Detection kit or device and detection method for biliary tract cancer

Abstract

The present 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, comprising a nucleic acid capable of specifically binding to miRNA in a specimen of a subject, and a method for detecting biliary tract cancer comprising in vitro measurement of the miRNA.

Description

[0001] This application is a divisional application of a patent application filed on June 11, 2015, with application number 201580030367.0 and invention name “Detection kit or device and detection method for biliary tract cancer”. Technical Field

[0002] The present invention relates to a kit or device for detecting biliary tract cancer, which is used to examine whether a subject has biliary tract cancer and contains a nucleic acid that can specifically bind to a specific miRNA, and a method for detecting biliary tract cancer, which comprises using the nucleic acid to measure the expression level of the miRNA. Background Art

[0003] The bile duct is the general excretory pathway from bile secretion by hepatocytes to its outflow into the duodenum. It is broadly divided into the intrahepatic bile duct within the liver and the extrahepatic bile duct outside the liver. The extrahepatic bile duct is broadly divided into three parts: the extrahepatic bile duct, which carries bile from the liver to the duodenum; the gallbladder, which temporarily stores and concentrates bile; and the duodenal papilla, or papilla, where the bile duct and main pancreatic duct open into the duodenum.

[0004] Most biliary tract cancers are caused by the cancerous transformation of the epithelial cells that line the bile duct lumen. Chemotherapy and radiation therapy are ineffective, and surgical resection based on early detection is the only curative treatment. However, biliary tract cancers initially present with no symptoms. Symptoms such as jaundice and itching develop initially as the cancer progresses, leading to bile duct obstruction and bile backflow into the blood vessels. Therefore, biliary tract cancer is often discovered while the cancer is advanced. Furthermore, intrahepatic biliary duct cancer rarely obstructs the extrahepatic bile duct, so most cases do not cause jaundice and the disease progresses asymptomatically. According to the 2011 statistics on cancer mortality rates by site in Japan published by the Cancer Control Information Center of the National Cancer Center, the number of deaths from biliary tract cancer rose to 18,186. From 2003 to 2005, the five-year relative survival rate for each site was 22.5% for men and 19.9% for women, second only to pancreatic cancer. The biliary tract is closely connected to vital organs such as the liver and pancreas, so metastasis to these organs can worsen the prognosis.

[0005] Biliary tract cancer is broadly divided into three types based on the site of occurrence: extrahepatic bile duct cancer, gallbladder cancer, and papillary cancer. Extrahepatic bile duct cancer is further divided into four types: the hilum at the entrance of the liver (hilar bile duct cancer), the upper part from the hilum to the gallbladder (upper bile duct cancer), the middle part from the gallbladder to the pancreas (middle bile duct cancer), and the lower part from the pancreas to the duodenal papilla (lower bile duct cancer). Cancers that originate in the bile duct closer to the liver are known to be more difficult to surgically treat and have a worse prognosis.

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

[0007] The initial diagnosis of biliary tract cancer usually uses low-invasive blood biochemical tests, tumor marker tests and abdominal ultrasound (non-patent literature 1). In the blood biochemical tests for detecting biliary tract cancer, for example, alkaline phosphatase, γ-GTP, bilirubin, etc. that rise due to liver dysfunction are used. Tumor markers for detecting biliary tract cancer are known, for example, CEA, CA19-9, DUPAN-2, CA195, CA242, IL-6, etc. As a method of using these tumor markers, when the concentration in the blood is higher or lower than a pre-set baseline value, it is suspected to be cancer. For example, as described in non-patent literature 2, when the baseline value of CEA is set to 5 ng / mL and the baseline value of CA19-9 is set to 37 U / mL, when a value above these values is displayed, it is suspected to be a cancer including 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 using the expression level of a protein in biliary tract tissue.

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

[0011] Prior art literature

[0012] Patent Literature

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

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

[0015] Non-patent literature

[0016] Non-patent document 1: Biliary tract cancer diagnosis and treatment guidelines compilation and publication committee, “Evidence-based biliary tract cancer diagnosis and treatment guidelines”, Medical Book Publishing Co., Ltd., 2007, p. 38-39

[0017] Non-patent document 2: Kiyoshi Kurokawa, Clinical Examination Data Handbook, 2013, p633, 636 Summary of the Invention

[0018] Problems to be solved by the invention

[0019] The present invention aims to discover a new tumor marker for biliary tract cancer and to provide a method for effectively detecting biliary tract cancer using a nucleic acid that can specifically bind to the marker. As described in Non-Patent Document 1, the initial diagnosis of biliary tract cancer generally uses minimally invasive blood biochemical tests, tumor marker tests, and abdominal ultrasound. The tumor detection rate (the probability of cancer being detected by imaging) of biliary tract cancer using abdominal ultrasound is 21 to 90% (Non-Patent Document 1), a wide range. The detection rate is particularly low when the cancer occupies the lower bile duct. In blood biochemical tests, for example, alkaline phosphatase, γ-GTP, and bilirubin, which are elevated due to liver dysfunction, are also used for biliary tract cancer detection. However, these blood biochemical tests cannot specifically detect biliary tract cancer. In addition, known tumor markers for biliary tract cancer detection include CEA, CA19-9, DUPAN-2, CA195, CA242, and IL-6. Among these, CEA is known to be elevated in 40% to 70% of patients with biliary tract cancer, and CA19-9 is known to be elevated in 50% to 79% of patients with biliary tract cancer (Non-Patent Document 1). However, these are not specific for biliary tract cancer, and Non-Patent Document 1 also states that they are difficult to use for early diagnosis. Furthermore, Non-Patent Document 1 states that the clinical usefulness of DUPAN-2, CA195, CA242, and IL-6 is unclear. Therefore, it is also believed that the use of conventional tumor markers may lead to the misdiagnosis of other cancers and / or benign tumors and / or benign diseases of the biliary tract and / or organs surrounding the biliary tract.

[0020] In addition, although it is still in the research stage, there are reports such as the following that use 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 protein expression levels in biliary tract tissue. However, this method requires surgical tissue resection to obtain a specimen, a procedure that places a heavy physical burden on the patient and is therefore not a suitable diagnostic method. Furthermore, Patent Document 1 does not describe the method's specific accuracy, sensitivity, and specificity for identifying biliary tract cancer, making it lacking industrial practicality.

[0022] Patent Document 2 describes a method for diagnosing digestive tract cancers, including biliary tract cancer, using mRNA genes extracted from cells in the blood (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 when developing it for actual testing. Furthermore, mRNA is easily degraded in blood and is unstable, making it unsuitable for testing.

[0023] As such, existing tumor markers for biliary tract cancer detection have low performance, or for markers in the research phase, performance and detection methods are not specifically demonstrated. Consequently, their use can lead to unnecessary additional testing due to misidentification of healthy individuals as biliary tract cancer patients, or to missed biliary tract cancer patients, resulting in lost treatment opportunities. Furthermore, measuring dozens to hundreds of genes increases testing costs, making large-scale screening, such as health checks, difficult to implement. Furthermore, collecting biliary tract tissue for tumor marker measurement is highly invasive and undesirable. Therefore, there is a need for biliary tract cancer markers that can be detected from blood, which can be collected minimally invasively, and that can accurately distinguish biliary tract cancer patients from biliary tract cancer patients and healthy individuals from healthy individuals. In particular, since early detection and resection are the only curative treatment for biliary tract cancer, highly sensitive biliary tract cancer markers are highly desired.

[0024] Methods for solving problems

[0025] The present inventors conducted in-depth research to solve the above-mentioned problems, and as a result, they found multiple genes that can be used as detection markers for biliary tract cancer from blood that can be collected with low invasiveness. They also found that by using nucleic acids that can specifically bind to them, biliary tract cancer can be significantly detected, thus completing the present invention.

[0026] <Overview of the Invention>

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

[0028] (1) Kit for detecting biliary tract cancer, which contains a substance that can bind 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 A nucleic acid that specifically binds to at least one of the following polynucleotides: miR-656, 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) The kit according to (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, and miR-6789-5p is hsa-miR-6789-5p. 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-615 615-5p, 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, and miR-466 is hsa-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, and miR-7704 is hsa-miR-7705. 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 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, and miR-24-3p is hsa-miR-24-3p. -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-miR-6768-5p 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, and 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-miR4746-3p,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-7600 is hsa-miR-7601 for hsa-miR-760, miR-3162-5p for hsa-miR-3162-5p, miR-3178 for hsa-miR-3178, miR-940 for hsa-miR-940, miR-4271 for hsa-miR-4271, miR-6769b-5p for hsa-miR-6769b-5p, miR-4508 for hsa-miR-4508, miR-6826-5p for hsa-miR-6826-5p, miR-6757-5p for hsa-miR-6757-5p, miR-3131 for hsa-miR-3131, and miR-1343-3p for hsa-miR-1343-3p. ,

[0030] (3) The kit according to (1) or (2), wherein the nucleic acid is a polynucleotide selected from the polynucleotides shown in the following (a) to (e),

[0031] (a) a polynucleotide consisting of the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0032] (b) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478,

[0033] (c) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0034] (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, and

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

[0036] (4) The kit according to any one of (1) to (3), further comprising a kit capable of interacting 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 A nucleic acid that specifically binds to at least one of the polynucleotides selected from the group consisting of 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) The kit according to (4) or (5), wherein the nucleic acid is a polynucleotide selected from the polynucleotides shown in the following (f) to (j),

[0039] (f) a polynucleotide consisting of the base sequence represented by any of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0040] (g) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 126 to 148,

[0041] (h) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0042] (i) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, and

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

[0044] (7) The kit according to any one of (1) to (6), wherein the kit contains at least two or more nucleic acids that can specifically bind to at least two or more 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-4449、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 A nucleic acid that specifically binds to at least one of the following polynucleotides: miR-656, 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 device 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, and 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, and miR-6789-5p is hsa-miR-6789-5p. 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, and miR-615-5p is hsa-miR-615-5p. 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, and miR-466 is hsa-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, and miR-7704 is hsa-miR-7705. 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 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, and miR-24-3p is hsa-miR-24-3p. -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-miR-6768-5p 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, and 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-miR4746-3p,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-7600 is hsa-miR-7601 for hsa-miR-760, miR-3162-5p for hsa-miR-3162-5p, miR-3178 for hsa-miR-3178, miR-940 for hsa-miR-940, miR-4271 for hsa-miR-4271, miR-6769b-5p for hsa-miR-6769b-5p, miR-4508 for hsa-miR-4508, miR-6826-5p for hsa-miR-6826-5p, miR-6757-5p for hsa-miR-6757-5p, miR-3131 for hsa-miR-3131, and miR-1343-3p for hsa-miR-1343-3p. ,

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

[0048] (a) a polynucleotide consisting of the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0049] (b) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478,

[0050] (c) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0051] (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, and

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

[0053] (11) The device according to any one of (8) to (10), further comprising a device capable of interacting with other biliary 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, and miR-6774-5p. A nucleic acid that specifically binds to at least one of the polynucleotides selected from the group consisting of 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 device 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-1233-5 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, 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.

