, and, depending on the case, at least one base sequence or its complementary sequence represented by sequence numbers 126-148 in the blood, serum, plasma, or urine samples of a subject suspected of having biliary tract cancer, is statistically significantly different from their expression levels in the blood, serum, plasma, or urine samples of a healthy individual, the subject can be evaluated as having biliary tract cancer.

[0548] The method of this invention can be combined with imaging diagnostic methods such as abdominal ultrasound, CT scan, endoscopic retrograde cholangiopancreatography (ERCP), and endoscopic ultrasound. This method can specifically detect biliary tract cancer and can substantially distinguish it from other cancers. Particularly in the case of pancreatic cancer, miRNA markers shared with some biliary tract cancers can be used; however, biliary tract cancer and pancreatic cancer can be distinguished by using discriminative discrimination boundaries, or by combining it with other diagnostic methods such as the aforementioned imaging diagnostic methods.

[0549] The detection method using the kit or device of the present invention to detect whether a sample contains expression products of genes derived from biliary tract cancer or not includes expression products of genes derived from biliary tract cancer includes: collecting bodily fluids such as blood, serum, plasma, and urine from a subject, and using one or more polynucleotides (including mutants, fragments, or derivatives) selected from the polynucleotide group of the present invention to determine the expression level of the target gene contained therein, thereby evaluating the presence or detection of biliary tract cancer. Furthermore, the biliary tract cancer detection method of the present invention can also be used, for example, in patients with biliary tract cancer, to evaluate or diagnose whether the disease has improved or the degree of improvement after administration of therapeutic drugs for improving the disease.

[0550] The method of the present invention may include, for example, the following steps (a), (b), and (c):

[0551] (a) The step of contacting a test sample derived from the test subject in vitro with the polynucleotides in the kit or device of the present invention.

[0552] (b) The step of using the above-mentioned polynucleotides as nucleic acid probes or primers to determine the expression level of target nucleic acid in the sample.

[0553] (c) Based on the results of (b), the steps to evaluate the presence or absence of biliary carcinoma (cells) in the subject.

[0554] Specifically, the present invention provides a method for detecting biliary tract cancer, which includes: using a substance capable of binding to a selected miRNA from miR-125a-3p, miR-6893-5p, miR-204-3p, miR-4476, miR-4294, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-6836-3p, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4651, miR-8069, miR-1238-5p, miR-6880-5p, miR-8072, miR-4723-5p, miR-4732-5p, miR-6125, miR-6090, miR-7114-5p, miR-564, miR-451a, miR-3135b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3p, miR-1227-5p, miR-3188, miR-7704, miR-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a, miR-4707-5p, miR-6741-5p, miR-1260b, miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-4433b-3p, miR-197-5p, miR-718, miR-4513, miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449miR-6889-5p,miR-486-3p,miR-7113-3p,miR-642a-3p,miR-7847-3p,miR-6768-5p,mi R-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR-4467,miR-6858-5p,m iR-4417,miR-3665,miR-4736,miR-4687-3p,miR-1908-5p,miR-5195-3p,miR-4286,mi R-3679-3p,miR-6791-5p,miR-1202,miR-3656,miR-4746-3p,miR-3184-5p,miR-3937,m The expression level of the target nucleic acid in the sample of the test subject is determined by specifically binding to at least one, preferably at least two, of the following polynucleotides: iR-6515-3p, miR-6132, miR-187-5p, miR-7111-5p, miR-5787, miR-6779-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p. The measured expression level and the measured control expression level of healthy subjects are used to evaluate in vitro whether the test subject has biliary tract cancer or not. ,

[0555] The term "evaluation" in this instruction manual does not refer to a physician's judgment, but rather to an evaluation and support based on the results of in vitro examinations.

[0556] As described above, in a preferred embodiment of the method of the present invention, specifically, miR-125a-3p is hsa-miR-125a-3p, miR-6893-5p is hsa-miR-6893-5p, miR-204-3p is hsa-miR-204-3p, miR-4476 is hsa-miR-4476, miR-4294 is hsa-miR-4294, miR-150-3p is hsa-miR-150-3p, miR-6729-5p is hsa-miR-6729-5p, miR-7641 is hsa-miR-7641, and miR-6765-3p is hsa- miR-6765-3p, miR-6820-5p is hsa-miR-6820-5p, miR-575 is hsa-miR-575, miR-6836-3p is hsa-miR-6836-3p, miR-1469 is hsa-miR-1469, miR-663a is hsa-miR-663a, miR-6075 is hsa-miR-6075, miR-4634 is hsa-miR-4634, miR-423-5p is hsa-miR-423-5p, miR-4454 is hsa-miR-4454, miR-7109-5p is hsa-miR-71 09-5p, miR-6789-5p is hsa-miR-6789-5p, miR-6877-5p is hsa-miR-6877-5p, miR-4792 is hsa-miR-4792, miR-4530 is hsa-miR-4530, miR-7975 is hsa-miR-7975, miR-6724-5p is hsa-miR-6724-5p, miR-8073 is hsa-miR-8073, miR-7977 is hsa-miR-7977, miR-1231 is hsa-miR-1231, miR-6799-5p is hsa-miR-6799 -5p, miR-615-5p is hsa-miR-615-5p, miR-4450 is hsa-miR-4450, miR-6726-5p is hsa-miR-6726-5p, miR-6875-5p is hsa-miR-6875-5p, miR-4734 is hsa-miR-4734, miR-16-5p is hsa-miR-16-5p, miR-602 is hsa-miR-602, miR-4651 is hsa-miR-4651, miR-8069 is hsa-miR-8069, miR-1238-5p is hsa-miR-1238-5p.miR-6880-5p is hsa-miR-6880-5p, miR-8072 is hsa-miR-8072, miR-4723-5p is hsa-miR-4723-5p, miR-4732-5p is hsa-miR-4732-5p, miR-6125 is hsa-miR-6125, miR-6090 is hsa-miR-6090, miR-7114-5p is hsa-miR-7114-5p, miR-564 is hsa-miR-564, miR-451a is hsa-miR-451a, miR-3135b is hsa-miR-3135b, m iR-4497 is hsa-miR-4497, miR-4665-5p is hsa-miR-4665-5p, miR-3622a-5p is hsa-miR-3622a-5p, miR-6850-5p is hsa-miR-6850-5p, miR-6821-5p is hsa-miR-6821-5p, miR-5100 is hsa-miR-5100, miR-6872-3p is hsa-miR-6872-3p, miR-4433-3p is hsa-miR-4433-3p, miR-1227-5p is hsa-miR-1227-5p, miR-3 188 is hsa-miR-3188, miR-7704 is hsa-miR-7704, miR-3185 is hsa-miR-3185, miR-1908-3p is hsa-miR-1908-3p, miR-6781-5p is hsa-miR-6781-5p, miR-6805-5p is hsa-miR-6805-5p, miR-8089 is hsa-miR-8089, miR-665 is hsa-miR-665, miR-4486 is hsa-miR-4486, miR-6722-3p is hsa-miR-6722-3p, miR-1260 a is hsa-miR-1260a, miR-4707-5p is hsa-miR-4707-5p, miR-6741-5p is hsa-miR-6741-5p, miR-1260b is hsa-miR-1260b, miR-1246 is hsa-miR-1246, miR-6845-5p is hsa-miR-6845-5p, miR-4638-5p is hsa-miR-4638-5p, miR-6085 is hsa-miR-6085, miR-1228-3p is hsa-miR-1228-3p, and miR-4534 is hsa-miR-4534.miR-5585-3p is hsa-miR-5585-3p, miR-4741 is hsa-miR-4741, miR-4433b-3p is hsa-miR-4433b-3p, miR-197-5p is hsa-miR-197-5p, miR-718 is hsa-miR-718, miR-4513 is hsa-miR-4513, miR-4446-3p is hsa-miR-4446-3p, miR-619-5p is hsa-miR-619-5p, miR-6816-5p is hsa-miR-6816-5p, miR-6778-5p is h sa-miR-6778-5p, miR-24-3p is hsa-miR-24-3p, miR-1915-3p is hsa-miR-1915-3p, miR-4665-3p is hsa-miR-4665-3p, miR-4449 is hsa-miR-4449, miR-6889-5p is hsa-miR-6889-5p, miR-486-3p is hsa-miR-486-3p, miR-7113-3p is hsa-miR-7113-3p, miR-642a-3p is hsa-miR-642a-3p, miR-7847-3p is hsa-m iR-7847-3p, miR-6768-5p is hsa-miR-6768-5p, miR-1290 is hsa-miR-1290, miR-7108-5p is hsa-miR-7108-5p, miR-92b-5p is hsa-miR-92b-5p, miR-663b is hsa-miR-663b, miR-3940-5p is hsa-miR-3940-5p, miR-4467 is hsa-miR-4467, miR-6858-5p is hsa-miR-6858-5p, miR-4417 is hsa-miR-4417, miR-36 65 is hsa-miR-3665, miR-4736 is hsa-miR-4736, miR-4687-3p is hsa-miR-4687-3p, miR-1908-5p is hsa-miR-1908-5p, miR-5195-3p is hsa-miR-5195-3p, miR-4286 is hsa-miR-4286, miR-3679-3p is hsa-miR-3679-3p, miR-6791-5p is hsa-miR-6791-5p, miR-1202 is hsa-miR-1202, and miR-3656 is hsa-miR-3656.miR-4746-3p is hsa-miR-4746-3p, miR-3184-5p is hsa-miR-3184-5p, miR-3937 is hsa-miR-3937, miR-6515-3p is hsa-miR-6515-3p, miR-6132 is hsa-miR-6132, miR-187-5p is hsa-miR-187-5p, miR-7111-5p is hsa-miR-7111-5p, miR-5787 is hsa-miR-5787, and miR-6779-5p is hsa-miR-6779-5p, miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4 649-5p, miR-760 is hsa-miR-760, miR-3162-5p is hsa-miR-3162-5p, miR-3178 is hsa-miR-3178, miR-940 is hsa-miR-940, miR-4271 is hsa-miR-4271, miR-6769b-5p is hsa-miR-6769b-5p, miR-4508 is hsa-miR-4508, miR-6826-5p is hsa-miR-6826-5p, miR-6757-5p is hsa-miR-6757-5p, miR-3131 is hsa-miR-3131, and miR-1343-3p is hsa-miR-1343-3p. ,

[0557] Furthermore, in a preferred embodiment of the method of the present invention, specifically, the nucleic acid (specifically, the probe or primer) is selected from the polynucleotides shown in (a) to (e) below.

[0558] (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0559] (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 1–125 and 466–478.

[0560] (c) A polynucleotide, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases, consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 1 to 125, 466 to 478, or a base sequence in which u is t.

[0561] (d) A polynucleotide containing a base sequence complementary to the base sequence shown in any of the sequence numbers 1–125, 466–478, or a base sequence in which u is t, and

[0562] (e) A polynucleotide that hybridizes with any of the polynucleotides (a) to (d) above under strict conditions.

[0563] In the method of the present invention, it is further possible to use miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR- Nucleic acids specifically bound to at least one of the following polynucleotides: miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, miR-6780b-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p.