[0055] (13) The device according to (11) or (12), wherein the nucleic acid is a polynucleotide selected from the polynucleotides shown in the following (f) to (j),

[0056] (f) a polynucleotide consisting of the base sequence represented by any of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0057] (g) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 126 to 148,

[0058] (h) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0059] (i) A polynucleotide comprising a base sequence complementary to the base sequence shown in any one of SEQ ID NOs: 126 to 148 or a base sequence in which u is t in the base sequence, and

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

[0061] (14) The apparatus according to any one of (8) to (13), wherein the apparatus is an apparatus for measurement by a hybridization technique.

[0062] (15) The apparatus according to (14), wherein the hybridization technique is a nucleic acid array technique.

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

[0064] (17) A method for detecting biliary tract cancer, comprising: using the kit according to any one of (1) to (7) or the apparatus according to any one of (8) to (16) to measure the expression level of a target nucleic acid in a specimen of a subject, and using the measured expression level and the control expression level of a healthy subject measured in the same manner to perform an in vitro evaluation of whether the subject has biliary tract cancer or not.

[0065] (18) The method according to (17), wherein the subject is a human.

[0066] (19) The method according to (17) or (18), wherein the specimen is blood, serum or plasma.

[0067] <Definition 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 formed in the biliary tract. Specifically, it includes extrahepatic cholangiocarcinoma, gallbladder cancer, papillary cancer, duodenal papillary cancer, intrahepatic cholangiocarcinoma, etc.

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

[0071] The representations of abbreviations such as nucleotide, polynucleotide, DNA, RNA, etc. follow the "Guidelines for the Preparation of Specifications Containing Base Sequences or Amino Acid Sequences" (edited by the Japan Patent Office) and the common practice in this technical field.

[0072] In this specification, the term "polynucleotide" refers to nucleic acids including any one of RNA, DNA, and RNA / DNA (chimera). In addition, the above-mentioned DNA includes any one of cDNA, genomic DNA, and synthetic DNA. In addition, the above-mentioned RNA includes any one 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 a specified base sequence (which can be any one of a natural sequence or a non-natural sequence) using, for example, an automatic nucleic acid synthesizer. In this specification, "non-natural sequence" is intended to be used in a broad sense, including sequences that differ from natural sequences, such as sequences that include substitutions, deletions, insertions, and / or additions of one or more nucleotides (i.e., mutant sequences), sequences that include one or more modified nucleotides (i.e., modified sequences), etc. In addition, in this specification, polynucleotides and nucleic acids are used interchangeably.

[0073] In this specification, a "fragment" is a polynucleotide having a continuous portion of the base sequence of a polynucleotide, and desirably has a length of 15 bases or more, preferably 17 bases or more, and more preferably 19 bases or more.

[0074] In this specification, the term "gene" is intended to include not only RNA and double-stranded DNA but also single-stranded DNA such as the positive strand (or sense strand) or complementary strand (or antisense strand) constituting them. Furthermore, the length thereof is not particularly limited.

[0075] Therefore, in this specification, "gene", unless otherwise specified, includes: double-stranded DNA comprising human genomic DNA, single-stranded DNA (positive strand), single-stranded DNA (complementary strand) comprising cDNA having a sequence complementary to the positive strand, microRNA (miRNA), and any of their fragments and transcription products. In addition, the "gene" is not only the "gene" shown by a specific base sequence (or sequence number), but also includes RNA with the same biological function as the RNA encoded by them, such as "nucleic acids" encoding homologs (i.e., homologs or orthologs), mutants such as gene polymorphisms, and derivatives. As such "nucleic acids" encoding homologs, mutants or derivatives, specifically, "nucleic acids" having a base sequence that hybridizes with the base sequence shown in any one of SEQ ID NOs. 1 to 509 or the complementary sequence of the base sequence in which u is t in the base sequence under the stringent conditions described below can be cited. In addition, "gene" can include, for example, expression control regions, coding regions, exons or introns, regardless of the difference in functional regions. Furthermore, the "gene" may be contained in cells, released outside the cells and exist independently, or may be enclosed in small vesicles called exosomes.

[0076] As used herein, "exosomes" are small vesicles surrounded by a lipid bilayer secreted by cells. Exosomes originate from multivesicular endosomes and, when released into the extracellular environment, may contain biological substances such as "genes" such as RNA and DNA, as well as proteins. Exosomes are known to be present in body fluids such as blood, serum, plasma, serum, and lymph.

[0077] As used herein, "transcript" refers to RNA synthesized using the DNA sequence of a gene as a template. RNA polymerase binds to a region upstream of the gene called the promoter, where it binds ribonucleotides complementary to the DNA base sequence at the 3' end to synthesize RNA. This RNA contains not only the gene itself but also the entire sequence from the transcription start point to the end of the poly A sequence, represented by expression control regions, coding regions, exons, or introns.

[0078] In addition, as used herein, "microRNA (miRNA)" is intended to be used as a non-coding RNA of 15 to 25 bases, unless otherwise specified, that is transcribed as a hairpin-like RNA precursor, cleaved by a dsRNA cleavage enzyme with RNase III cleavage activity, and incorporated into a protein complex called RISC, which participates in the translational repression of mRNA. Furthermore, the "miRNA" used herein includes not only "miRNAs" represented by specific base sequences (or sequence numbers), but also precursors (pre-miRNA, pri-miRNA) of such "miRNAs," as well as miRNAs with equivalent biological functions, such as homologs (i.e., homologs or orthologs), mutants due to genetic 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 that hybridize under stringent conditions described below to the complementary sequence of any of the specific base sequences represented by any of SEQ ID NOs. 1 to 509. In addition, the "miRNA" used in this specification can be a gene product of a miR gene, such a gene product comprising a mature miRNA (for example, a non-coding RNA of 15 to 25 bases or 19 to 25 bases involved in translational inhibition of mRNA as described above) or a miRNA precursor (for example, a pre-miRNA or pri-miRNA as described above).

[0079] The term "probe" herein includes a polynucleotide for specifically detecting RNA produced by gene expression or a polynucleotide derived from the RNA and / or a polynucleotide complementary thereto.

[0080] The term "primer" as used herein includes a polynucleotide that specifically identifies and amplifies RNA produced by gene expression or a polynucleotide derived from the RNA and / or a polynucleotide complementary thereto.

[0081] Here, a complementary polynucleotide (complementary strand, minus strand) refers to a polynucleotide that is complementary to the full-length sequence of a polynucleotide consisting of a base sequence defined by any of SEQ ID NOs: 1 to 509, or a base sequence in which u is t in such a base sequence, or a partial sequence thereof (herein referred to as the plus strand for convenience), based on base pair relationships such as A:T(U) and G:C. However, such a complementary strand is not limited to being completely complementary to the base sequence of the target plus strand and may also be complementary to the extent that it can hybridize with the target plus strand under stringent conditions.

[0082] As used herein, "stringent conditions" refer to conditions under which a nucleic acid probe hybridizes to its target sequence to a greater degree (e.g., a value greater than the mean of background values + the standard error of background values × 2) than it hybridizes to other sequences. Stringent conditions are sequence-dependent and vary depending on the environment in which hybridization is performed. 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 below.

[0083] The "Tm value" herein refers to the temperature at which the double-stranded portion of a polynucleotide denatures into a single strand and the double strands and single strands exist in a 1:1 ratio.

[0084] As used herein, "mutant" refers to a natural mutant caused by polymorphism, mutation, etc. in the case of nucleic acids, or a mutant comprising a deletion, substitution, addition, or insertion of one or more bases in any of the base sequences of SEQ ID NOs. 1 to 509, or a base sequence in which u is t in the base sequence, or a partial sequence thereof, or a mutant showing a % identity of about 90% or more, about 95% or more, about 97% or more, about 98% or more, or about 99% or more with the respective base sequences or partial sequences thereof, or a nucleic acid that hybridizes with a polynucleotide or oligonucleotide comprising the base sequence or a partial sequence thereof under the stringent conditions defined above.

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

[0086] In this specification, mutants can be prepared using known techniques such as site-directed mutagenesis and mutation introduction using PCR.

[0087] As used herein, "% identity" can be determined using the above-mentioned protein or gene search systems utilizing BLAST or FASTA, with or without introducing gaps (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 a modified nucleic acid, and includes, but is not limited to, derivatives labeled with fluorophores, etc., derivatives containing modified nucleotides (for example, nucleotides containing groups such as halogens, alkyl groups such as methyl groups, alkoxy groups such as methoxy groups, thio groups, carboxymethyl groups, and nucleotides that have undergone base reconstruction, double bond saturation, deamination, replacement of oxygen molecules with sulfur molecules, etc.), PNA (peptide nucleic acid; Nielsen, PE et al., 1991, Science, Vol. 254, p1497-500), LNA (locked nucleic acid; Obika, S. et al., 1998, Tetrahedron Lett., Vol. 39, p5401-5404), etc.

[0089] In this specification, the "nucleic acid" that can specifically bind to a polynucleotide selected from the above-mentioned miRNA group as a biliary tract cancer marker is a synthetic or modulated nucleic acid, specifically, a "nucleic acid probe" or a "primer", which is used directly or indirectly to detect the presence of biliary tract cancer in a subject, or to diagnose the presence of biliary tract cancer, the extent of biliary tract cancer, the improvement of biliary tract cancer, the degree of improvement, the sensitivity to biliary tract cancer treatment, or to screen for candidate substances useful for the prevention, improvement or treatment of biliary tract cancer. They include nucleotides, oligonucleotides and polynucleotides that can specifically identify and bind to the transcripts shown in any one of sequence numbers 1 to 509 or their cDNA synthesized nucleic acids in a specimen such as blood, urine, etc. in a living body, especially in a body fluid such as blood. Based on the above properties, these nucleotides, oligonucleotides and polynucleotides can be effectively used as probes for detecting the above-mentioned genes expressed in a living body, tissue, cell, etc., and can also be effectively used as primers for amplifying the above-mentioned genes expressed in a living body.

[0090] The term "detection" used in this specification can be replaced with the term "inspection," "measurement," "detection," or "determination support." Furthermore, the term "evaluation" used in this specification includes supporting diagnosis or evaluation based on the results of an inspection or measurement.

[0091] As used herein, the term "subject" refers to mammals, including humans, primates such as chimpanzees, pets such as dogs and cats, livestock such as cattle, horses, sheep, and goats, and rodents such as mice and rats. Furthermore, the term "healthy subject" also refers to such mammals, meaning animals that do not have the cancer being tested.

[0092] As used herein, "P" or "P-value" indicates the probability of observing an extreme statistic compared to the statistic actually calculated from the data under the null hypothesis in a statistical test. Therefore, a smaller "P" or "P-value" indicates a significant difference between the comparison subjects.

[0093] In this specification, "sensitivity" refers to the value of (number of true positives) / (number of true positives + number of false negatives). If the sensitivity is high, biliary tract cancer can be detected early, leading to complete resection of the cancer and a reduction in the recurrence rate.

[0094] In this specification, "specificity" refers to (the number of true negatives) / (the number of true negatives + the number of false positives). If the specificity is high, it can prevent the implementation of unnecessary additional examinations caused by misidentifying healthy people as patients with biliary tract cancer, thereby reducing the burden on patients and medical expenses.

[0095] In this specification, "accuracy" refers to the value of (number of true positives + number of true negatives) / (total number of cases). Accuracy indicates the proportion of correct discrimination results for all samples and is the primary indicator for evaluating detection performance.

[0096] The "specimen" used in this specification as the subject of determination, detection, or diagnosis refers to tissues and biological materials whose gene expression changes with the development and progression of biliary tract cancer and the effectiveness of treatment for biliary tract cancer. Specifically, it refers to biliary tissue and its surrounding blood vessels, lymph nodes, and organs. It also refers to suspected metastatic organs, skin, and body fluids such as blood, urine, saliva, sweat, and tissue leaching fluid, serum and plasma prepared from blood, as well as feces and hair. It further refers to biological samples extracted from these, specifically, genes such as RNA and miRNA.

[0097] As used herein, the term "hsa-miR-125a-3p gene" or "hsa-miR-125a-3p" includes the hsa-miR-125a-3p gene described in SEQ ID NO: 1 (miRBase Accession No. MIMAT0004602) and homologs or orthologs in other biological species. The hsa-miR-125a-3p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12, pp. 735-739. Furthermore, as a precursor of "hsa-miR-125a-3p", "hsa-mir-125a" (miRBase Accession No. MI0000469, SEQ ID NO: 149) is known to have a hairpin-like structure.

[0098] As used herein, the term “hsa-miR-6893-5p gene” or “hsa-miR-6893-5p” includes the hsa-miR-6893-5p gene described in SEQ ID NO: 2 (miRBase Accession No. MIMAT0027686) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6893-5p,” “hsa-mir-6893” (miRBase Accession No. MI0022740, SEQ ID NO: 150) is known to have a hairpin-like structure.

[0099] As used herein, the term "hsa-miR-204-3p gene" or "hsa-miR-204-3p" includes the hsa-miR-204-3p gene described in SEQ ID NO: 3 (miRBase Accession No. MIMAT0022693) and homologs or orthologs in other species. The hsa-miR-204-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. Furthermore, as a precursor of "hsa-miR-204-3p", "hsa-mir-204" (miRBase Accession No. MI0000284, SEQ ID NO: 151) is known to have a hairpin-like structure.

[0100] As used herein, the term "hsa-miR-4476 gene" or "hsa-miR-4476" includes the hsa-miR-4476 gene described in SEQ ID NO: 4 (miRBase Accession No. MIMAT0019003) and homologs or orthologs in other species. 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 No. MI0016828, SEQ ID NO: 152) is known to have a hairpin-like structure.

[0101] As used herein, the term “hsa-miR-4294 gene” or “hsa-miR-4294” includes the hsa-miR-4294 gene described in SEQ ID NO: 5 (miRBase Accession No. MIMAT0016849) and homologs or orthologs in other species. The hsa-miR-4294 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-4294”, “hsa-mir-4294” (miRBase Accession No. MI0015827, SEQ ID NO: 153) is known to have a hairpin-like structure.

[0102] As used herein, the term "hsa-miR-150-3p gene" or "hsa-miR-150-3p" includes the hsa-miR-150-3p gene described in SEQ ID NO: 6 (miRBase Accession No. MIMAT0004610) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-150-3p", "hsa-mir-150" (miRBase Accession No. MI0000479, SEQ ID NO: 154) is known to have a hairpin-like structure.

[0103] As used herein, the term "hsa-miR-6729-5p gene" or "hsa-miR-6729-5p" includes the hsa-miR-6729-5p gene described in SEQ ID NO: 7 (miRBase Accession No. MIMAT0027359) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6729-5p", "hsa-mir-6729" (miRBase Accession No. MI0022574, SEQ ID NO: 155) is known to have a hairpin-like structure.

[0104] As used herein, the term “hsa-miR-7641 gene” or “hsa-miR-7641” includes the hsa-miR-7641 gene described in SEQ ID NO: 8 (miRBase Accession No. MIMAT0029782) and homologs or orthologs of other biological species. 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 of “hsa-miR-7641”, “hsa-mir-7641-1” and “hsa-mir-7641-2” (miRBase Accession Nos. MI0024975, MI0024976, SEQ ID NOs: 156, 157) are known as hairpin-like structure precursors.

[0105] As used herein, the term "hsa-miR-6765-3p gene" or "hsa-miR-6765-3p" includes the hsa-miR-6765-3p gene described in SEQ ID NO: 9 (miRBase Accession No. MIMAT0027431) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6765-3p", "hsa-mir-6765" (miRBase Accession No. MI0022610, SEQ ID NO: 158) is known to have a hairpin-like structure.

[0106] As used herein, the term "hsa-miR-6820-5p gene" or "hsa-miR-6820-5p" includes the hsa-miR-6820-5p gene described in SEQ ID NO: 10 (miRBase Accession No. MIMAT0027540) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6820-5p", "hsa-mir-6820" (miRBase Accession No. MI0022665, SEQ ID NO: 159) is known to have a hairpin-like structure.

[0107] As used herein, the term "hsa-miR-575 gene" or "hsa-miR-575" includes the hsa-miR-575 gene described in SEQ ID NO: 11 (miRBase Accession No. MIMAT0003240) and homologs or orthologs in other species. 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 of "hsa-miR-575," "hsa-mir-575" (miRBase Accession No. MI0003582, SEQ ID NO: 160) is known to have a hairpin-like structure.

[0108] As used herein, the term "hsa-miR-6836-3p gene" or "hsa-miR-6836-3p" includes the hsa-miR-6836-3p gene described in SEQ ID NO: 12 (miRBase Accession No. MIMAT0027575) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6836-3p", "hsa-mir-6836" (miRBase Accession No. MI0022682, SEQ ID NO: 161) is known to have a hairpin-like structure.

[0109] As used herein, the term “hsa-miR-1469 gene” or “hsa-miR-1469” includes the hsa-miR-1469 gene described in SEQ ID NO: 13 (miRBase Accession No. MIMAT0007347) and homologs or orthologs in other biological species. The hsa-miR-1469 gene can be obtained by the method described in Kawaji H et al., 2008, BMC Genomics, Vol. 9, p157. In addition, as a precursor of “hsa-miR-1469”, “hsa-mir-1469” (miRBase Accession No. MI0007074, SEQ ID NO: 162) that forms a hairpin-like structure is known.

[0110] As used herein, the term "hsa-miR-663a gene" or "hsa-miR-663a" includes the hsa-miR-663a gene described in SEQ ID NO: 14 (miRBase Accession No. MIMAT0003326) and homologs or orthologs in other species. The hsa-miR-663a gene can be obtained by the method described in Cummins JM et al., 2006, Proc Natl Acad Sci USA, Vol. 103, p3687-3692. In addition, as a precursor of "hsa-miR-663a", "hsa-mir-663a" (miRBase Accession No. MI0003672, SEQ ID NO: 163) is known to have a hairpin-like structure.

[0111] As used herein, the term “hsa-miR-6075 gene” or “hsa-miR-6075” includes the hsa-miR-6075 gene described in SEQ ID NO: 15 (miRBase Accession No. MIMAT0023700) and homologs or orthologs in other species. The hsa-miR-6075 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor of “hsa-miR-6075,” “hsa-mir-6075” (miRBase Accession No. MI0020352, SEQ ID NO: 164) is known to have a hairpin-like structure.

[0112] As used herein, the term "hsa-miR-4634 gene" or "hsa-miR-4634" includes the hsa-miR-4634 gene described in SEQ ID NO: 16 (miRBase Accession No. MIMAT0019691) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4634," "hsa-mir-4634" (miRBase Accession No. MI0017261, SEQ ID NO: 165) is known to have a hairpin-like structure.

[0113] As used herein, the term “hsa-miR-423-5p gene” or “hsa-miR-423-5p” includes the hsa-miR-423-5p gene described in SEQ ID NO: 17 (miRBase Accession No. MIMAT0004748) and homologs or orthologs in other biological species. 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 No. MI0001445, SEQ ID NO: 166) is known to have a hairpin-like structure.

[0114] As used herein, the term "hsa-miR-4454 gene" or "hsa-miR-4454" includes the hsa-miR-4454 gene described in SEQ ID NO: 18 (miRBase Accession No. MIMAT0018976) and homologs or orthologs in other species. 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 No. MI0016800, SEQ ID NO: 167) is known to have a hairpin-like structure.

[0115] As used herein, the term "hsa-miR-7109-5p gene" or "hsa-miR-7109-5p" includes the hsa-miR-7109-5p gene described in SEQ ID NO: 19 (miRBase Accession No. MIMAT0028115) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-7109-5p", "hsa-mir-7109" (miRBase Accession No. MI0022960, SEQ ID NO: 168) is known to have a hairpin-like structure.

[0116] As used herein, the term “hsa-miR-6789-5p gene” or “hsa-miR-6789-5p” includes the hsa-miR-6789-5p gene described in SEQ ID NO: 20 (miRBase Accession No. MIMAT0027478) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6789-5p,” “hsa-mir-6789” (miRBase Accession No. MI0022634, SEQ ID NO: 169) is known to have a hairpin-like structure.

[0117] As used herein, the term “hsa-miR-6877-5p gene” or “hsa-miR-6877-5p” includes the hsa-miR-6877-5p gene described in SEQ ID NO: 21 (miRBase Accession No. MIMAT0027654) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6877-5p,” “hsa-mir-6877” (miRBase Accession No. MI0022724, SEQ ID NO: 170) is known to have a hairpin-like structure.

[0118] As used herein, the term “hsa-miR-4792 gene” or “hsa-miR-4792” includes the hsa-miR-4792 gene described in SEQ ID NO: 22 (miRBase Accession No. MIMAT0019964) and homologs or orthologs in other species. 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 No. MI0017439, SEQ ID NO: 171) is known to have a hairpin-like structure.

[0119] As used herein, the term "hsa-miR-4530 gene" or "hsa-miR-4530" includes the hsa-miR-4530 gene described in SEQ ID NO: 23 (miRBase Accession No. MIMAT0019069) and homologs or orthologs in other species. The hsa-miR-4530 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4530", "hsa-mir-4530" (miRBase Accession No. MI0016897, SEQ ID NO: 172) is known to have a hairpin-like structure.