[0564] In a preferred embodiment, such nucleic acids specifically include: miR-6808-5p as hsa-miR-6808-5p, miR-6774-5p as hsa-miR-6774-5p, miR-4656 as hsa-miR-4656, miR-6806-5p as hsa-miR-6806-5p, miR-1233-5p as hsa-miR-1233-5p, miR-328-5p as hsa-miR-328-5p, miR-4674 as hsa-miR-4674, miR-2110 as hsa-miR-2110, and miR-60... 76 is hsa-miR-6076, miR-3619-3p is hsa-miR-3619-3p, miR-92a-2-5p is hsa-miR-92a-2-5p, miR-128-1-5p is hsa-miR-128-1-5p, miR-638 is hsa-miR-638, miR-2861 is hsa-miR-2861, miR-371a-5p is hsa-miR-371a-5p, miR-211-3p is hsa-miR-211-3p, miR-1273g-3p is hsa-miR-1273g-3p, mi R-1203 is hsa-miR-1203, miR-122-5p is hsa-miR-122-5p, miR-4258 is hsa-miR-4258, miR-4484 is hsa-miR-4484, miR-4648 is hsa-miR-4648, miR-6780b-5p is hsa-miR-6780b-5p, miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4649-5p, miR-760 is hsa-miR-760, and miR-3162-5p is hsa-miR-1203. R-3162-5p, miR-3178 is hsa-miR-3178, miR-940 is hsa-miR-940, miR-4271 is hsa-miR-4271, miR-6769b-5p is hsa-miR-6769b-5p, miR-4508 is hsa-miR-4508, miR-6826-5p is hsa-miR-6826-5p, miR-6757-5p is hsa-miR-6757-5p, miR-3131 is hsa-miR-3131, and miR-1343-3p is hsa-miR-1343-3p.

[0565] Furthermore, in a preferred embodiment, specifically, such nucleic acid is selected from the polynucleotides shown in (f) to (j) below.

[0566] (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0567] (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126–148.

[0568] (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequences 126 to 148 or a base sequence in which u is t, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide containing 15 or more consecutive bases.

[0569] (i) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequences 126–148, or a base sequence in which u is t, and

[0570] (j) A polynucleotide that hybridizes with any of the polynucleotides (f) to (i) above under strict conditions.

[0571] Examples of specimens used in the method of this invention include specimens prepared from the subject's live tissue (preferably bile duct tissue), blood, serum, plasma, urine, and other bodily fluids. Specifically, specimens containing RNA prepared from such tissue; specimens containing polynucleotides prepared from such tissue; and bodily fluids such as blood, serum, plasma, and urine, as well as live tissue collected from the subject through biopsy or surgical removal, can be used to prepare specimens for measurement.

[0572] In this specification, the test subject refers to mammals, such as humans, monkeys, mice, rats, etc. (not limited to humans), with humans being the preferred candidate.

[0573] The steps of the method of the present invention can be modified according to the type of specimen used as the test subject.

[0574] When RNA is used as the assay target, the detection of biliary tract cancer (cells) may include, for example, the following steps (a), (b) and (c):

[0575] (a) The step of binding RNA modulated by the test subject or complementary polynucleotide (cDNA) transcribed therefrom to the polynucleotide in the kit or device of the present invention.

[0576] (b) The step of determining the RNA derived from the sample or the cDNA synthesized from the RNA that binds to the polynucleotide by hybridization using the above-mentioned polynucleotide as a nucleic acid probe or by quantitative RT-PCR using the above-mentioned polynucleotide as a primer.

[0577] (c) Based on the results of the measurements in (b) above, the steps for evaluating the presence or absence of biliary tract cancer (expression of the gene of origin).

[0578] To detect, examine, evaluate, or diagnose biliary tract cancer (expression of genes of origin) in vitro using this invention, various hybridization methods can be used, such as RNA blotting, DNA blotting, RT-PCR, DNA microarray analysis, in situ hybridization, Northern blotting, and Southern blotting.

[0579] In the case of RNA blotting, by using the nucleic acid probes described above that are applicable to this invention, the presence and expression levels of various genes in RNA can be detected and determined. Specifically, the following method can be exemplified: the nucleic acid probe (complementary strand) is subjected to radioactive isotope ( 32 P, 33 P, 35 This method involves labeling the DNA / RNA double strands with substances such as radioactive isotopes or fluorescent substances, hybridizing them with RNA derived from the subject's living tissue that has been transferred to a nylon membrane or similar material using conventional methods, and then detecting and measuring the signal originating from the label (radioactive isotope or fluorescent substance) using a radiation detector (e.g., BAS-1800II (Fujifilm Corporation)) or a fluorescence detector (e.g., STORM 865 (GE HealthCare)).

[0580] In the case of quantitative RT-PCR, the presence and expression level of genes in RNA can be detected and determined by using the primers available in this invention. Specifically, the following method can be exemplified: cDNA is prepared from RNA derived from the living tissue of the test subject using conventional methods, and the cDNA is used as a template to amplify the target gene regions. One pair of primers (composed of a positive and a negative strand that binds to the cDNA) of this invention is hybridized with the cDNA, and PCR is performed using conventional methods to detect the resulting double-stranded DNA. Alternatively, methods for detecting double-stranded DNA may include: performing the above-mentioned PCR using primers pre-labeled with radioactive isotopes or fluorescent substances; electrophoresis of the PCR products on an agarose gel, staining the double-stranded DNA with ethidium bromide, etc., and detecting it; or transferring the generated double-stranded DNA onto a nylon membrane, etc., using conventional methods, and hybridizing it with a labeled nucleic acid probe for detection.

[0581] In the case of nucleic acid array analysis, an RNA chip or DNA chip is used in which the nucleic acid probes (single-stranded or double-stranded) of the present invention are adhered to a substrate (solid phase). The area where the nucleic acid probe is adhered is called a probe spot, and the area where the nucleic acid probe is not adhered is called a blank spot. Substances obtained by immobilizing gene clusters on a substrate are generally referred to as nucleic acid chips, nucleic acid arrays, microarrays, etc. DNA or RNA arrays include DNA or RNA macroarrays and DNA or RNA microarrays, but in the case of chips mentioned in this specification, all of these arrays are included. As a DNA chip, the 3D-Gene (registered trademark) Human miRNA Oligo chip (Toray Industries, Inc.) can be used, but it is not limited to this.

[0582] The determination of DNA chips is not limited. For example, a method can be used to detect and determine the signal from the marker of the nucleic acid probe using an image detector (such as Typhoon 9410 (GE HealthCare), 3D-Gene (registered trademark) scanner (Toray Industries, Inc.)).

[0583] The “strict conditions” used in this specification refer to conditions under which the nucleic acid probe hybridizes to the target sequence to a greater extent than that for other sequences (e.g., a measurement greater than the average of the background measurements plus the standard error of the background measurements × 2).

[0584] The stringent conditions are defined based on the hybridization and subsequent washing conditions. The hybridization conditions are not limited, and can be, for example, 1 to 24 hours at 30°C to 60°C in a solution containing SSC, a surfactant, formamide, dextran sulfate, a blocking agent, etc. Here, 1×SSC is an aqueous solution (pH 7.0) containing 150 mM sodium chloride and 15 mM sodium citrate, and the surfactant includes SDS (sodium dodecyl sulfate), Triton, or Tween, etc. More preferably, the hybridization conditions contain 3 to 10×SSC and 0.1 to 1% SDS. As another condition specifying the stringent conditions, the washing conditions after hybridization can be exemplified by, for example, continuous washing with a solution containing 0.5×SSC and 0.1% SDS at 30°C, a solution containing 0.2×SSC and 0.1% SDS at 30°C, and a solution containing 0.05×SSC at 30°C. The complementary chain is preferably a complementary chain that maintains a hybridization state with the target positive chain even during washing under such conditions. Specifically, examples of such complementary chains include chains composed of base sequences that are in a completely complementary relationship with the base sequence of the target positive chain, and chains composed of base sequences that have at least 80%, preferably at least 85%, more preferably at least 90% or at least 95%, for example at least 98% or at least 99% identity with the chain.

[0585] Other examples of “strict conditions” in these hybridizations, such as those described in Sambrook, J. & Russell, D., Molecular Cloning, A LABORATORY MANUAL, Cold Spring Harbor Laboratory Press, Vol. 1, pp. 7.42–7.45, Vol. 2, pp. 8.9–8.17, published January 15, 2001, may be utilized in this invention.

[0586] Examples of conditions for performing PCR using the polynucleotide fragments in the kit of the present invention as primers include, for example, using a PCR buffer composed of 10 mM Tris-HCl (pH 8.3), 50 mM KCl, and 1–2 mM MgCl2, and processing at a temperature of 5–10°C + the Tm value calculated from the primer sequence for approximately 15 seconds to 1 minute. A method for calculating this Tm value is given as Tm value = 2 × (number of adenine residues + number of thymine residues) + 4 × (number of guanine residues + number of cytosine residues).

[0587] In addition, when using the quantitative RT-PCR method, commercially available assay kits specifically designed for the quantitative determination of miRNAs can be used, such as TaqMan MicroRNAAssays (Life Technologies), LNA-based MicroRNA PCR (Exiqon), and Ncode miRNA qRT-PCT kit (Invitrogen).

[0588] The calculation of gene expression levels is not limited. In this invention, statistical processing described in, for example, *Statistical analysis of gene expression microarray data* (by Speed ​​T., Chapman and Hall / CRC) and *Abeginner's guide to microarray gene expression data analysis* (by Austin HC et al., Blackwell Publishing) can be used. For example, the average value of the blank point measurements on the DNA chip can be added to two times, preferably three times, and more preferably six times the standard deviation of the blank point measurements, and probe points with signal values ​​above this value can be considered as detection points. Furthermore, the average value of the blank point measurements can be considered as background and subtracted from the probe point measurements to obtain the gene expression level. For missing gene expression values, they can be removed from the analysis target, or preferably replaced with the minimum gene expression value in each DNA chip, or more preferably replaced with the value obtained by subtracting 0.1 from the logarithm of the minimum gene expression value. Furthermore, in order to remove genes with low signal intensity, only genes with expression levels of 2 to the power of 6, more preferably 2 to the power of 8, and more preferably 2 to the power of 10 or higher, representing 20% ​​or more of the sample number to be analyzed, can be selected as the analysis targets. The normalization of gene expression levels is not limited, and examples include global normalization and quantile normalization (Bolstad, BM et al., 2003, Bioinformatics, Vol. 19, pp. 185-193).

[0589] The present invention further provides the following method: using the detection polynucleotide, kit, device (e.g., chip), or combination thereof of the present invention, to determine the expression level of a target gene or gene in a specimen derived from a test subject, using the gene expression levels of specimens derived from biliary tract cancer patients and specimens derived from healthy individuals as teacher samples to make a discriminant (discriminant function) to determine or evaluate whether the specimen contains and / or does not contain genes derived from biliary tract cancer.