[0120] As used herein, the term "hsa-miR-7975 gene" or "hsa-miR-7975" includes the hsa-miR-7975 gene described in SEQ ID NO: 24 (miRBase Accession No. MIMAT0031178) and homologs or orthologs in other biological species. The hsa-miR-7975 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online version. In addition, as a precursor of "hsa-miR-7975", "hsa-mir-7975" (miRBase Accession No. MI0025751, SEQ ID NO: 173) is known to have a hairpin-like structure.

[0121] As used herein, the term "hsa-miR-6724-5p gene" or "hsa-miR-6724-5p" includes the hsa-miR-6724-5p gene described in SEQ ID NO: 25 (miRBase Accession No. MIMAT0025856) and homologs or orthologs in other species. 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 of "hsa-miR-6724-5p," "hsa-mir-6724" (miRBase Accession No. MI0022559, SEQ ID NO: 174) is known to have a hairpin-like structure.

[0122] As used herein, the term “hsa-miR-8073 gene” or “hsa-miR-8073” includes the hsa-miR-8073 gene described in SEQ ID NO: 26 (miRBase Accession No. MIMAT0031000) and homologs or orthologs in other species. The hsa-miR-8073 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. In addition, as a precursor of “hsa-miR-8073”, “hsa-mir-8073” (miRBase Accession No. MI0025909, SEQ ID NO: 175) is known to have a hairpin-like structure.

[0123] As used herein, the term “hsa-miR-7977 gene” or “hsa-miR-7977” includes the hsa-miR-7977 gene described in SEQ ID NO: 27 (miRBase Accession No. MIMAT0031180) and homologs or orthologs in other biological species. The hsa-miR-7977 gene can be obtained by the method described in Velthut-Meikas A et al., 2013, Mol Endocrinol, online version. In addition, as a precursor of “hsa-miR-7977”, “hsa-mir-7977” (miRBase Accession No. MI0025753, SEQ ID NO: 176) is known to form a hairpin-like structure.

[0124] As used herein, the term "hsa-miR-1231 gene" or "hsa-miR-1231" includes the hsa-miR-1231 gene described in SEQ ID NO: 28 (miRBase Accession No. MIMAT0005586) and homologs or orthologs in other species. The hsa-miR-1231 gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor of "hsa-miR-1231," "hsa-mir-1231" (miRBase Accession No. MI0006321, SEQ ID NO: 177) is known to have a hairpin-like structure.

[0125] As used herein, the term “hsa-miR-6799-5p gene” or “hsa-miR-6799-5p” includes the hsa-miR-6799-5p gene described in SEQ ID NO: 29 (miRBase Accession No. MIMAT0027498) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6799-5p,” “hsa-mir-6799” (miRBase Accession No. MI0022644, SEQ ID NO: 178) is known to have a hairpin-like structure.

[0126] As used herein, the term "hsa-miR-615-5p gene" or "hsa-miR-615-5p" includes the hsa-miR-615-5p gene described in SEQ ID NO: 30 (miRBase Accession No. MIMAT0004804) and homologs or orthologs in other species. The hsa-miR-615-5p 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 of "hsa-miR-615-5p", "hsa-mir-615" (miRBase Accession No. MI0003628, SEQ ID NO: 179) is known to have a hairpin-like structure.

[0127] As used herein, the term "hsa-miR-4450 gene" or "hsa-miR-4450" includes the hsa-miR-4450 gene described in SEQ ID NO: 31 (miRBase Accession No. MIMAT0018971) and homologs or orthologs in other species. 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 No. MI0016795, SEQ ID NO: 180) is known to have a hairpin-like structure.

[0128] As used herein, the term "hsa-miR-6726-5p gene" or "hsa-miR-6726-5p" includes the hsa-miR-6726-5p gene described in SEQ ID NO: 32 (miRBase Accession No. MIMAT0027353) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6726-5p", "hsa-mir-6726" (miRBase Accession No. MI0022571, SEQ ID NO: 181) is known to have a hairpin-like structure.

[0129] As used herein, the term “hsa-miR-6875-5p gene” or “hsa-miR-6875-5p” includes the hsa-miR-6875-5p gene described in SEQ ID NO: 33 (miRBase Accession No. MIMAT0027650) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6875-5p,” “hsa-mir-6875” (miRBase Accession No. MI0022722, SEQ ID NO: 182) is known to have a hairpin-like structure.

[0130] As used herein, the term "hsa-miR-4734 gene" or "hsa-miR-4734" includes the hsa-miR-4734 gene described in SEQ ID NO: 34 (miRBase Accession No. MIMAT0019859) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4734," "hsa-mir-4734" (miRBase Accession No. MI0017371, SEQ ID NO: 183) is known to have a hairpin-like structure.

[0131] As used herein, the term "hsa-miR-16-5p gene" or "hsa-miR-16-5p" includes the hsa-miR-16-5p gene described in SEQ ID NO: 35 (miRBase Accession No. MIMAT0000069) and homologs or orthologs in other biological species. 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. Furthermore, as precursors of "hsa-miR-16-5p", "hsa-mir-16-1" and "hsa-mir-16-2" (miRBase Accession Nos. MI0000070 and MI0000115, SEQ ID NOs. 184 and 185) are known.

[0132] As used herein, the term "hsa-miR-602 gene" or "hsa-miR-602" includes the hsa-miR-602 gene described in SEQ ID NO: 36 (miRBase Accession No. MIMAT0003270) and homologs or orthologs in other species. 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, p3687-3692. In addition, as a precursor of "hsa-miR-602", "hsa-mir-602" (miRBase Accession No. MI0003615, SEQ ID NO: 186) is known to have a hairpin-like structure.

[0133] As used herein, the term "hsa-miR-4651 gene" or "hsa-miR-4651" includes the hsa-miR-4651 gene described in SEQ ID NO: 37 (miRBase Accession No. MIMAT0019715) and homologs or orthologs in other species. The hsa-miR-4651 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 of "hsa-miR-4651," "hsa-mir-4651" (miRBase Accession No. MI0017279, SEQ ID NO: 187) is known to have a hairpin-like structure.

[0134] As used herein, the term “hsa-miR-8069 gene” or “hsa-miR-8069” includes the hsa-miR-8069 gene described in SEQ ID NO: 38 (miRBase Accession No. MIMAT0030996) and homologs or orthologs in other species. The hsa-miR-8069 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. In addition, as a precursor of “hsa-miR-8069”, “hsa-mir-8069” (miRBase Accession No. MI0025905, SEQ ID NO: 188) is known to have a hairpin-like structure.

[0135] As used herein, the term "hsa-miR-1238-5p gene" or "hsa-miR-1238-5p" includes the hsa-miR-1238-5p gene described in SEQ ID NO: 39 (miRBase Accession No. MIMAT0022947) and homologs or orthologs in other biological species. The hsa-miR-1238-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor of "hsa-miR-1238-5p", "hsa-mir-1238" (miRBase Accession No. MI0006328, SEQ ID NO: 189) is known to have a hairpin-like structure.

[0136] As used herein, the term "hsa-miR-6880-5p gene" or "hsa-miR-6880-5p" includes the hsa-miR-6880-5p gene described in SEQ ID NO: 40 (miRBase Accession No. MIMAT0027660) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6880-5p", "hsa-mir-6880" (miRBase Accession No. MI0022727, SEQ ID NO: 190) is known to have a hairpin-like structure.

[0137] As used herein, the term “hsa-miR-8072 gene” or “hsa-miR-8072” includes the hsa-miR-8072 gene described in SEQ ID NO: 41 (miRBase Accession No. MIMAT0030999) and homologs or orthologs in other species. The hsa-miR-8072 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. In addition, as a precursor of “hsa-miR-8072”, “hsa-mir-8072” (miRBase Accession No. MI0025908, SEQ ID NO: 191) is known to have a hairpin-like structure.

[0138] As used herein, the term “hsa-miR-4723-5p gene” or “hsa-miR-4723-5p” includes the hsa-miR-4723-5p gene described in SEQ ID NO: 42 (miRBase Accession No. MIMAT0019838) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-4723-5p,” “hsa-mir-4723” (miRBase Accession No. MI0017359, SEQ ID NO: 192) is known to have a hairpin-like structure.

[0139] As used herein, the term "hsa-miR-4732-5p gene" or "hsa-miR-4732-5p" includes the hsa-miR-4732-5p gene described in SEQ ID NO: 43 (miRBase Accession No. MIMAT0019855) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4732-5p", "hsa-mir-4732" (miRBase Accession No. MI0017369, SEQ ID NO: 193) is known to have a hairpin-like structure.

[0140] As used herein, the term “hsa-miR-6125 gene” or “hsa-miR-6125” includes the hsa-miR-6125 gene described in SEQ ID NO: 44 (miRBase Accession No. MIMAT0024598) and homologs or orthologs in other species. 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 of “hsa-miR-6125,” “hsa-mir-6125” (miRBase Accession No. MI0021259, SEQ ID NO: 194) is known to have a hairpin-like structure.

[0141] As used herein, the term "hsa-miR-6090 gene" or "hsa-miR-6090" includes the hsa-miR-6090 gene described in SEQ ID NO: 45 (miRBase Accession No. MIMAT0023715) and homologs or orthologs in other biological species. The hsa-miR-6090 gene can be obtained by the method described in Yoo JK et al., 2012, Stem Cells Dev, Vol. 21, pp. 2049-2057. Furthermore, as a precursor of "hsa-miR-6090", "hsa-mir-6090" (miRBase Accession No. MI0020367, SEQ ID NO: 195) is known to have a hairpin-like structure.

[0142] As used herein, the term "hsa-miR-7114-5p gene" or "hsa-miR-7114-5p" includes the hsa-miR-7114-5p gene described in SEQ ID NO: 46 (miRBase Accession No. MIMAT0028125) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-7114-5p", "hsa-mir-7114" (miRBase Accession No. MI0022965, SEQ ID NO: 196) is known to have a hairpin-like structure.

[0143] As used herein, the term "hsa-miR-564 gene" or "hsa-miR-564" includes the hsa-miR-564 gene described in SEQ ID NO: 47 (miRBase Accession No. MIMAT0003228) and homologs or orthologs in other species. 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. In addition, as a precursor of "hsa-miR-564", "hsa-mir-564" (miRBase Accession No. MI0003570, SEQ ID NO: 197) is known to have a hairpin-like structure.

[0144] As used herein, the term "hsa-miR-451a gene" or "hsa-miR-451a" includes the hsa-miR-451a gene described in SEQ ID NO: 48 (miRBase Accession No. MIMAT0001631) and homologs or orthologs in other species. 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. In addition, as a precursor of "hsa-miR-451a", "hsa-mir-451a" (miRBase Accession No. MI0001729, SEQ ID NO: 198) is known to have a hairpin-like structure.