[0590] That is, the present invention further provides a method comprising the following steps: a first step of in vitro determination of the expression level of a target gene (target nucleic acid) in a plurality of specimens known to contain or not contain a gene of biliary tract cancer origin using the detection polynucleotide, kit, device (e.g., chip), or combination thereof of the present invention; a second step of making a discriminant formula for a teacher sample using the measured value of the expression level of the target gene obtained in the first step; a third step of in vitro determination of the expression level of the target gene in a specimen of test subject origin, similar to the first step; and a fourth step of determining or evaluating whether the specimen contains a gene of biliary tract cancer origin or not, based on the result obtained from the discriminant formula by substituting the measured value of the expression level of the target gene obtained in the third step into the discriminant formula obtained in the second step, wherein the target gene is a gene that can be detected by the detection polynucleotide contained in the polynucleotide, kit, or device (e.g., chip). Here, Fisher's discriminant analysis, nonlinear discriminant analysis using Mahalanobis distance, neural networks, support vector machines (SVMs), etc., can be used to generate discriminants, but are not limited to these.

[0591] Linear discriminant analysis uses Equation 1 as the discriminant to determine the group affiliation when the group boundary is a straight line or a hyperplane. Here, x is the explanatory variable, w is the coefficient of the explanatory variable, and w0 is the constant term.

[0592] [Number 1]

[0593]

[0594] The value obtained from the discriminant can be called the discriminant score. The measured values ​​of the newly given dataset are substituted into the discriminant as explanatory variables, and the sign of the discriminant score is used to distinguish the group.

[0595] Fisher's discriminant analysis, a type of linear discriminant analysis, is a dimensionality reduction method used to select a dimension suitable for class discrimination. It focuses on the variance of composite variables, minimizing the variance of data with the same label to create a composite variable with high discriminative power (Venables, WN et al., Modern Applied Statistics with S. Fourth edition, Springer, 2002). Fisher's discriminant analysis finds the projection direction w that maximizes Equation 2. Here, μ is the mean of the input, ng is the number of data belonging to class g, and μg is the mean of the input for data belonging to class g. The numerator and denominator are the between-group variance and within-group variance, respectively, when the data are projected onto the vector w. The discriminant coefficient wi is obtained by maximizing this ratio (Kanemori et al., "Pattern Understanding," Kyoritsu Publishing (2009); Richard O. et al., Pattern Classification Second Edition, Wiley-Interscience, 2000).

[0596] [Number 2]

[0597]

[0598] Among them, satisfying

[0599] The Mahalanobis distance, calculated using Equation 3 which considers data correlation, can be used in nonlinear discriminant analysis to classify groups based on their Mahalanobis distance. Here, μ is the center vector of each group, and S... -1 This is the inverse of the variance-covariance matrix of the group. The center vector is calculated from the explanatory variable x, and can be the mean vector, center value vector, etc.

[0600] [Number 3]

[0601]

[0602] SVM, or Discriminant Vector Machine, is a discriminant analysis method examined by V. Vapnik (The Nature of Statistical Leaning Theory, Springer, 1995). It uses specific data items from a known dataset as explanatory variables and the groups to be classified as target variables. A boundary surface, called a hyperplane, is determined to correctly classify the dataset into the known groups, and a discriminant is established using this boundary surface. This discriminant can then be used to classify groups by substituting measurements from a newly given dataset as explanatory variables. Furthermore, the result can be the group to be classified, the probability of being classified into that group, or the distance from the hyperplane. For SVM, as a method to handle nonlinear problems, it is known to transform feature vectors into a higher-dimensional nonlinear space and perform linear discrimination in that space. An expression that represents the inner product of two elements in a nonlinearly mapped space using only the inputs from their original spaces is called a kernel. Examples of kernels include linear kernels, RBF (Radial Basis Function) kernels, and Gaussian kernels. By mapping to higher dimensions through kernels, and actually avoiding the computation of features of the mapped space, the optimal discriminant is constructed solely through the computation of kernels, i.e., discriminant (e.g., Hideki Aso et al., Statistical Science Frontier 6 "Pattern Recognition and Learning of New Concepts and Techniques in Statistics", Iwanami Shoten (2004), Nello Cristianini et al., Introduction to SVM, Kyoritsu Publishing (2008)).

[0603] C-Support Vector Classification (C-SVC), a type of SVM method, learns two sets of explanatory variables to create a hyperplane that determines which group an unknown dataset belongs to (C. Cortes et al., 1995, Machine Learning, Vol. 20, pp. 273-297).

[0604] The following shows an example of calculating the C-SVC discriminant that can be used in the method of the present invention. First, all subjects are divided into two groups: patients with biliary tract cancer and healthy individuals. To determine whether a subject is a patient with biliary tract cancer or a healthy individual, a biliary tract tissue examination can be used, for example.

[0605] Next, a dataset consisting of the whole gene expression levels of two separate groups of serum-derived specimens (hereinafter, the learning specimen group) is prepared. A C-SVC-based discriminant is determined, with genes showing a clear difference in gene expression levels between the two groups designated as explanatory variables and the grouping itself designated as the target variable (e.g., -1 and +1). Equation 4 is the objective function for optimization, where e represents the entire input vector, y represents the target variable, a represents the Lagrange indeterminate multiplier vector, Q represents the positive definite matrix, and C represents the parameters for adjusting the constraints.

[0606] [Number 4]

[0607]

[0608] Among them, what satisfies y T a = 0, 0 ≤ a i ≤C,i=1,...,l,

[0609] Equation 5 is the final discriminant, and the group to which a value belongs can be determined by the sign of the value obtained through the discriminant. Here, x is the support vector, y is the label of the displayed group, a is the corresponding coefficient, b is the constant term, and K is the kernel function.

[0610] [Number 5]

[0611]

[0612] As a kernel function, an RBF kernel, as defined in Equation 6, can be used. Here, x represents the support vectors, and γ represents the kernel parameter that adjusts the complexity of the hyperplane.

[0613] [Number 6]

[0614] K(x i ,x j )=exp(-r||x i -x j || 2 ), r < 0 Equation 6

[0615] In addition to these, methods such as neural networks, k-nearest neighbors, decision trees, and logistic regression analysis can be used to determine or evaluate whether a specimen from a subject source contains and / or does not contain the expression of target genes from biliary tract cancer, or to compare and evaluate their expression levels with those from a healthy control.

[0616] The method of the present invention may include, for example, the following steps (a), (b) and (c):

[0617] (a) The step of determining the expression level of the target gene in known specimens, including tissues containing biliary tract cancer-derived genes from biliary tract cancer patients and / or tissues not containing biliary tract cancer-derived genes from healthy individuals, using the detection polynucleotide, kit, or device (e.g., DNA chip) of the present invention.

[0618] (b) The step of constructing the discriminant equations 1-3, 5 and 6 above from the measured values ​​of the expression level obtained by (a).

[0619] (c) The steps of determining the expression level of the target gene in a specimen derived from a test subject using the detection polynucleotide, kit, or device (e.g., DNA chip) of the present invention, substituting the measured value into the discriminant made by (b), and determining or evaluating whether the specimen contains and / or does not contain the target gene derived from biliary tract cancer, or comparing and evaluating the expression level with a control derived from a healthy body.

[0620] Here, x in Equations 1 to 3, 5 and 6 is an explanatory variable, which includes values ​​obtained by measuring polynucleotides or fragments thereof selected from the polynucleotide classes described in Sections 2 above. Specifically, the explanatory variables used to distinguish between biliary tract cancer patients and healthy individuals of the present invention are, for example, gene expression levels selected from (1) to (2) below.

[0621] (1) Gene expression levels in the serum of a patient with biliary tract cancer or a healthy individual, determined by any one of the DNA sequences containing 15 or more consecutive bases of the base sequence shown in any one of the sequence numbers 1 to 125, 466 to 478 or their complementary sequence.

[0622] (2) Gene expression levels in the serum of patients with biliary tract cancer or healthy individuals as determined by any one of the DNA sequences containing 15 or more consecutive bases of the base sequence shown in any one of the sequence numbers 126 to 148 or its complementary sequence.

[0623] As shown above, as a method for determining or evaluating whether a specimen from a test subject contains and / or does not contain genes from biliary tract cancer, the discriminant formula needs to be generated from a study specimen group. In order to improve the accuracy of the discriminant formula, genes that show clear differences between the two groups in the study specimen group need to be used in the discriminant formula.

[0624] Furthermore, the determination of the genes used as explanatory variables in the discriminant is preferably performed as follows. First, the whole gene expression levels of the biliary tract cancer patient group (used as the learning specimen group) and the whole gene expression levels of the healthy group are used as datasets. The p-values ​​of the t-test (as a parametric analysis), the p-values ​​of the Mann-Whitney U test (as a non-parametric analysis), or the p-values ​​of the Wilcoxon test are used to calculate the difference in the expression levels of each gene between the two groups.

[0625] A p-value obtained through the test is considered statistically significant if the risk (significance level) is, for example, less than 5%, 1%, or 0.01%.

[0626] To correct for the increased probability of Type I errors caused by repeated testing, known methods such as Bonferroni and Holm's methods can be used (e.g., Yasushi Nagata et al., "Basic of Statistical Multiple Comparisons", Scientist (2007)). For example, Bonferroni correction involves multiplying the p-value obtained from the test by the number of repetitions, i.e., the number of genes used in the analysis, and comparing it with the desired significance level, thereby suppressing the probability of Type I errors in the overall test.

[0627] Furthermore, instead of testing, the absolute value of the fold change (the median expression ratio) between the gene expression levels in the biliary tract cancer patient group and the healthy group is calculated. This allows selection of the genes used as explanatory variables in the discriminant. Additionally, ROC curves can be plotted using the gene expression levels from both the biliary tract cancer patient group and the healthy group, and the AUROC value can be used as a benchmark for selecting the genes used as explanatory variables in the discriminant.

[0628] Next, using any number of genes with large differences in gene expression levels obtained here, a discriminant can be calculated using the various methods described above. Methods for constructing a discriminant that yields the highest accuracy include: constructing a discriminant using all combinations of genes that satisfy the significance level of the P-value; and repeatedly evaluating the discriminant by adding genes one by one in order of large differences in gene expression levels (Furey TS. et al., 2000, Bioinformatics., Vol. 16, pp. 906-14). For this discriminant, the gene expression levels of other independent biliary tract cancer patients or healthy individuals are substituted into the explanatory variables, and the discrimination result for that independent biliary tract cancer patient or healthy individual is calculated. That is, by evaluating the found diagnostic gene set and the discriminant constructed using the diagnostic gene set against independent sample groups, a more general diagnostic gene set for detecting biliary tract cancer and a method for discriminating biliary tract cancer can be found.

[0629] Furthermore, the split-sample method is preferred for evaluating the discriminant performance (generalization) of this discriminant formula. That is, the dataset is divided into a training sample group and a test sample group. In the training sample group, gene selection and discriminant formulation are performed through statistical tests. The results of the test sample group and the true group to which the test sample group belongs are then used to calculate accuracy, sensitivity, and specificity to evaluate the discriminant performance. Alternatively, the dataset can be used without splitting. Gene selection and discriminant formulation are performed using all samples through statistical tests. Newly prepared samples are then analyzed using the discriminant formula to calculate accuracy, sensitivity, and specificity to evaluate the discriminant performance.

[0630] This invention provides a polynucleotide for detection or disease diagnosis useful in the diagnosis and treatment of biliary tract cancer, a method for detecting biliary tract cancer using the polynucleotide, and a detection kit and apparatus for biliary tract cancer containing the polynucleotide. In particular, in order to implement the selection of diagnostic genes and the creation of discriminant formulas that demonstrate accuracy exceeding that of existing biliary tract cancer diagnostic methods using tumor markers CEA and CA19-9, in the method of this invention, for example, by comparing expressed genes in the serum of patients who, although negative for CEA and CA19-9, ultimately had biliary tract cancer confirmed by sophisticated examinations such as computed tomography using contrast agents, with expressed genes in the serum of patients without biliary tract cancer, a set of diagnostic genes and discriminant formulas demonstrating accuracy exceeding that of CEA and CA19-9 can be constructed.

[0631] For example, any combination of one or more of the aforementioned polynucleotides based on the base sequences or their complementary sequences shown in any of the sequences 1-125 and 466-478, as described above, and, depending on the circumstances, one or more of the aforementioned polynucleotides based on the base sequences or their complementary sequences shown in any of the sequences 126-148, is designated as the diagnostic gene set. Furthermore, a discriminant is constructed using the expression levels of this diagnostic gene set in specimens derived from patients with grade I biliary tract cancer and from healthy individuals with grade II biliary tract cancer. The result is that by measuring the expression levels of this diagnostic gene set in unknown specimens, it is possible to distinguish with up to 100% accuracy whether an unknown specimen contains or does not contain genes derived from biliary tract cancer.

[0632] Example

[0633] The invention will be further illustrated by the following embodiments. However, the scope of the invention is not limited to these embodiments.

[0634] [Reference Example 1]

[0635] <Collection of specimens from patients with biliary tract cancer and healthy individuals>

[0636] Serum was collected from 100 healthy individuals who had obtained informed consent and 67 patients with biliary tract cancer whose primary cancer could not be confirmed outside the bile duct (1 case of stage IA, 8 cases of stage IB, 8 cases of stage II, 3 cases of stage IIA, 5 cases of stage IIB, 14 cases of stage III, 2 cases of stage IIIB, 1 case of stage IVa, and 25 cases of stage IVb). Serum was collected from each group using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.) as a study sample group. Similarly, serum was collected from 50 healthy individuals who had obtained informed consent and 33 patients with biliary tract cancer (1 case of stage 0, 2 cases of stage I, 1 case of stage IA, 2 cases of stage IB, 2 cases of stage II, 5 cases of stage IIA, 4 cases of stage IIB, 5 cases of stage III, 1 case of stage IV, 1 case of stage IVa, and 9 cases of stage IVb) using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.) as the test specimen group.

[0637] <Total RNA Extraction>

[0638] Total RNA was obtained from 300 μL of serum obtained from 250 individuals, including 150 healthy individuals and 100 patients with biliary tract cancer, by combining the aforementioned learning and testing sample groups. The RNA extraction reagent was obtained using RNA extraction reagent from liquid sample kits manufactured by 3D-Gene (registered trademark) (Toray Industries, Inc.) according to the company's protocol.

[0639] <Gene Expression Measurement>

[0640] Total RNA was obtained from the serum of 250 individuals (150 healthy individuals and 100 patients with biliary tract cancer) by combining the aforementioned study and test sample groups. The miRNAs were fluorescently labeled using the 3D-Gene miRNA Labeling kit (Toray Industries, Inc.) according to the company's instructions (ver 2.20). As an oligoDNA chip, the miRNAs in the total RNA were hybridized to the probes on the DNA chip using the 3D-Gene Human miRNA Oligo chip (Toray Industries, Inc.) equipped with probes containing sequences complementary to 2,555 miRNAs registered in miRBase version 20, under stringent conditions according to the company's instructions. Hybridization and subsequent washing were performed. The DNA chip was scanned using a 3D-Gene scanner (Toray Industries, Inc.) to obtain images, and the fluorescence intensity was quantified using 3D-Gene Extraction (Toray Industries, Inc.). The numerically quantified fluorescence intensity was converted to a logarithm with a base of 2 and set as the gene expression level. Blank values ​​were subtracted, and missing values ​​were replaced with the logarithm of the minimum gene expression level from each DNA chip, minus 0.1. The results yielded the gene expression levels of all miRNAs in the serum of 100 patients with biliary tract cancer and 150 healthy individuals. The calculation and statistical analysis of the numerically quantified miRNA gene expression levels were performed using R language 3.0.2 (R Development Core Team (2013). R: A language and environment for statistical computing. R Foundation for Statistical Computing, URL http: / / www.R-project.org / ) and MASS package 7.3-30 (Venables, WN & Ripley, BD (2002) Modern Applied Statistics with S. Fourth Edition. Springer, New York. ISBN 0-387-95457-0).

[0641] [Reference Example 2]

[0642] <Collection of Specimens for Other Cancers and Benign Diseases>

[0643] Thirty-five patients with colorectal cancer, 37 patients with gastric cancer, 32 patients with esophageal cancer, 38 patients with liver cancer, and 13 patients with benign pancreatic and biliary tract diseases who had obtained informed consent had their serum collected using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.). These serum samples were then combined with those from 67 patients with biliary tract cancer (1 case in stage 0, 2 cases in stage I, 1 case in stage IA, 4 cases in stage IB, 8 cases in stage II, 4 cases in stage IIA, 6 cases in stage IIB, 14 cases in stage III, 1 case in stage IIIB, 25 cases in stage IV, and 1 case in stage IVa) from Reference Case 1 and 93 healthy individuals to form a study sample group. Similarly, serum samples were collected from 15 patients with colorectal cancer, 13 patients with gastric cancer, 18 patients with esophageal cancer, 12 patients with liver cancer, and 8 patients with benign pancreatic and biliary tract diseases who had obtained informed consent. Serum samples were collected using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Corporation). These samples were then combined with those from 33 biliary tract cancer patients (1 in stage IA, 6 in stage IB, 2 in stage II, 4 in stage IIA, 3 in stage IIB, 5 in stage III, 1 in stage IIIB, and 11 in stage IV) and 57 healthy individuals from Reference Example 1 to form the test sample group. Total RNA extraction and gene expression level determination and analysis were performed in the same manner as in Reference Example 1.

[0644] [Example 1]

[0645] <Methods for selecting genetic markers from specimens in the learning specimen group and evaluating the discriminative performance of individual genetic markers from specimens in the test specimen group for biliary tract cancer>

[0646] This embodiment investigates a method for evaluating the discriminative performance of selected gene markers for distinguishing between biliary tract cancer and healthy individuals in a study group of specimens, and in separate test specimen groups independent of the study group.

[0647] Specifically, firstly, the miRNA expression levels of the learning and test specimen groups obtained from Reference Example 1 above were merged and normalized using quantile normalization.

[0648] Next, diagnostic genes were selected using the study specimen group. Here, to obtain more reliable diagnostic biomarkers, only genes with gene expression levels of 2 to the power of 6 or higher were selected from either the biliary tract cancer patient group or the healthy body group within the study specimen group. Furthermore, for genes with statistical significance used to differentiate between the biliary tract cancer patient group and the healthy body group, their respective gene expression levels were Bonferroni corrected using a two-tailed t-test assuming equal variance. Genes satisfying p < 0.01 were used as explanatory variables for the discriminant, and are listed in Table 2.

[0649] By operating in this way, the hsa-miR-125a-3p, hsa-miR-6893-5p, hsa-miR-204-3p, hsa-miR-4476, hsa-miR-4294, hsa-miR-150-3p, hsa-miR-6729-5p, hsa-miR-7641, hsa-miR-6765-3p, hsa-miR-6820-5p, hsa-miR-575, hsa-miR-6836-3p, hsa-miR-1469, hsa-miR-663a, hsa-miR-6075, hsa-miR-4634, hsa-miR-423-5p, hsa-miR-4454, hsa-miR-7109-5p, hsa-miR-6789-5p, hsa-miR-6877-5p, hsa-miR-4792, hsa-miR-4530, hsa-miR-7975, hsa-miR-6724-5p, hsa-miR-8073, hsa-miR-7977, hsa-miR-1231, hsa-miR-6799-5p, hsa-miR-615-5p, hsa-miR-4450, hsa-miR-6726-5p, hsa-miR-6875-5p, hsa-miR-4734, hsa-miR-16-5p, hsa-miR-602, hsa-miR-4651, hsa-miR-8069, hsa-miR-1238-5p, hsa-miR-6880-5p, hsa-miR-8072, hsa-miR-4723-5p, hsa-miR-4732-5p, hsa-miR-6125, hsa-miR-6090, hsa-miR-7114-5p, hsa-miR-564, hsa-miR-451a, hsa-miR-3135b, hsa-miR-4497, hsa-miR-4665-5p, hsa-miR-3622a-5p, hsa-miR-6850-5p, hsa-miR-6821-5p, hsa-miR-5100, hsa-miR-6872-3p, hsa-miR-4433-3p, hsa-miR-1227-5p, hsa-miR-3188, hsa-miR-7704, hsa-miR-3185, hsa-miR-1908-3p, hsa-miR-6781-5p, hsa-miR-6805-5p, hsa-miR-8089, hsa-miR-665, hsa-miR-4486, hsa-miR-6722-3p shown by serial numbers 1 to 125 were discovered.Genes such as hsa-miR-1260a, hsa-miR-4707-5p, hsa-miR-6741-5p, hsa-miR-1260b, hsa-miR-1246, hsa-miR-6845-5p, hsa-miR-4638-5p, hsa-miR-6085, hsa-miR-1228-3p, hsa-miR-4534, hsa-miR-5585-3p, hsa-miR-4741, hsa-miR-4433b-3p, hsa-miR-197-5p, hsa-miR-718, hsa-miR-4513, hsa-miR-4446-3p, hsa-miR-619-5p, hsa-miR-6816-5p, hsa-miR-6778-5p, hsa-miR-24-3p, hsa-miR-1915-3p, hsa-miR-4665-3p, hsa-miR-4449, hsa-miR-6889-5p, hsa-miR-486-3p, hsa-miR-7113-3p, hsa-miR-642a-3p, hsa-miR-7847-3p, hsa-miR-6768-5p, hsa-miR-1290, hsa-miR-7108-5p, hsa-miR-92b-5p, hsa-miR-663b, hsa-miR-3940-5p, hsa-miR-4467, hsa-miR-6858-5p, hsa-miR-4417, hsa-miR-3665, hsa-miR-4736, hsa-miR-4687-3p, hsa-miR-1908-5p, hsa-miR-5195-3p, hsa-miR-4286, hsa-miR-3679-3p, hsa-miR-6791-5p, hsa-miR-1202, hsa-miR-3656, hsa-miR-4746-3p, hsa-miR-3184-5p, hsa-miR-3937, hsa-miR-6515-3p, hsa-miR-6132, hsa-miR-187-5p, hsa-miR-7111-5p, hsa-miR-5787 and hsa-miR-6779-5p serve as cholangiocarcinoma markers relative to healthy subjects.