[0145] As used herein, the term "hsa-miR-3135b gene" or "hsa-miR-3135b" includes the hsa-miR-3135b gene described in SEQ ID NO: 49 (miRBase Accession No. MIMAT0018985) and homologs or orthologs in other species. The hsa-miR-3135b gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-3135b", "hsa-mir-3135b" (miRBase Accession No. MI0016809, SEQ ID NO: 199) is known to have a hairpin-like structure.

[0146] As used herein, the term "hsa-miR-4497 gene" or "hsa-miR-4497" includes the hsa-miR-4497 gene described in SEQ ID NO: 50 (miRBase Accession No. MIMAT0019032) and homologs or orthologs in other species. 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 No. MI0016859, SEQ ID NO: 200) is known to have a hairpin-like structure.

[0147] As used herein, the term "hsa-miR-4665-5p gene" or "hsa-miR-4665-5p" includes the hsa-miR-4665-5p gene described in SEQ ID NO: 51 (miRBase Accession No. MIMAT0019739) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4665-5p," "hsa-mir-4665" (miRBase Accession No. MI0017295, SEQ ID NO: 201) is known to have a hairpin-like structure.

[0148] As used herein, the term “hsa-miR-3622a-5p gene” or “hsa-miR-3622a-5p” includes the hsa-miR-3622a-5p gene described in SEQ ID NO: 52 (miRBase Accession No. MIMAT0018003) and homologs or orthologs in other biological species. The hsa-miR-3622a-5p gene can be obtained by the method described in Witten D et al., 2010, BMC Biol, Vol. 8, p58. Furthermore, as a precursor of “hsa-miR-3622a-5p,” “hsa-mir-3622a” (miRBase Accession No. MI0016013, SEQ ID NO: 202) is known to have a hairpin-like structure.

[0149] As used herein, the term "hsa-miR-6850-5p gene" or "hsa-miR-6850-5p" includes the hsa-miR-6850-5p gene described in SEQ ID NO: 53 (miRBase Accession No. MIMAT0027600) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6850-5p", "hsa-mir-6850" (miRBase Accession No. MI0022696, SEQ ID NO: 203) is known to have a hairpin-like structure.

[0150] As used herein, the term "hsa-miR-6821-5p gene" or "hsa-miR-6821-5p" includes the hsa-miR-6821-5p gene described in SEQ ID NO: 54 (miRBase Accession No. MIMAT0027542) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6821-5p", "hsa-mir-6821" (miRBase Accession No. MI0022666, SEQ ID NO: 204) is known to have a hairpin-like structure.

[0151] As used herein, the term "hsa-miR-5100 gene" or "hsa-miR-5100" includes the hsa-miR-5100 gene described in SEQ ID NO: 55 (miRBase Accession No. MIMAT0022259) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-5100," "hsa-mir-5100" (miRBase Accession No. MI0019116, SEQ ID NO: 205) is known to have a hairpin-like structure.

[0152] As used herein, the term “hsa-miR-6872-3p gene” or “hsa-miR-6872-3p” includes the hsa-miR-6872-3p gene described in SEQ ID NO: 56 (miRBase Accession No. MIMAT0027645) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6872-3p,” “hsa-mir-6872” (miRBase Accession No. MI0022719, SEQ ID NO: 206) is known to have a hairpin-like structure.

[0153] As used herein, the term "hsa-miR-4433-3p gene" or "hsa-miR-4433-3p" includes the hsa-miR-4433-3p gene described in SEQ ID NO: 57 (miRBase Accession No. MIMAT0018949) and homologs or orthologs in other species. The hsa-miR-4433-3p gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4433-3p", "hsa-mir-4433" (miRBase Accession No. MI0016773, SEQ ID NO: 207) is known to have a hairpin-like structure.

[0154] As used herein, the term "hsa-miR-1227-5p gene" or "hsa-miR-1227-5p" includes the hsa-miR-1227-5p gene described in SEQ ID NO: 58 (miRBase Accession No. MIMAT0022941) and homologs or orthologs in other species. The hsa-miR-1227-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor of "hsa-miR-1227-5p", "hsa-mir-1227" (miRBase Accession No. MI0006316, SEQ ID NO: 208) is known to have a hairpin-like structure.

[0155] As used herein, the term “hsa-miR-3188 gene” or “hsa-miR-3188” includes the hsa-miR-3188 gene described in SEQ ID NO: 59 (miRBase Accession No. MIMAT0015070) and homologs or orthologs in other species. The hsa-miR-3188 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. Furthermore, as a precursor of “hsa-miR-3188,” “hsa-mir-3188” (miRBase Accession No. MI0014232, SEQ ID NO: 209) is known to have a hairpin-like structure.

[0156] As used herein, the term “hsa-miR-7704 gene” or “hsa-miR-7704” includes the hsa-miR-7704 gene described in SEQ ID NO: 60 (miRBase Accession No. MIMAT0030019) and homologs or orthologs in other biological species. The hsa-miR-7704 gene can be obtained by the method described in Swaminathan S et al., 2013, Biochem Biophys Res Commun, Vol. 434, pp. 228-234. In addition, as a precursor of “hsa-miR-7704,” “hsa-mir-7704” (miRBase Accession No. MI0025240, SEQ ID NO: 210) is known to form a hairpin-like structure.

[0157] As used herein, the term “hsa-miR-3185 gene” or “hsa-miR-3185” includes the hsa-miR-3185 gene described in SEQ ID NO: 61 (miRBase Accession No. MIMAT0015065) and homologs or orthologs in other species. The hsa-miR-3185 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. Furthermore, as a precursor of “hsa-miR-3185,” “hsa-mir-3185” (miRBase Accession No. MI0014227, SEQ ID NO: 211) is known to have a hairpin-like structure.

[0158] As used herein, the term “hsa-miR-1908-3p gene” or “hsa-miR-1908-3p” includes the hsa-miR-1908-3p gene described in SEQ ID NO: 62 (miRBase Accession No. MIMAT0026916) and homologs or orthologs in other biological species. 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. In addition, as a precursor of “hsa-miR-1908-3p”, “hsa-mir-1908” (miRBase Accession No. MI0008329, SEQ ID NO: 212) is known to have a hairpin-like structure.

[0159] As used herein, the term “hsa-miR-6781-5p gene” or “hsa-miR-6781-5p” includes the hsa-miR-6781-5p gene described in SEQ ID NO: 63 (miRBase Accession No. MIMAT0027462) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-6781-5p,” “hsa-mir-6781” (miRBase Accession No. MI0022626, SEQ ID NO: 213) is known to have a hairpin-like structure.

[0160] As used herein, the term "hsa-miR-6805-5p gene" or "hsa-miR-6805-5p" includes the hsa-miR-6805-5p gene described in SEQ ID NO: 64 (miRBase Accession No. MIMAT0027510) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6805-5p", "hsa-mir-6805" (miRBase Accession No. MI0022650, SEQ ID NO: 214) is known to have a hairpin-like structure.

[0161] As used herein, the term “hsa-miR-8089 gene” or “hsa-miR-8089” includes the hsa-miR-8089 gene described in SEQ ID NO: 65 (miRBase Accession No. MIMAT0031016) and homologs or orthologs in other species. The hsa-miR-8089 gene can be obtained by the method described in Wang HJ et al., 2013, Shock, Vol. 39, pp. 480-487. Furthermore, as a precursor of “hsa-miR-8089,” “hsa-mir-8089” (miRBase Accession No. MI0025925, SEQ ID NO: 215) is known to have a hairpin-like structure.

[0162] As used herein, the term "hsa-miR-665 gene" or "hsa-miR-665" includes the hsa-miR-665 gene described in SEQ ID NO: 66 (miRBase Accession No. MIMAT0004952) and homologs or orthologs in other species. 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. In addition, as a precursor of "hsa-miR-665", "hsa-mir-665" (miRBase Accession No. MI0005563, SEQ ID NO: 216) is known to have a hairpin-like structure.

[0163] As used herein, the term "hsa-miR-4486 gene" or "hsa-miR-4486" includes the hsa-miR-4486 gene described in SEQ ID NO: 67 (miRBase Accession No. MIMAT0019020) and homologs or orthologs in other species. 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 No. MI0016847, SEQ ID NO: 217) is known to have a hairpin-like structure.

[0164] As used herein, the term "hsa-miR-6722-3p gene" or "hsa-miR-6722-3p" includes the hsa-miR-6722-3p gene described in SEQ ID NO: 68 (miRBase Accession No. MIMAT0025854) and homologs or orthologs in other species. 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 of "hsa-miR-6722-3p", "hsa-mir-6722" (miRBase Accession No. MI0022557, SEQ ID NO: 218) is known to have a hairpin-like structure.

[0165] As used herein, the term “hsa-miR-1260a gene” or “hsa-miR-1260a” includes the hsa-miR-1260a gene described in SEQ ID NO: 69 (miRBase Accession No. MIMAT0005911) and homologs or orthologs in other biological species. 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 No. MI0006394, SEQ ID NO: 219) is known to have a hairpin-like structure.

[0166] As used herein, the term "hsa-miR-4707-5p gene" or "hsa-miR-4707-5p" includes the hsa-miR-4707-5p gene described in SEQ ID NO: 70 (miRBase Accession No. MIMAT0019807) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4707-5p", "hsa-mir-4707" (miRBase Accession No. MI0017340, SEQ ID NO: 220) is known to have a hairpin-like structure.

[0167] As used herein, the term "hsa-miR-6741-5p gene" or "hsa-miR-6741-5p" includes the hsa-miR-6741-5p gene described in SEQ ID NO: 71 (miRBase Accession No. MIMAT0027383) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6741-5p", "hsa-mir-6741" (miRBase Accession No. MI0022586, SEQ ID NO: 221) is known to have a hairpin-like structure.

[0168] As used herein, the term “hsa-miR-1260b gene” or “hsa-miR-1260b” includes the hsa-miR-1260b gene described in SEQ ID NO: 72 (miRBase Accession No. MIMAT0015041) and homologs or orthologs in other species. 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 No. MI0014197, SEQ ID NO: 222) is known to have a hairpin-like structure.

[0169] As used herein, the term “hsa-miR-1246 gene” or “hsa-miR-1246” includes the hsa-miR-1246 gene described in SEQ ID NO: 73 (miRBase Accession No. MIMAT0005898) and homologs or orthologs in other biological species. 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 No. MI0006381, SEQ ID NO: 223) is known to have a hairpin-like structure.

[0170] As used herein, the term "hsa-miR-6845-5p gene" or "hsa-miR-6845-5p" includes the hsa-miR-6845-5p gene described in SEQ ID NO: 74 (miRBase Accession No. MIMAT0027590) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6845-5p," "hsa-mir-6845" (miRBase Accession No. MI0022691, SEQ ID NO: 224) is known to have a hairpin-like structure.