[0650] Furthermore, using the expression levels of these genes as indicators, a discriminant analysis by Fisher was conducted to determine the presence or absence of bile duct cancer. Specifically, among the 125 genes selected from the study sample group, newly discovered polynucleotides consisting of any of the base sequences shown in sequence numbers 1 to 125 were input into Equation 2 to generate the discriminant. The calculated accuracy, sensitivity, and specificity are shown in Table 3. Additionally, the discriminant coefficients and constants at this point are shown in Table 4.

[0651] Using the discriminant formula derived above, the accuracy, sensitivity, and specificity in the test specimen group were calculated, and the discriminant performance of the selected polynucleotide was validated using independent specimens (Table 3). For example, when the expression levels of the base sequence shown in sequence number 1 were compared between healthy individuals (100 people) and biliary tract cancer patients (67 people) in the learning specimen group, the gene expression levels in the biliary tract cancer patient group were significantly lower than those in the healthy group (see reference). Figure 2 (Left), and further, this result was also reproduced in the test specimen group of healthy individuals (50 people) and patients with biliary tract cancer (33 people) (see reference). Figure 2 (Right). Similarly, for the other polynucleotides shown in sequences 2–125, significantly lower (-) or higher (+) gene expression levels were obtained in the biliary tract cancer patient group compared to the healthy group (Table 2), and these results were validated in the test specimen group. Furthermore, for example, regarding the base sequence shown in sequence 1, the hit rate for biliary tract cancer detection was calculated using the threshold (5.69) set in the learning specimen group to distinguish between the two groups. The results showed 33 true positives, 49 true negatives, 1 false positive, and 0 false negatives. Using these values ​​as detection performance, an accuracy of 99%, a sensitivity of 100%, and a specificity of 98% were obtained. The detection performance of all polynucleotides shown in sequences 1–125 was thus calculated and is recorded in Table 3.

[0652] Among the polynucleotides composed of the base sequences shown in Table 2 (sequence numbers 1 to 125), for example, those composed of sequence numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 34, 35, 36, 39, 40, 41, 42, 44, 45, 46, 47... The 62 polynucleotides composed of the base sequences shown in 49, 50, 51, 52, 53, 54, 60, 62, 64, 65, 67, 68, 70, 74, 75, 76, 83, 84, 105, and 107 showed sensitivities of 100%, 97%, 97%, 100%, 84.8%, 90.9%, 87.9%, 90.9%, 66.7%, 87.9%, 93.9%, and 75% respectively in the test sample group. 0.8%, 72.7%, 72.7%, 75.8%, 63.6%, 78.8%, 75.8%, 69.7%, 72.7%, 72.7%, 69.7%, 93.9%, 66.7%, 63.6%, 69.7%, 69.7%, 78.8%, 75.8%, 72.7%, 78.8%, 81.8%, 66.7%, 60.6%, 60.6%, 72.7%, 66.7%, 60.6%, 63.6%, 81.8%, 60.6%, 69.7%, 60.6%, 78.8%, 69.7%, 63.6%, 63.6%, 60.6%, 72.7%, 63.6%, 72.7%, 72.7%, 63.6%, 66.7%, 60.6%, 60.6%, 63.6%, 63.6%, 69.7%, 63.6%, 69.7%, 60.6% (Table 3). Here, based on the comparative examples described later, the sensitivity of the existing biomarker CEA in the test specimen group is 33.3%, and the sensitivity of CA19-9 is 59.4% (Table 5). This demonstrates that in the test specimen group, for example, those with serial numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 2... The 62 polynucleotides composed of the base sequences shown in 8, 29, 30, 31, 34, 35, 36, 39, 40, 41, 42, 44, 45, 46, 47, 49, 50, 51, 52, 53, 54, 60, 62, 64, 65, 67, 68, 70, 74, 75, 76, 83, 84, 105, and 107 can individually distinguish biliary tract cancer with a sensitivity higher than that of the existing blood tumor marker CA19-9.

[0653] Furthermore, for example, a nine-nucleotide polynucleotide sequence consisting of the base sequences shown in sequence numbers 1, 2, 3, 4, 10, 11, 12, 23, and 64 can correctly identify all six biliary tract cancer specimens in the test sample group, representing stage 0 and stage 1 (including IA and IB), as biliary tract cancer. Therefore, these polynucleotides can also detect early-stage biliary tract cancer and contribute to its early diagnosis.

[0654] Furthermore, these polynucleotides can accurately classify tumors occupying the bile ducts, including extrahepatic bile ducts, intrahepatic bile ducts, gallbladder, and papillae, as biliary carcinomas in the tested specimen set. In particular, they can also detect carcinomas of the lower bile duct, papillae, and intrahepatic bile ducts, which are considered to have a poor prognosis and are prone to developing asymptomatically.

[0655] [Example 2]

[0656] <Evaluation Method for the Detection Performance of Biliary Tract Cancer Using a Combination of Multiple Gene Markers from Test Specimens>

[0657] This embodiment investigated a method for evaluating the discriminative performance of the gene marker combinations selected in Example 1. Specifically, Fisher's discriminant analysis was performed on 7,750 combinations of expression levels of any two polynucleotides selected in Example 1, consisting of the base sequences shown in sequence numbers 1 to 125, to construct a discriminant formula for determining the presence or absence of bile duct cancer. Next, the accuracy, sensitivity, and specificity in the test specimen group were calculated using the discriminant formula, and the discriminative performance of the selected polynucleotides was validated using independent specimens. The results of implementing bile duct cancer discrimination in the test specimen group using the above-described combinations of 7,750 polynucleotide expression levels were as follows: For example, when comparing the expression levels of polynucleotides consisting of the base sequences shown in sequence numbers 2 and 4 in healthy individuals (50 people) and bile duct cancer patients (33 people) in the test specimen group, a scatter plot showing a significant separation between the expression levels of the healthy group and the bile duct cancer patient group was obtained in the learning specimen group (see reference). Figure 3 (Left), and this result can also be reproduced in the test specimen group (see left). Figure 3(Right). Similarly, even in other combinations of expression levels of the newly discovered polynucleotides composed of the base sequences shown in sequences 1-125, scatter plots showing significant separation between the expression levels of the healthy body group and the biliary tract cancer patient group were obtained, and these results were validated in the test specimen group. Furthermore, for example, with respect to the base sequences shown in sequences 2 and 4, the hit rate for biliary tract cancer detection was calculated using a function (0 = 5.16x + y + 48.11) set in the learning specimen group to distinguish between the two groups, resulting in 33 true positives, 48 ​​true negatives, 2 false positives, and 0 false negatives. These values ​​yielded an accuracy of 98%, a sensitivity of 100%, and a specificity of 96% as the detection performance. In this way, the detection performance of all combinations of expression levels of the newly discovered polynucleotides composed of the base sequences shown in sequences 1-125 was calculated. As an example, 124 combinations of polynucleotides composed of the base sequence shown in Serial No. 1 and polynucleotides composed of base sequences shown in other Serial No. 1s, along with their detection performance, are described in Table 6. For instance, combinations of polynucleotide expression levels composed of the base sequences shown in Serial No. 1 and Serial No. 7, Serial No. 1 and Serial No. 9, Serial No. 1 and Serial No. 25, and Serial No. 1 and Serial No. 66 all showed a sensitivity of 100% in the test sample group. 6,316 combinations of polynucleotide expression levels with a sensitivity higher than that of the existing marker CA19-9 (75.8%, as shown in Table 5) were obtained in the test sample group. In these combinations, all base sequences 1 to 125 listed in Table 2 obtained in Example 1 were used at least once. This demonstrates that in the test sample group, any two combinations of polynucleotide expression levels composed of the base sequences shown in Serial No. 1 to 125 can distinguish biliary tract cancer with a sensitivity higher than that of CA19-9.

[0658] Furthermore, among the 7,750 combinations of expression levels of any two polynucleotides composed of the base sequences shown in SEQ ID NO. 1 to 125, 1,290 combinations correctly identified all six biliary tract cancer specimens (stages 0 and 1, including stages IA and IB) included in the test sample set as biliary tract cancer. In these 1,290 combinations, the polynucleotide composed of the base sequences shown in SEQ ID NO. 1 to 125 was used at least once. That is, these polynucleotides can also detect early-stage biliary tract cancer, contributing to its early diagnosis.

[0659] Thus, even when combining expression levels of 3, 4, 5, 6, 7, 8, 9, 10, or more polynucleotides composed of the base sequences shown in sequence numbers 1-125, markers for detecting biliary tract cancer with excellent sensitivity can be obtained. For example, for the polynucleotides composed of the base sequences shown in sequence numbers 1-125 selected in Example 1, the detection performance was calculated by using combinations of one or more miRNAs, starting one at a time and increasing from the epistatic miRNA, in descending order of statistical significance (P-value). The results showed that the sensitivity in the test sample group was 100% with 1 miRNA, 100% with 2 miRNAs, 100% with 3 miRNAs, 100% with 5 miRNAs, 100% with 10 miRNAs, 100% with 20 miRNAs, 100% with 50 miRNAs, and 100% with 100 miRNAs. These sensitivities are higher than those of existing blood tumor markers, demonstrating that even combinations of multiple miRNAs can create excellent markers for detecting biliary tract cancer. Here, the combination of multiple miRNAs is not limited to combinations in a statistically significant order as described above; any combination of multiple miRNAs can be used for the detection of biliary tract cancer.

[0660] These results suggest that the complete polynucleotide sequence consisting of the base sequences shown in sequence numbers 1 to 125 is an excellent diagnostic marker for biliary tract cancer.

[0661] [Table 2]

[0662]

[0663]

[0664]

[0665]

[0666] [Table 3]

[0667]

[0668]

[0669]

[0670]

[0671] [Table 4]

[0672]

[0673]

[0674]

[0675]

[0676] [Table 5-1]

[0677] Learning Specimen Group

[0678]

[0679] [Table 5-2]

[0680] Test specimen group

[0681] Specimen name Cancer stage CEA (ng / mL) CA19-9 (U / mL) B02 IB 3.1 17.1 B03 IIB 3.9 12.9 B04 IIA 2.3 15.8 B08 0 2.7 19.8 B15 IVb 13 328.4 B16 II 1.1 9.6 B20 IIB 2.3 189.8 B22 I 7.8 49.2 B23 III 0.8 8.2 B24 IV 11.6 B28 III 2.4 64.9 B30 IVb 194.7 4597 B31 IVb 3.4 483.3 B32 IIB 2.7 35.2 B34 III 1.6 123.5 B36 IVb 2.7 3374 B37 III 5.5 145.1 B41 IB 2 27.8 B42 IIA 7 37.8 B46 IA 2.1 38.8 B53 I 2.5 6.4 B60 IIA 2.5 105.5 B65 IIA 1.7 11.9 B66 IIA 4.6 11.1 B68 IIB 1.1 7.2 B70 II 1.6 123.5 B71 IVa 6.5 925 B76 IVb 1482 15.6 B79 IVb 65 6510 B80 IVb 5 229.9 B84 III 3.1 52.5 B88 IVb 76.9 777 P91 IVb 2.3 4308 Sensitivity (%) 33.3 59.4

[0682] In Table 5, CEA values ​​below 5 ng / ml are marked as "-", CA19-9 values ​​below 37 U / ml are marked as "-", and values ​​exceeding these values ​​are marked as "+".

[0683] [Table 6]

[0684]

[0685]

[0686]

[0687]

[0688] [Example 3]

[0689] <Methods for selecting genetic markers when using all samples and evaluating the biliary tract cancer discrimination performance of the obtained genetic markers>

[0690] In this embodiment, the specimens from the learning specimen group and the test specimen group used in Embodiments 1 and 2 above are combined to use all specimens for the selection of gene markers and the evaluation of their biliary tract cancer discrimination performance.

[0691] Specifically, the miRNA expression levels in the serum of 100 patients with biliary tract cancer and 150 healthy individuals obtained from Reference Example 1 were normalized using quantile normalization. To obtain more reliable diagnostic biomarkers, gene biomarkers were selected only those genes with an expression level of 2 to the power of 6 or higher in more than 50% of the samples from either the biliary tract cancer patient group or the healthy individual group. Furthermore, to obtain statistical significance for distinguishing between the biliary tract cancer patient group and the healthy individual group, the p-values ​​obtained from a two-tailed t-test assuming equal variance were Bonferroni corrected, and genes satisfying p < 0.01 were selected as explanatory variables for the discriminant. The gene biomarkers used are listed in Table 7. In addition to the genes listed in Table 2, the following genes were also found: hsa-miR-6808-5p, hsa-miR-6774-5p, hsa-miR-4656, hsa-miR-6806-5p, hsa-miR-1233-5p, hsa-miR-328-5p, hsa-miR-4674, hsa-miR-2110, hsa-miR-6076, hsa-miR-3619-3p, and hsa-miR-92a-2 (as shown in sequence numbers 126–148). The genes hsa-miR-128-1-5p, hsa-miR-638, hsa-miR-2861, hsa-miR-371a-5p, hsa-miR-211-3p, hsa-miR-1273g-3p, hsa-miR-1203, hsa-miR-122-5p, hsa-miR-4258, hsa-miR-4484, hsa-miR-4648, and hsa-miR-6780b-5p were used as biomarkers for biliary tract cancer relative to healthy individuals. Similar to the polynucleotides shown in sequences 1–125, significantly lower (-) or higher (+) expression levels were also observed in the biliary tract cancer patient group compared to the healthy group in the polynucleotides shown in sequences 126–148 (Table 7). These results were validated in the test specimen group. Newly obtained specimens can be identified by using the gene expression level measurements recorded in Table 7 alone or in combination with the gene expression level measurements recorded in Table 2, according to the methods described in Examples 1 and 2.

[0692] [Table 7]

[0693]

[0694]

[0695]

[0696]

[0697] [Example 4]

[0698] <An evaluation method for the discriminative performance of bile duct cancer specificity using a combination of multiple genetic markers from test specimens>

[0699] In this embodiment, the study specimen group described in Reference Example 2 was used as the subject. Using the same method as described in Example 1, the gene expression levels of miRNAs in the serum of biliary tract cancer patients were compared with those of a control group consisting of healthy individuals, colorectal cancer patients, gastric cancer patients, esophageal cancer patients, liver cancer patients, and patients with benign pancreatic and biliary tract diseases. Additional diagnostic gene markers were then selected. The results showed that the specificity of biliary tract cancer was evaluated using one or more markers selected from groups that combined the selected additional diagnostic gene markers (serial numbers 466-478; see Table 1) with the gene markers selected in Example 1.

[0700] Specifically, firstly, the miRNA expression levels of the learning and test specimen groups obtained from Reference Example 2 were merged and normalized using quantile normalization. Next, Fisher's discriminant analysis was performed on combinations of 1 to 4 expression level measurements of any of the polynucleotides consisting of at least one base sequence represented by sequence numbers 1-148 and 466-478 to construct a discriminant for the presence or absence of biliary tract cancer. Next, the biliary tract cancer patient group was used as the positive specimen group, and the healthy body group, colorectal cancer patient group, gastric cancer patient group, esophageal cancer patient group, liver cancer patient group, and pancreatic biliary benign disease patient group were used as the negative specimen groups. The accuracy, sensitivity, and specificity of the test specimen group were calculated using the discriminant constructed above, and the discriminant performance of the selected polynucleotide was validated using independent specimens.

[0701] The polynucleotides composed of the base sequences shown above (sequence numbers 1-148 and 466-478 corresponding to the miRNA markers in Table 1) or their complementary sequences can not only provide high accuracy, sensitivity, and specificity in the determination of the presence or absence of biliary tract cancer, but also specifically distinguish biliary tract cancer from other cancers. Combinations of multiple polynucleotides, including those selected from groups consisting of polynucleotides with base sequences or complementary sequences shown in sequence numbers 1, 4, 5, 11, 12, 15, 23, 29, 39, 40, 54, 76, 79, 91, 103, 115, 121, 134, 143, 466, 469, 472, 473, and 474 (cancer type-specific polynucleotide group 1), and including at least one polynucleotide selected from groups consisting of polynucleotides with base sequences or complementary sequences shown in sequence numbers 4, 5, 12, 15, and 40 (cancer type-specific polynucleotide group 2), can specifically identify biliary tract cancer from other cancers with high accuracy.

[0702] The number of combinations of the aforementioned cancer-type specific polynucleotides can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more, and the discrimination accuracy can be shown to be above 80% in combinations of 4 or more.

[0703] Specifically, the discrimination accuracy when a polynucleotide consisting of the base sequence shown in Serial No. 4 or its complementary sequence is used is as follows. When a single polynucleotide consisting of the base sequence shown in Serial No. 4 or its complementary sequence is used, the accuracy is 81.9% in the learning sample group and 76.9% in the test sample group (Table 8). Furthermore, for example, when a combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 4 or its complementary sequence is used, the accuracy is as high as 86.0% in the learning sample group and 85.3% in the test sample group (Table 9; in the table, "Serial No." indicates a combination of the serial numbers of the two polynucleotides used). Furthermore, for example, when the assay was performed using a combination of three polynucleotides comprising at least one polynucleotide consisting of the base sequence shown in Serial Number 4 or its complementary sequence, the accuracy was highest at 89.5% in the learning sample group and 90.4% in the test sample group (Table 10; "Serial Number" in the table refers to the combination of the sequence numbers of the three polynucleotides used). Furthermore, for example, when the assay was performed using a combination of four polynucleotides comprising at least one polynucleotide consisting of the base sequence shown in Serial Number 4 or its complementary sequence, the accuracy was highest at 91.1% in the learning sample group and 92.3% in the test sample group (Table 11; "Serial Number" in the table refers to the combination of the sequence numbers of the four polynucleotides used).

[0704] Specifically, the discrimination accuracy when a polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence is used is as follows. When a single polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence is used, the accuracy is 79.0% in the learning sample group and 80.8% in the test sample group (Table 8). Furthermore, for example, when a combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence is used, the accuracy is as high as 81.9% in the learning sample group and 86.5% in the test sample group (Table 9). Furthermore, for example, when a combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence is used, the accuracy is as high as 87.6% in the learning sample group and 89.7% in the test sample group (Table 10). Furthermore, for example, when the assay was performed using a combination of four polynucleotides comprising at least one polynucleotide consisting of the base sequence shown in Serial No. 5 or its complementary sequence, the accuracy was up to 93.0% in the learning sample group and 91.0% in the test sample group (Table 11).

[0705] Specifically, the discrimination accuracy when a polynucleotide consisting of the base sequence shown in Serial No. 12 or its complementary sequence is used is as follows. When a single polynucleotide consisting of the base sequence shown in Serial No. 12 or its complementary sequence is used, the accuracy is 80.6% in the learning sample group and 76.9% in the test sample group (Table 8). Furthermore, for example, when a combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 12 or its complementary sequence is used, the accuracy is up to 86.3% in the learning sample group and 85.9% in the test sample group (Table 9). Furthermore, for example, when a combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 12 or its complementary sequence is used, the accuracy is up to 90.2% in the learning sample group and 91.7% in the test sample group (Table 10). Furthermore, for example, when the assay was performed using a combination of four polynucleotides comprising at least one polynucleotide consisting of the base sequence shown in Serial No. 12 or its complementary sequence, the accuracy was up to 93.0% in the learning sample group and 94.2% in the test sample group (Table 11).

[0706] Specifically, the discrimination accuracy when a polynucleotide consisting of the base sequence shown in Serial No. 15 or its complementary sequence is used is as follows. When a single polynucleotide consisting of the base sequence shown in Serial No. 15 or its complementary sequence is used for determination, the accuracy is 83.8% in the learning sample group and 84.0% in the test sample group (Table 8). Furthermore, for example, when a combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 15 or its complementary sequence is used for determination, the accuracy is up to 89.5% in the learning sample group and 89.1% in the test sample group (Table 9). Furthermore, for example, when a combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial No. 15 or its complementary sequence is used for determination, the accuracy is up to 90.5% in the learning sample group and 92.3% in the test sample group (Table 10). Furthermore, for example, when the assay was performed using a combination of four polynucleotides comprising at least one polynucleotide consisting of the base sequence shown in Serial No. 15 or its complementary sequence, the accuracy was up to 93.0% in the learning sample group and 94.2% in the test sample group (Table 11).

[0707] Specifically, the discrimination accuracy when a polynucleotide consisting of the base sequence shown in Serial Number 40 or its complementary sequence is used is as follows. When a single polynucleotide consisting of the base sequence shown in Serial Number 40 or its complementary sequence is used, the accuracy is 80.0% in the learning sample group and 76.9% in the test sample group (Table 8). Furthermore, for example, when a combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial Number 40 or its complementary sequence is used, the accuracy is up to 81.9% in the learning sample group and 86.5% in the test sample group (Table 9). Furthermore, for example, when a combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in Serial Number 40 or its complementary sequence is used,...

Claims

1. Capable of binding to a selected from miR-6893-5p, miR-4476, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4651, miR-8069, miR-1238-5p, miR-6880-5p, miR-8072, miR-4723-5p, miR-4732-5p, miR-6125, miR-6090, miR-7114-5p, miR-564, miR-451a, miR-3135b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3p, miR-1227-5p, miR-3188, miR-7704, miR-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a, miR-4707-5p, miR-6741-5p, miR-1260b, miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-4433b-3p, miR-197-5p, miR-718, miR-4513, miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449, miR-6889-5p, miR-486-3p, miR-7113-3p, miR-642a-3p, miR-7847-3p, which is a biomarker for cholangiocarcinomamiR-6768-5p,miR-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR-4467,miR-6858-5p,miR-4417,miR-3665,miR-4736,miR-4687- 3p,miR-1908-5p,miR-5195-3p,miR-4286,miR-3679-3p,miR-6791-5p,miR-1202,miR-3656,miR-4746-3p,miR-3184-5p,miR-3937,miR-6515-3p, The purpose of using nucleic acid probes that specifically bind to at least one polynucleotide selected from miR-6132, miR-187-5p, miR-7111-5p, miR-5787, miR-6779-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p, and / or primers that specifically recognize and amplify the polynucleotide, for the manufacture of a kit for the detection of biliary tract cancer.