[0171] As used herein, the term "hsa-miR-4638-5p gene" or "hsa-miR-4638-5p" includes the hsa-miR-4638-5p gene described in SEQ ID NO: 75 (miRBase Accession No. MIMAT0019695) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4638-5p", "hsa-mir-4638" (miRBase Accession No. MI0017265, SEQ ID NO: 225) is known to have a hairpin-like structure.

[0172] As used herein, the term “hsa-miR-6085 gene” or “hsa-miR-6085” includes the hsa-miR-6085 gene described in SEQ ID NO: 76 (miRBase Accession No. MIMAT0023710) and homologs or orthologs in other species. The hsa-miR-6085 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor of “hsa-miR-6085,” “hsa-mir-6085” (miRBase Accession No. MI0020362, SEQ ID NO: 226) is known to have a hairpin-like structure.

[0173] As used herein, the term "hsa-miR-1228-3p gene" or "hsa-miR-1228-3p" includes the hsa-miR-1228-3p gene described in SEQ ID NO: 77 (miRBase Accession No. MIMAT0005583) and homologs or orthologs in other species. The hsa-miR-1228-3p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as a precursor of "hsa-miR-1228-3p", "hsa-mir-1228" (miRBase Accession No. MI0006318, SEQ ID NO: 227) is known to have a hairpin-like structure.

[0174] As used herein, the term "hsa-miR-4534 gene" or "hsa-miR-4534" includes the hsa-miR-4534 gene described in SEQ ID NO: 78 (miRBase Accession No. MIMAT0019073) and homologs or orthologs in other species. The hsa-miR-4534 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4534," "hsa-mir-4534" (miRBase Accession No. MI0016901, SEQ ID NO: 228) is known to have a hairpin-like structure.

[0175] As used herein, the term "hsa-miR-5585-3p gene" or "hsa-miR-5585-3p" includes the hsa-miR-5585-3p gene described in SEQ ID NO: 79 (miRBase Accession No. MIMAT0022286) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-5585-3p," "hsa-mir-5585" (miRBase Accession No. MI0019142, SEQ ID NO: 229) is known to have a hairpin-like structure.

[0176] As used herein, the term "hsa-miR-4741 gene" or "hsa-miR-4741" includes the hsa-miR-4741 gene described in SEQ ID NO: 80 (miRBase Accession No. MIMAT0019871) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4741," "hsa-mir-4741" (miRBase Accession No. MI0017379, SEQ ID NO: 230) is known to have a hairpin-like structure.

[0177] As used herein, the term “hsa-miR-4433b-3p gene” or “hsa-miR-4433b-3p” includes the hsa-miR-4433b-3p gene described in SEQ ID NO: 81 (miRBase Accession No. MIMAT0030414) and homologs or orthologs in other species. The hsa-miR-4433b-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746. Furthermore, as a precursor of “hsa-miR-4433b-3p,” “hsa-mir-4433b” (miRBase Accession No. MI0025511, SEQ ID NO: 231) is known to have a hairpin-like structure.

[0178] As used herein, the term “hsa-miR-197-5p gene” or “hsa-miR-197-5p” includes the hsa-miR-197-5p gene described in SEQ ID NO: 82 (miRBase Accession No. MIMAT0022691) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-197-5p,” “hsa-mir-197” (miRBase Accession No. MI0000239, SEQ ID NO: 232) is known to have a hairpin-like structure.

[0179] As used herein, the term “hsa-miR-718 gene” or “hsa-miR-718” includes the hsa-miR-718 gene described in SEQ ID NO: 83 (miRBase Accession No. MIMAT0012735) and homologs or orthologs in other biological species. 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 No. MI0012489, SEQ ID NO: 233) is known to have a hairpin-like structure.

[0180] As used herein, the term "hsa-miR-4513 gene" or "hsa-miR-4513" includes the hsa-miR-4513 gene described in SEQ ID NO: 84 (miRBase Accession No. MIMAT0019050) and homologs or orthologs in other species. The hsa-miR-4513 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4513," "hsa-mir-4513" (miRBase Accession No. MI0016879, SEQ ID NO: 234) is known to have a hairpin-like structure.

[0181] As used herein, the term "hsa-miR-4446-3p gene" or "hsa-miR-4446-3p" includes the hsa-miR-4446-3p gene described in SEQ ID NO: 85 (miRBase Accession No. MIMAT0018965) and homologs or orthologs in other species. The hsa-miR-4446-3p gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4446-3p", "hsa-mir-4446" (miRBase Accession No. MI0016789, SEQ ID NO: 235) is known to have a hairpin-like structure.

[0182] As used herein, the term "hsa-miR-619-5p gene" or "hsa-miR-619-5p" includes the hsa-miR-619-5p gene described in SEQ ID NO: 86 (miRBase Accession No. MIMAT0026622) and homologs or orthologs in other species. The hsa-miR-619-5p 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 of "hsa-miR-619-5p", "hsa-mir-619" (miRBase Accession No. MI0003633, SEQ ID NO: 236) is known to have a hairpin-like structure.

[0183] As used herein, the term "hsa-miR-6816-5p gene" or "hsa-miR-6816-5p" includes the hsa-miR-6816-5p gene described in SEQ ID NO: 87 (miRBase Accession No. MIMAT0027532) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6816-5p", "hsa-mir-6816" (miRBase Accession No. MI0022661, SEQ ID NO: 237) is known to have a hairpin-like structure.

[0184] As used herein, the term “hsa-miR-6778-5p gene” or “hsa-miR-6778-5p” includes the hsa-miR-6778-5p gene described in SEQ ID NO: 88 (miRBase Accession No. MIMAT0027456) and homologs or orthologs in other species. 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 of “hsa-miR-6778-5p,” “hsa-mir-6778” (miRBase Accession No. MI0022623, SEQ ID NO: 238) is known to have a hairpin-like structure.

[0185] As used herein, the term “hsa-miR-24-3p gene” or “hsa-miR-24-3p” includes the hsa-miR-24-3p gene described in SEQ ID NO: 89 (miRBase Accession No. MIMAT0000080) and homologs or orthologs in other biological species. 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 of “hsa-miR-24-3p”, “hsa-mir-24-1” and “hsa-mir-24-2” (miRBase Accession Nos. MI0000080, MI0000081, SEQ ID NOs: 239, 240) are known as hairpin-like structures.

[0186] As used herein, the term "hsa-miR-1915-3p gene" or "hsa-miR-1915-3p" includes the hsa-miR-1915-3p gene described in SEQ ID NO: 90 (miRBase Accession No. MIMAT0007892) and homologs or orthologs in other biological species. 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 of "hsa-miR-1915-3p", "hsa-mir-1915" (miRBase Accession No. MI0008336, SEQ ID NO: 241) is known to have a hairpin-like structure.

[0187] As used herein, the term "hsa-miR-4665-3p gene" or "hsa-miR-4665-3p" includes the hsa-miR-4665-3p gene described in SEQ ID NO: 91 (miRBase Accession No. MIMAT0019740) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4665-3p", "hsa-mir-4665" (miRBase Accession No. MI0017295, SEQ ID NO: 201) is known to have a hairpin-like structure.

[0188] As used herein, the term "hsa-miR-4449 gene" or "hsa-miR-4449" includes the hsa-miR-4449 gene described in SEQ ID NO: 92 (miRBase Accession No. MIMAT0018968) and homologs or orthologs in other species. The hsa-miR-4449 gene can be obtained by the method described in Jima DD et al., 2010, Blood, Vol. 116, e118-e127. Furthermore, as a precursor of "hsa-miR-4449", "hsa-mir-4449" (miRBase Accession No. MI0016792, SEQ ID NO: 242) is known to have a hairpin-like structure.

[0189] As used herein, the term “hsa-miR-6889-5p gene” or “hsa-miR-6889-5p” includes the hsa-miR-6889-5p gene described in SEQ ID NO: 93 (miRBase Accession No. MIMAT0027678) and homologs or orthologs in other species. 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 of “hsa-miR-6889-5p,” “hsa-mir-6889” (miRBase Accession No. MI0022736, SEQ ID NO: 243) is known to have a hairpin-like structure.

[0190] As used herein, the term "hsa-miR-486-3p gene" or "hsa-miR-486-3p" includes the hsa-miR-486-3p gene described in SEQ ID NO: 94 (miRBase Accession No. MIMAT0004762) and homologs or orthologs in other species. 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. Furthermore, as precursors of "hsa-miR-486-3p", "hsa-mir-486, hsa-mir-486-2" (miRBase Accession Nos. MI0002470, MI0023622, SEQ ID NOs. 244, 245) are known.

[0191] As used herein, the term "hsa-miR-7113-3p gene" or "hsa-miR-7113-3p" includes the hsa-miR-7113-3p gene described in SEQ ID NO: 95 (miRBase Accession No. MIMAT0028124) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-7113-3p", "hsa-mir-7113" (miRBase Accession No. MI0022964, SEQ ID NO: 246) is known to have a hairpin-like structure.

[0192] As used herein, the term "hsa-miR-642a-3p gene" or "hsa-miR-642a-3p" includes the hsa-miR-642a-3p gene described in SEQ ID NO: 96 (miRBase Accession No. MIMAT0020924) and homologs or orthologs in other species. 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 of "hsa-miR-642a-3p", "hsa-mir-642a" (miRBase Accession No. MI0003657, SEQ ID NO: 247) is known to have a hairpin-like structure.

[0193] As used herein, the term "hsa-miR-7847-3p gene" or "hsa-miR-7847-3p" includes the hsa-miR-7847-3p gene described in SEQ ID NO: 97 (miRBase Accession No. MIMAT0030422) and homologs or orthologs in other species. The hsa-miR-7847-3p gene can be obtained by the method described in Ple H et al., 2012, PLoS One, Vol. 7, e50746. Furthermore, as a precursor of "hsa-miR-7847-3p", "hsa-mir-7847" (miRBase Accession No. MI0025517, SEQ ID NO: 248) is known to have a hairpin-like structure.

[0194] As used herein, the term "hsa-miR-6768-5p gene" or "hsa-miR-6768-5p" includes the hsa-miR-6768-5p gene described in SEQ ID NO: 98 (miRBase Accession No. MIMAT0027436) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-6768-5p", "hsa-mir-6768" (miRBase Accession No. MI0022613, SEQ ID NO: 249) is known to have a hairpin-like structure.

[0195] As used herein, the term “hsa-miR-1290 gene” or “hsa-miR-1290” includes the hsa-miR-1290 gene described in SEQ ID NO: 99 (miRBase Accession No. MIMAT0005880) and homologs or orthologs in other biological species. 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 No. MI0006352, SEQ ID NO: 250) is known to have a hairpin-like structure.