2. The use according to claim 1, wherein, miR-6893-5p is hsa-miR-6893-5p, miR-4476 is hsa-miR-4476, miR-150-3p is hsa-miR-150-3p, miR-6729-5p is hsa-miR-6729-5p, miR-7641 is hsa-miR-7641, miR-6765-3p is hsa-miR-6765-3p, miR-6820-5p is hsa-miR-6820-5p, miR-575 is hsa-miR-575, miR-1469 is hsa-miR-1469, and miR-663a is hsa-miR-663a. miR-6075 is hsa-miR-6075, miR-4634 is hsa-miR-4634, miR-423-5p is hsa-miR-423-5p, miR-4454 is hsa-miR-4454, miR-7109-5p is hsa-miR-7109-5p, miR-6789-5p is hsa-miR-6789-5p, miR-6877-5p is hsa-miR-6877-5p, miR-4792 is hsa-miR-4792, miR-4530 is hsa-miR-4530, miR-7975 is hsa-miR-7975, mi R-6724-5p is hsa-miR-6724-5p, miR-8073 is hsa-miR-8073, miR-7977 is hsa-miR-7977, miR-1231 is hsa-miR-1231, miR-6799-5p is hsa-miR-6799-5p, miR-615-5p is hsa-miR-615-5p, miR-4450 is hsa-miR-4450, miR-6726-5p is hsa-miR-6726-5p, miR-6875-5p is hsa-miR-6875-5p, and miR-4734 is hsa-miR-4734. miR-16-5p is hsa-miR-16-5p, miR-602 is hsa-miR-602, miR-4651 is hsa-miR-4651, miR-8069 is hsa-miR-8069, miR-1238-5p is hsa-miR-1238-5p, miR-6880-5p is hsa-miR-6880-5p, miR-8072 is hsa-miR-8072, miR-4723-5p is hsa-miR-4723-5p, miR-4732-5p is hsa-miR-4732-5p, and miR-6125 is hsa-miR-6125.miR-6090 is hsa-miR-6090, miR-7114-5p is hsa-miR-7114-5p, miR-564 is hsa-miR-564, miR-451a is hsa-miR-451a, miR-3135b is hsa-miR-3135b, miR-4497 is hsa-miR-4497, miR-4665-5p is hsa-miR-4665-5p, miR-3622a-5p is hsa-miR-3622a-5p, miR-6850-5p is hsa-miR-6850-5p, and miR-6821-5p is hsa-miR-6 821-5p, miR-5100 is hsa-miR-5100, miR-6872-3p is hsa-miR-6872-3p, miR-4433-3p is hsa-miR-4433-3p, miR-1227-5p is hsa-miR-1227-5p, miR-3188 is hsa-miR-3188, miR-7704 is hsa-miR-7704, miR-3185 is hsa-miR-3185, miR-1908-3p is hsa-miR-1908-3p, miR-6781-5p is hsa-miR-6781-5p, miR-6805-5 p is hsa-miR-6805-5p, miR-8089 is hsa-miR-8089, miR-665 is hsa-miR-665, miR-4486 is hsa-miR-4486, miR-6722-3p is hsa-miR-6722-3p, miR-1260a is hsa-miR-1260a, miR-4707-5p is hsa-miR-4707-5p, miR-6741-5p is hsa-miR-6741-5p, miR-1260b is hsa-miR-1260b, miR-1246 is hsa-miR-1246, miR-6845- 5p is hsa-miR-6845-5p, miR-4638-5p is hsa-miR-4638-5p, miR-6085 is hsa-miR-6085, miR-1228-3p is hsa-miR-1228-3p, miR-4534 is hsa-miR-4534, miR-5585-3p is hsa-miR-5585-3p, miR-4741 is hsa-miR-4741, miR-4433b-3p is hsa-miR-4433b-3p, miR-197-5p is hsa-miR-197-5p, and miR-718 is hsa-miR-718.miR-4513 is hsa-miR-4513, miR-4446-3p is hsa-miR-4446-3p, miR-619-5p is hsa-miR-619-5p, miR-6816-5p is hsa-miR-6816-5p, miR-6778-5p is hsa-miR-6778-5p, miR-24-3p is hsa-miR-24-3p, miR-1915-3p is hsa-miR-1915-3p, miR-4665-3p is hsa-miR-4665-3p, miR-4449 is hsa-miR-4449, miR-6889-5 p is hsa-miR-6889-5p, miR-486-3p is hsa-miR-486-3p, miR-7113-3p is hsa-miR-7113-3p, miR-642a-3p is hsa-miR-642a-3p, miR-7847-3p is hsa-miR-7847-3p, miR-6768-5p is hsa-miR-6768-5p, miR-1290 is hsa-miR-1290, miR-7108-5p is hsa-miR-7108-5p, miR-92b-5p is hsa-miR-92b-5p, miR-663b is hsa -miR-663b, miR-3940-5p is hsa-miR-3940-5p, miR-4467 is hsa-miR-4467, miR-6858-5p is hsa-miR-6858-5p, miR-4417 is hsa-miR-4417, miR-3665 is hsa-miR-3665, miR-4736 is hsa-miR-4736, miR-4687-3p is hsa-miR-4687-3p, miR-1908-5p is hsa-miR-1908-5p, miR-5195-3p is hsa-miR-5195-3p, miR-42 86 is hsa-miR-4286, miR-3679-3p is hsa-miR-3679-3p, miR-6791-5p is hsa-miR-6791-5p, miR-1202 is hsa-miR-1202, miR-3656 is hsa-miR-3656, miR-4746-3p is hsa-miR-4746-3p, miR-3184-5p is hsa-miR-3184-5p, miR-3937 is hsa-miR-3937, miR-6515-3p is hsa-miR-6515-3p, and miR-6132 is hsa-miR-6132.miR-187-5p is hsa-miR-187-5p, miR-7111-5p is hsa-miR-7111-5p, miR-5787 is hsa-miR-5787, miR-6779-5p is hsa-miR-6779-5p, miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4649-5p, miR-760 is hsa-miR-760, miR-3162-5p is hsa-miR-3162-5p, miR-3178 is h The following are listed: miR-3178 (sa-miR-3178), miR-940 (hsa-miR-940), miR-4271 (hsa-miR-4271), miR-6769b-5p (hsa-miR-6769b-5p), miR-4508 (hsa-miR-4508), miR-6826-5p (hsa-miR-6826-5p), miR-6757-5p (hsa-miR-6757-5p), miR-3131 (hsa-miR-3131), and miR-1343-3p (hsa-miR-1343-3p).

3. According to claim 1, the nucleic acid probe capable of specifically binding to the polynucleotide and / or the primer specifically recognizing and amplifying the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a) to (e) below. (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t. (c) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (d) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t. (e) A polynucleotide that hybridizes with any of the polynucleotides in (a) to (d) under stringent conditions.

4. The use according to any one of claims 1 to 3, wherein the kit further comprises a component capable of reacting with other biliary tract cancer markers selected from miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p, A nucleic acid probe that specifically binds to at least one polynucleotide from miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, and miR-6780b-5p, and / or a primer that specifically recognizes and amplifies that polynucleotide.

5. The use according to claim 4, wherein, miR-6808-5p is hsa-miR-6808-5p, miR-6774-5p is hsa-miR-6774-5p, miR-4656 is hsa-miR-4656, miR-6806-5p is hsa-miR-6806-5p, miR-1233-5p is hsa-miR-1233-5p, miR-328-5p is h sa-miR-328-5p, miR-4674 is hsa-miR-4674, miR-2110 is hsa-miR-2110, miR-6076 is hsa-miR-6076, miR-3619-3p is hsa-miR-3619-3p, miR-92a-2-5p is hsa-miR-92a-2-5p, miR-128- 1-5p is hsa-miR-128-1-5p, miR-638 is hsa-miR-638, miR-2861 is hsa-miR-2861, miR-371a-5p is hsa-miR-371a-5p, miR-211-3p is hsa-miR-211-3p, and miR-1273g-3p is hsa-miR-1273g-3. p, miR-1203 is hsa-miR-1203, miR-122-5p is hsa-miR-122-5p, miR-4258 is hsa-miR-4258, miR-4484 is hsa-miR-4484, miR-4648 is hsa-miR-4648, and miR-6780b-5p is hsa-miR-6780b-5p.

6. According to claim 4, the nucleic acid probe capable of specifically binding to the polynucleotide and / or the primer specifically recognizing and amplifying the polynucleotide is a polynucleotide selected from the polynucleotides shown in (f) to (j) below. (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126 to 148, or a base sequence in which u is t. (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 126 to 148 or the base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (i) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequences 126–148 or a base sequence in which u is t, and (j) A polynucleotide that hybridizes with any of the polynucleotides in (f) to (i) under strict conditions.

7. The use according to claim 1 or 4, wherein the kit comprises at least two nucleic acid probes capable of specifically binding to at least two polynucleotides selected from all biliary tract cancer markers described in claim 1 or 4 and / or at least two primers specifically recognizing and amplifying the polynucleotides.

8. Can bind to a selected from miR-6893-5p, miR-4476, miR-150-3p, miR-6729-5p, miR-7641, miR-6765-3p, miR-6820-5p, miR-575, miR-1469, miR-663a, miR-6075, miR-4634, miR-423-5p, miR-4454, miR-7109-5p, miR-6789-5p, miR-6877-5p, miR-4792, miR-4530, miR-7975, miR-6724-5p, miR-8073, miR-7977, miR-1231, miR-6799-5p, miR-615-5p, miR-4450, miR-6726-5p, miR-6875-5p, miR-4734, miR-16-5p, miR-602, miR-4651, miR-8069, miR-1238-5p, miR-6880-5p, miR-8072, miR-4723-5p, miR-4732-5p, miR-6125, miR-6090, miR-7114-5p, miR-564, miR-451a, miR-3135b, miR-4497, miR-4665-5p, miR-3622a-5p, miR-6850-5p, miR-6821-5p, miR-5100, miR-6872-3p, miR-4433-3p, miR-1227-5p, miR-3188, miR-7704, miR-3185, miR-1908-3p, miR-6781-5p, miR-6805-5p, miR-8089, miR-665, miR-4486, miR-6722-3p, miR-1260a, miR-4707-5p, miR-6741-5p, miR-1260b, miR-1246, miR-6845-5p, miR-4638-5p, miR-6085, miR-1228-3p, miR-4534, miR-5585-3p, miR-4741, miR-4433b-3p, miR-197-5p, miR-718, miR-4513, miR-4446-3p, miR-619-5p, miR-6816-5p, miR-6778-5p, miR-24-3p, miR-1915-3p, miR-4665-3p, miR-4449, miR-6889-5p, miR-486-3p, miR-7113-3p, miR-642a-3p, miR-7847-3p, which are markers for cholangiocarcinomamiR-6768-5p,miR-1290,miR-7108-5p,miR-92b-5p,miR-663b,miR-3940-5p,miR-4467,miR-6858-5p,miR-4417,miR-3665,miR-4736,miR-4687- 3p,miR-1908-5p,miR-5195-3p,miR-4286,miR-3679-3p,miR-6791-5p,miR-1202,miR-3656,miR-4746-3p,miR-3184-5p,miR-3937,miR-6515-3p, The purpose of using nucleic acid probes that specifically bind to at least one polynucleotide selected from miR-6132, miR-187-5p, miR-7111-5p, miR-5787, miR-6779-5p, miR-4516, miR-4649-5p, miR-760, miR-3162-5p, miR-3178, miR-940, miR-4271, miR-6769b-5p, miR-4508, miR-6826-5p, miR-6757-5p, miR-3131, and miR-1343-3p, and / or primers that specifically recognize and amplify the polynucleotide, for the manufacture of a device for the detection of biliary tract cancer.