[0196] As used herein, the term "hsa-miR-7108-5p gene" or "hsa-miR-7108-5p" includes the hsa-miR-7108-5p gene described in SEQ ID NO: 100 (miRBase Accession No. MIMAT0028113) and homologs or orthologs in other species. The hsa-miR-7108-5p 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 of "hsa-miR-7108-5p", "hsa-mir-7108" (miRBase Accession No. MI0022959, SEQ ID NO: 251) is known to have a hairpin-like structure.

[0197] As used herein, the term "hsa-miR-92b-5p gene" or "hsa-miR-92b-5p" includes the hsa-miR-92b-5p gene described in SEQ ID NO: 101 (miRBase Accession No. MIMAT0004792) and homologs or orthologs in other species. The hsa-miR-92b-5p 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 of "hsa-miR-92b-5p", "hsa-mir-92b" (miRBase Accession No. MI0003560, SEQ ID NO: 252) is known to have a hairpin-like structure.

[0198] As used herein, the term "hsa-miR-663b gene" or "hsa-miR-663b" includes the hsa-miR-663b gene described in SEQ ID NO: 102 (miRBase Accession No. MIMAT0005867) and homologs or orthologs in other species. The hsa-miR-663b gene can be obtained by the method described in Takada S et al., 2008, Leukemia, Vol. 22, pp. 1274-1278. Furthermore, as a precursor of "hsa-miR-663b," "hsa-mir-663b" (miRBase Accession No. MI0006336, SEQ ID NO: 253) is known to have a hairpin-like structure.

[0199] As used herein, the term "hsa-miR-3940-5p gene" or "hsa-miR-3940-5p" includes the hsa-miR-3940-5p gene described in SEQ ID NO: 103 (miRBase Accession No. MIMAT0019229) and homologs or orthologs in other species. The hsa-miR-3940-5p gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, Vol. 5, e10563. Furthermore, as a precursor of "hsa-miR-3940-5p," "hsa-mir-3940" (miRBase Accession No. MI0016597, SEQ ID NO: 254) is known to have a hairpin-like structure.

[0200] As used herein, the term "hsa-miR-4467 gene" or "hsa-miR-4467" includes the hsa-miR-4467 gene described in SEQ ID NO: 104 (miRBase Accession No. MIMAT0018994) and homologs or orthologs in other species. 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 No. MI0016818, SEQ ID NO: 255) is known to have a hairpin-like structure.

[0201] As used herein, the term "hsa-miR-6858-5p gene" or "hsa-miR-6858-5p" includes the hsa-miR-6858-5p gene described in SEQ ID NO: 105 (miRBase Accession No. MIMAT0027616) and homologs or orthologs in other species. The hsa-miR-6858-5p 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 of "hsa-miR-6858-5p", "hsa-mir-6858" (miRBase Accession No. MI0022704, SEQ ID NO: 256) is known to have a hairpin-like structure.

[0202] As used herein, the term "hsa-miR-4417 gene" or "hsa-miR-4417" includes the hsa-miR-4417 gene described in SEQ ID NO: 106 (miRBase Accession No. MIMAT0018929) and homologs or orthologs in other species. 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 No. MI0016753, SEQ ID NO: 257) is known to have a hairpin-like structure.

[0203] As used herein, the term "hsa-miR-3665 gene" or "hsa-miR-3665" includes the hsa-miR-3665 gene described in SEQ ID NO: 107 (miRBase Accession No. MIMAT0018087) and homologs or orthologs in other species. The hsa-miR-3665 gene can be obtained by the method described in Xie X et al., 2005, Nature, Vol. 434, pp. 338-345. Furthermore, as a precursor of "hsa-miR-3665," "hsa-mir-3665" (miRBase Accession No. MI0016066, SEQ ID NO: 258) is known to have a hairpin-like structure.

[0204] As used herein, the term "hsa-miR-4736 gene" or "hsa-miR-4736" includes the hsa-miR-4736 gene described in SEQ ID NO: 108 (miRBase Accession No. MIMAT0019862) and homologs or orthologs in other species. The hsa-miR-4736 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 of "hsa-miR-4736," "hsa-mir-4736" (miRBase Accession No. MI0017373, SEQ ID NO: 259) is known to have a hairpin-like structure.

[0205] As used herein, the term "hsa-miR-4687-3p gene" or "hsa-miR-4687-3p" includes the hsa-miR-4687-3p gene described in SEQ ID NO: 109 (miRBase Accession No. MIMAT0019775) and homologs or orthologs in other species. The hsa-miR-4687-3p 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 of "hsa-miR-4687-3p", "hsa-mir-4687" (miRBase Accession No. MI0017319, SEQ ID NO: 260) is known to have a hairpin-like structure.

[0206] As used herein, the term “hsa-miR-1908-5p gene” or “hsa-miR-1908-5p” includes the hsa-miR-1908-5p gene described in SEQ ID NO: 110 (miRBase Accession No. MIMAT0007881) and homologs or orthologs in other biological species. 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 of “hsa-miR-1908-5p,” “hsa-mir-1908” (miRBase Accession No. MI0008329, SEQ ID NO: 212) is known to have a hairpin-like structure.

[0207] As used herein, the term "hsa-miR-5195-3p gene" or "hsa-miR-5195-3p" includes the hsa-miR-5195-3p gene described in SEQ ID NO: 111 (miRBase Accession No. MIMAT0021127) and homologs or orthologs in other species. 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 of "hsa-miR-5195-3p," "hsa-mir-5195" (miRBase Accession No. MI0018174, SEQ ID NO: 261) is known to have a hairpin-like structure.

[0208] As used herein, the term “hsa-miR-4286 gene” or “hsa-miR-4286” includes the hsa-miR-4286 gene described in SEQ ID NO: 112 (miRBase Accession No. MIMAT0016916) and homologs or orthologs in other species. 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 No. MI0015894, SEQ ID NO: 262) is known to have a hairpin-like structure.

[0209] As used herein, the term "hsa-miR-3679-3p gene" or "hsa-miR-3679-3p" includes the hsa-miR-3679-3p gene described in SEQ ID NO: 113 (miRBase Accession No. MIMAT0018105) and homologs or orthologs in other species. The hsa-miR-3679-3p gene can be obtained by the method described in Creighton CJ et al., 2010, PLoS One, Vol. 5, e9637. Furthermore, as a precursor of "hsa-miR-3679-3p", "hsa-mir-3679" (miRBase Accession No. MI0016080, SEQ ID NO: 263) is known to have a hairpin-like structure.

[0210] As used herein, the term "hsa-miR-6791-5p gene" or "hsa-miR-6791-5p" includes the hsa-miR-6791-5p gene described in SEQ ID NO: 114 (miRBase Accession No. MIMAT0027482) and homologs or orthologs in other species. The hsa-miR-6791-5p 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 of "hsa-miR-6791-5p", "hsa-mir-6791" (miRBase Accession No. MI0022636, SEQ ID NO: 264) is known to have a hairpin-like structure.

[0211] As used herein, the term "hsa-miR-1202 gene" or "hsa-miR-1202" includes the hsa-miR-1202 gene described in SEQ ID NO: 115 (miRBase Accession No. MIMAT0005865) and homologs or orthologs in other species. 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 of "hsa-miR-1202," "hsa-mir-1202" (miRBase Accession No. MI0006334, SEQ ID NO: 265) is known to have a hairpin-like structure.

[0212] As used herein, the term "hsa-miR-3656 gene" or "hsa-miR-3656" includes the hsa-miR-3656 gene described in SEQ ID NO: 116 (miRBase Accession No. MIMAT0018076) and homologs or orthologs in other species. 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 of "hsa-miR-3656", "hsa-mir-3656" (miRBase Accession No. MI0016056, SEQ ID NO: 266) is known to have a hairpin-like structure.

[0213] As used herein, the term "hsa-miR-4746-3p gene" or "hsa-miR-4746-3p" includes the hsa-miR-4746-3p gene described in SEQ ID NO: 117 (miRBase Accession No. MIMAT0019881) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-4746-3p", "hsa-mir-4746" (miRBase Accession No. MI0017385, SEQ ID NO: 267) is known to have a hairpin-like structure.

[0214] As used herein, the term “hsa-miR-3184-5p gene” or “hsa-miR-3184-5p” includes the hsa-miR-3184-5p gene described in SEQ ID NO: 118 (miRBase Accession No. MIMAT0015064) and homologs or orthologs in other species. The hsa-miR-3184-5p gene can be obtained by the method described in Stark MS et al., 2010, PLoS One, Vol. 5, e9685. Furthermore, as a precursor of “hsa-miR-3184-5p,” “hsa-mir-3184” (miRBase Accession No. MI0014226, SEQ ID NO: 268) is known to have a hairpin-like structure.

[0215] As used herein, the term "hsa-miR-3937 gene" or "hsa-miR-3937" includes the hsa-miR-3937 gene described in SEQ ID NO: 119 (miRBase Accession No. MIMAT0018352) and homologs or orthologs in other species. The hsa-miR-3937 gene can be obtained by the method described in Liao JY et al., 2010, PLoS One, Vol. 5, e10563. Furthermore, as a precursor of "hsa-miR-3937," "hsa-mir-3937" (miRBase Accession No. MI0016593, SEQ ID NO: 269) is known to have a hairpin-like structure.

[0216] As used herein, the term "hsa-miR-6515-3p gene" or "hsa-miR-6515-3p" includes the hsa-miR-6515-3p gene described in SEQ ID NO: 120 (miRBase Accession No. MIMAT0025487) and homologs or orthologs in other species. The hsa-miR-6515-3p gene can be obtained by the method described in Joyce CE et al., 2011, Hum Mol Genet, Vol. 20, pp. 4025-4040. Furthermore, as a precursor of "hsa-miR-6515-3p", "hsa-mir-6515" (miRBase Accession No. MI0022227, SEQ ID NO: 270) is known to have a hairpin-like structure.

[0217] As used herein, the term “hsa-miR-6132 gene” or “hsa-miR-6132” includes the hsa-miR-6132 gene described in SEQ ID NO: 121 (miRBase Accession No. MIMAT0024616) and homologs or orthologs in other species. The hsa-miR-6132 gene can be obtained by the method described in Dannemann M et al., 2012, Genome Biol Evol, 4, pp. 552-564. Furthermore, as a precursor of “hsa-miR-6132,” “hsa-mir-6132” (miRBase Accession No. MI0021277, SEQ ID NO: 271) is known to have a hairpin-like structure.