9. The use according to claim 8, wherein, miR-6893-5p is hsa-miR-6893-5p, miR-4476 is hsa-miR-4476, miR-150-3p is hsa-miR-150-3p, miR-6729-5p is hsa-miR-6729-5p, miR-7641 is hsa-miR-7641, miR-6765-3p is hsa-miR-6765-3p, miR-6820-5p is hsa-miR-6820-5p, miR-575 is hsa-miR-575, miR-1469 is hsa-miR-1469, and miR-663a is hsa-miR-663a. miR-6075 is hsa-miR-6075, miR-4634 is hsa-miR-4634, miR-423-5p is hsa-miR-423-5p, miR-4454 is hsa-miR-4454, miR-7109-5p is hsa-miR-7109-5p, miR-6789-5p is hsa-miR-6789-5p, miR-6877-5p is hsa-miR-6877-5p, miR-4792 is hsa-miR-4792, miR-4530 is hsa-miR-4530, miR-7975 is hsa-miR-7975, mi R-6724-5p is hsa-miR-6724-5p, miR-8073 is hsa-miR-8073, miR-7977 is hsa-miR-7977, miR-1231 is hsa-miR-1231, miR-6799-5p is hsa-miR-6799-5p, miR-615-5p is hsa-miR-615-5p, miR-4450 is hsa-miR-4450, miR-6726-5p is hsa-miR-6726-5p, miR-6875-5p is hsa-miR-6875-5p, and miR-4734 is hsa-miR-4734. miR-16-5p is hsa-miR-16-5p, miR-602 is hsa-miR-602, miR-4651 is hsa-miR-4651, miR-8069 is hsa-miR-8069, miR-1238-5p is hsa-miR-1238-5p, miR-6880-5p is hsa-miR-6880-5p, miR-8072 is hsa-miR-8072, miR-4723-5p is hsa-miR-4723-5p, miR-4732-5p is hsa-miR-4732-5p, and miR-6125 is hsa-miR-6125.miR-6090 is hsa-miR-6090, miR-7114-5p is hsa-miR-7114-5p, miR-564 is hsa-miR-564, miR-451a is hsa-miR-451a, miR-3135b is hsa-miR-3135b, miR-4497 is hsa-miR-4497, miR-4665-5p is hsa-miR-4665-5p, miR-3622a-5p is hsa-miR-3622a-5p, miR-6850-5p is hsa-miR-6850-5p, and miR-6821-5p is hsa-miR-6 821-5p, miR-5100 is hsa-miR-5100, miR-6872-3p is hsa-miR-6872-3p, miR-4433-3p is hsa-miR-4433-3p, miR-1227-5p is hsa-miR-1227-5p, miR-3188 is hsa-miR-3188, miR-7704 is hsa-miR-7704, miR-3185 is hsa-miR-3185, miR-1908-3p is hsa-miR-1908-3p, miR-6781-5p is hsa-miR-6781-5p, miR-6805-5 p is hsa-miR-6805-5p, miR-8089 is hsa-miR-8089, miR-665 is hsa-miR-665, miR-4486 is hsa-miR-4486, miR-6722-3p is hsa-miR-6722-3p, miR-1260a is hsa-miR-1260a, miR-4707-5p is hsa-miR-4707-5p, miR-6741-5p is hsa-miR-6741-5p, miR-1260b is hsa-miR-1260b, miR-1246 is hsa-miR-1246, miR-6845- 5p is hsa-miR-6845-5p, miR-4638-5p is hsa-miR-4638-5p, miR-6085 is hsa-miR-6085, miR-1228-3p is hsa-miR-1228-3p, miR-4534 is hsa-miR-4534, miR-5585-3p is hsa-miR-5585-3p, miR-4741 is hsa-miR-4741, miR-4433b-3p is hsa-miR-4433b-3p, miR-197-5p is hsa-miR-197-5p, and miR-718 is hsa-miR-718.miR-4513 is hsa-miR-4513, miR-4446-3p is hsa-miR-4446-3p, miR-619-5p is hsa-miR-619-5p, miR-6816-5p is hsa-miR-6816-5p, miR-6778-5p is hsa-miR-6778-5p, miR-24-3p is hsa-miR-24-3p, miR-1915-3p is hsa-miR-1915-3p, miR-4665-3p is hsa-miR-4665-3p, miR-4449 is hsa-miR-4449, miR-6889-5 p is hsa-miR-6889-5p, miR-486-3p is hsa-miR-486-3p, miR-7113-3p is hsa-miR-7113-3p, miR-642a-3p is hsa-miR-642a-3p, miR-7847-3p is hsa-miR-7847-3p, miR-6768-5p is hsa-miR-6768-5p, miR-1290 is hsa-miR-1290, miR-7108-5p is hsa-miR-7108-5p, miR-92b-5p is hsa-miR-92b-5p, miR-663b is hsa -miR-663b, miR-3940-5p is hsa-miR-3940-5p, miR-4467 is hsa-miR-4467, miR-6858-5p is hsa-miR-6858-5p, miR-4417 is hsa-miR-4417, miR-3665 is hsa-miR-3665, miR-4736 is hsa-miR-4736, miR-4687-3p is hsa-miR-4687-3p, miR-1908-5p is hsa-miR-1908-5p, miR-5195-3p is hsa-miR-5195-3p, miR-42 86 is hsa-miR-4286, miR-3679-3p is hsa-miR-3679-3p, miR-6791-5p is hsa-miR-6791-5p, miR-1202 is hsa-miR-1202, miR-3656 is hsa-miR-3656, miR-4746-3p is hsa-miR-4746-3p, miR-3184-5p is hsa-miR-3184-5p, miR-3937 is hsa-miR-3937, miR-6515-3p is hsa-miR-6515-3p, and miR-6132 is hsa-miR-6132.miR-187-5p is hsa-miR-187-5p, miR-7111-5p is hsa-miR-7111-5p, miR-5787 is hsa-miR-5787, and miR-6779-5p is hsa-miR-6779-5p. miR-4516 is hsa-miR-4516, miR-4649-5p is hsa-miR-4649-5p, miR-760 is hsa-miR-760, miR-3162-5p is hsa-miR-3162-5p, and miR-3178 is hsa-miR-187-5p. The following are listed: miR-3178 (sa-miR-3178), miR-940 (hsa-miR-940), miR-4271 (hsa-miR-4271), miR-6769b-5p (hsa-miR-6769b-5p), miR-4508 (hsa-miR-4508), miR-6826-5p (hsa-miR-6826-5p), miR-6757-5p (hsa-miR-6757-5p), miR-3131 (hsa-miR-3131), and miR-1343-3p (hsa-miR-1343-3p).

10. The use according to claim 8, wherein the nucleic acid probe capable of specifically binding to the polynucleotide and / or the primer specifically recognizing and amplifying the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a) to (e) below, (a) A polynucleotide consisting of a base sequence represented by any one of sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (b) A polynucleotide containing the base sequence shown in any of the sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t. (c) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (d) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequence numbers 2, 4, 6–11, 13–125, and 466–478, or a base sequence in which u is t. (e) A polynucleotide that hybridizes with any of the polynucleotides in (a) to (d) under stringent conditions.

11. The use according to any one of claims 8 to 10, wherein the device further comprises the ability to interact with other biliary tract cancer markers selected from miR-6808-5p, miR-6774-5p, miR-4656, miR-6806-5p, miR-1233-5p, miR-328-5p, miR-4674, miR-2110, miR-6076, miR-3619-3p, miR-92a-2-5p. A nucleic acid probe that specifically binds to at least one of the polynucleotides selected from miR-128-1-5p, miR-638, miR-2861, miR-371a-5p, miR-211-3p, miR-1273g-3p, miR-1203, miR-122-5p, miR-4258, miR-4484, miR-4648, and miR-6780b-5p, and / or a primer that specifically recognizes and amplifies the polynucleotide.

12. The use according to claim 11, wherein, miR-6808-5p is hsa-miR-6808-5p, miR-6774-5p is hsa-miR-6774-5p, miR-4656 is hsa-miR-4656, miR-6806-5p is hsa-miR-6806-5p, miR-1233-5p is hsa-miR-1233-5p, miR-328-5p is h sa-miR-328-5p, miR-4674 is hsa-miR-4674, miR-2110 is hsa-miR-2110, miR-6076 is hsa-miR-6076, miR-3619-3p is hsa-miR-3619-3p, miR-92a-2-5p is hsa-miR-92a-2-5p, miR-128- 1-5p is hsa-miR-128-1-5p, miR-638 is hsa-miR-638, miR-2861 is hsa-miR-2861, miR-371a-5p is hsa-miR-371a-5p, miR-211-3p is hsa-miR-211-3p, and miR-1273g-3p is hsa-miR-1273g-3. p, miR-1203 is hsa-miR-1203, miR-122-5p is hsa-miR-122-5p, miR-4258 is hsa-miR-4258, miR-4484 is hsa-miR-4484, miR-4648 is hsa-miR-4648, and miR-6780b-5p is hsa-miR-6780b-5p.

13. According to claim 11, the nucleic acid probe capable of specifically binding to the polynucleotide and / or the primer specifically recognizing and amplifying the polynucleotide is a polynucleotide selected from the polynucleotides shown in (f) to (j) below. (f) A polynucleotide consisting of the base sequence shown in any of the sequence numbers 126 to 148, or a sequence of bases in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (g) A polynucleotide containing the base sequence shown in any of the sequence numbers 126 to 148, or a base sequence in which u is t. (h) A polynucleotide consisting of a base sequence complementary to the base sequence shown in any of the sequence numbers 126 to 148 or the base sequence in which u is t, or a fragment of the polynucleotide containing 15 or more consecutive bases. (i) A polynucleotide containing a base sequence complementary to the base sequence shown in any of sequences 126–148 or a base sequence in which u is t, and (j) A polynucleotide that hybridizes with any of the polynucleotides in (f) to (i) under strict conditions.

14. The use according to any one of claims 8 to 13, wherein the apparatus is for determination by hybridization technique.

15. The use according to claim 14, wherein the hybridization technique is a nucleic acid array technique.

16. The use according to any one of claims 8 or 11, wherein the device comprises at least two nucleic acid probes capable of specifically binding to at least two polynucleotides selected from all biliary tract cancer markers described in claim 8 or 11 and / or at least two primers specifically recognizing and amplifying the polynucleotides.

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