[0218] As used herein, the term "hsa-miR-187-5p gene" or "hsa-miR-187-5p" includes the hsa-miR-187-5p gene described in SEQ ID NO: 122 (miRBase Accession No. MIMAT0004561) and homologs or orthologs in other species. The hsa-miR-187-5p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. Furthermore, as a precursor of "hsa-miR-187-5p", "hsa-mir-187" (miRBase Accession No. MI0000274, SEQ ID NO: 272) is known to have a hairpin-like structure.

[0219] As used herein, the term "hsa-miR-7111-5p gene" or "hsa-miR-7111-5p" includes the hsa-miR-7111-5p gene described in SEQ ID NO: 123 (miRBase Accession No. MIMAT0028119) and homologs or orthologs in other species. The hsa-miR-7111-5p 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 of "hsa-miR-7111-5p", "hsa-mir-7111" (miRBase Accession No. MI0022962, SEQ ID NO: 273) is known to have a hairpin-like structure.

[0220] As used herein, the term “hsa-miR-5787 gene” or “hsa-miR-5787” includes the hsa-miR-5787 gene described in SEQ ID NO: 124 (miRBase Accession No. MIMAT0023252) and homologs or orthologs in other species. 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 of “hsa-miR-5787,” “hsa-mir-5787” (miRBase Accession No. MI0019797, SEQ ID NO: 274) is known to have a hairpin-like structure.

[0221] As used herein, the term "hsa-miR-6779-5p gene" or "hsa-miR-6779-5p" includes the hsa-miR-6779-5p gene described in SEQ ID NO: 125 (miRBase Accession No. MIMAT0027458) and homologs or orthologs in other species. The hsa-miR-6779-5p 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 of "hsa-miR-6779-5p", "hsa-mir-6779" (miRBase Accession No. MI0022624, SEQ ID NO: 275) is known to have a hairpin-like structure.

[0222] As used herein, the term "hsa-miR-6808-5p gene" or "hsa-miR-6808-5p" includes the hsa-miR-6808-5p gene described in SEQ ID NO: 126 (miRBase Accession No. MIMAT0027516) and homologs or orthologs in other species. The hsa-miR-6808-5p 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 of "hsa-miR-6808-5p", "hsa-mir-6808" (miRBase Accession No. MI0022653, SEQ ID NO: 276) is known to have a hairpin-like structure.

[0223] As used herein, the term "hsa-miR-6774-5p gene" or "hsa-miR-6774-5p" includes the hsa-miR-6774-5p gene described in SEQ ID NO: 127 (miRBase Accession No. MIMAT0027448) and homologs or orthologs in other species. The hsa-miR-6774-5p 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 of "hsa-miR-6774-5p", "hsa-mir-6774" (miRBase Accession No. MI0022619, SEQ ID NO: 277) is known to have a hairpin-like structure.

[0224] As used herein, the term “hsa-miR-4656 gene” or “hsa-miR-4656” includes the hsa-miR-4656 gene described in SEQ ID NO: 128 (miRBase Accession No. MIMAT0019723) and homologs or orthologs in other species. The hsa-miR-4656 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 of “hsa-miR-4656,” “hsa-mir-4656” (miRBase Accession No. MI0017284, SEQ ID NO: 278) is known to have a hairpin-like structure.

[0225] As used herein, the term "hsa-miR-6806-5p gene" or "hsa-miR-6806-5p" includes the hsa-miR-6806-5p gene described in SEQ ID NO: 129 (miRBase Accession No. MIMAT0027512) and homologs or orthologs in other species. The hsa-miR-6806-5p 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 of "hsa-miR-6806-5p", "hsa-mir-6806" (miRBase Accession No. MI0022651, SEQ ID NO: 279) is known to have a hairpin-like structure.

[0226] As used herein, the term "hsa-miR-1233-5p gene" or "hsa-miR-1233-5p" includes the hsa-miR-1233-5p gene described in SEQ ID NO: 130 (miRBase Accession No. MIMAT0022943) and homologs or orthologs in other biological species. The hsa-miR-1233-5p gene can be obtained by the method described in Berezikov E et al., 2007, Mol Cell, Vol. 28, pp. 328-336. Furthermore, as precursors of "hsa-miR-1233-5p", "hsa-mir-1233-1" and "hsa-mir-1233-2" (miRBase Accession Nos. MI0006323 and MI0015973, SEQ ID NOs: 280 and 281) are known.

[0227] As used herein, the term "hsa-miR-328-5p gene" or "hsa-miR-328-5p" includes the hsa-miR-328-5p gene described in SEQ ID NO: 131 (miRBase Accession No. MIMAT0026486) and homologs or orthologs in other species. 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 of "hsa-miR-328-5p", "hsa-mir-328" (miRBase Accession No. MI0000804, SEQ ID NO: 282) is known to have a hairpin-like structure.

[0228] As used herein, the term "hsa-miR-4674 gene" or "hsa-miR-4674" includes the hsa-miR-4674 gene described in SEQ ID NO: 132 (miRBase Accession No. MIMAT0019756) and homologs or orthologs in other species. The hsa-miR-4674 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-4674", "hsa-mir-4674" (miRBase Accession No. MI0017305, SEQ ID NO: 283) is known to have a hairpin-like structure.

[0229] As used herein, the term “hsa-miR-2110 gene” or “hsa-miR-2110” includes the hsa-miR-2110 gene described in SEQ ID NO: 133 (miRBase Accession No. MIMAT0010133) as well as homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-2110,” “hsa-mir-2110” (miRBase Accession No. MI0010629, SEQ ID NO: 284) is known to have a hairpin-like structure.

[0230] As used herein, the term “hsa-miR-6076 gene” or “hsa-miR-6076” includes the hsa-miR-6076 gene described in SEQ ID NO: 134 (miRBase Accession No. MIMAT0023701) and homologs or orthologs in other species. The hsa-miR-6076 gene can be obtained by the method described in Voellenkle C et al., 2012, RNA, Vol. 18, pp. 472-484. Furthermore, as a precursor of “hsa-miR-6076,” “hsa-mir-6076” (miRBase Accession No. MI0020353, SEQ ID NO: 285) is known to have a hairpin-like structure.

[0231] As used herein, the term "hsa-miR-3619-3p gene" or "hsa-miR-3619-3p" includes the hsa-miR-3619-3p gene described in SEQ ID NO: 135 (miRBase Accession No. MIMAT0019219) and homologs or orthologs in other biological species. The hsa-miR-3619-3p gene can be obtained by the method described in Witten D et al., 2010, BMC Biol, Vol. 8, p58. Furthermore, as a precursor of "hsa-miR-3619-3p", "hsa-mir-3619" (miRBase Accession No. MI0016009, SEQ ID NO: 286) is known to have a hairpin-like structure.

[0232] As used herein, the term "hsa-miR-92a-2-5p gene" or "hsa-miR-92a-2-5p" includes the hsa-miR-92a-2-5p gene described in SEQ ID NO: 136 (miRBase Accession No. MIMAT0004508) and homologs or orthologs in other biological species. 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. In addition, as a precursor of "hsa-miR-92a-2-5p", "hsa-mir-92a-2" (miRBase Accession No. MI0000094, SEQ ID NO: 287) is known to have a hairpin-like structure.

[0233] As used herein, the term "hsa-miR-128-1-5p gene" or "hsa-miR-128-1-5p" includes the hsa-miR-128-1-5p gene described in SEQ ID NO: 137 (miRBase Accession No. MIMAT0026477) and homologs or orthologs in other biological species. 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. Furthermore, as a precursor of "hsa-miR-128-1-5p", "hsa-mir-128-1" (miRBase Accession No. MI0000447, SEQ ID NO: 288) is known to have a hairpin-like structure.

[0234] As used herein, the term "hsa-miR-638 gene" or "hsa-miR-638" includes the hsa-miR-638 gene described in SEQ ID NO: 138 (miRBase Accession No. MIMAT0003308) and homologs or orthologs in other species. 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, p3687-3692. In addition, as a precursor of "hsa-miR-638", "hsa-mir-638" (miRBase Accession No. MI0003653, SEQ ID NO: 289) is known to have a hairpin-like structure.

[0235] As used herein, the term "hsa-miR-2861 gene" or "hsa-miR-2861" includes the hsa-miR-2861 gene described in SEQ ID NO: 139 (miRBase Accession No. MIMAT0013802) and homologs or orthologs in other species. 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 of "hsa-miR-2861," "hsa-mir-2861" (miRBase Accession No. MI0013006, SEQ ID NO: 290) is known to have a hairpin-like structure.

[0236] As used herein, the term "hsa-miR-371a-5p gene" or "hsa-miR-371a-5p" includes the hsa-miR-371a-5p gene described in SEQ ID NO: 140 (miRBase Accession No. MIMAT0004687) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-371a-5p", "hsa-mir-371a" (miRBase Accession No. MI0000779, SEQ ID NO: 291) is known to have a hairpin-like structure.

[0237] As used herein, the term "hsa-miR-211-3p gene" or "hsa-miR-211-3p" includes the hsa-miR-211-3p gene described in SEQ ID NO: 141 (miRBase Accession No. MIMAT0022694) and homologs or orthologs in other species. The hsa-miR-211-3p gene can be obtained by the method described in Lim LP et al., 2003, Science, Vol. 299, p1540. Furthermore, as a precursor of "hsa-miR-211-3p", "hsa-mir-211" (miRBase Accession No. MI0000287, SEQ ID NO: 292) is known, which has a hairpin-like structure.

[0238] As used herein, the term “hsa-miR-1273g-3p gene” or “hsa-miR-1273g-3p” includes the hsa-miR-1273g-3p gene described in SEQ ID NO: 142 (miRBase Accession No. MIMAT0022742) and homologs or orthologs in other biological species. 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 of “hsa-miR-1273g-3p,” “hsa-mir-1273g” (miRBase Accession No. MI0018003, SEQ ID NO: 293) is known to have a hairpin-like structure.

[0239] As used herein, the term "hsa-miR-1203 gene" or "hsa-miR-1203" includes the hsa-miR-1203 gene described in SEQ ID NO: 143 (miRBase Accession No. MIMAT0005866) and homologs or orthologs in other species. The hsa-miR-1203 gene can be obtained by the method described in Marton S et al., 2008, Leukemia, Vol. 22, pp. 330-338. Furthermore, as a precursor of "hsa-miR-1203," "hsa-mir-1203" (miRBase Accession No. MI0006335, SEQ ID NO: 294) is known to have a hairpin-like structure.

[0240] As used herein, the term "hsa-miR-122-5p gene" or "hsa-miR-122-5p" includes the hsa-miR-122-5p gene described in SEQ ID NO: 144 (miRBase Accession No. MIMAT0000421) and homologs or orthologs in other species. The hsa-miR-122-5p gene can be obtained by the method described in Lagos-Quintana M et al., 2002, Curr Biol, Vol. 12, pp. 735-739. Furthermore, as a precursor of "hsa-miR-122-5p", "hsa-mir-122" (miRBase Accession No. MI0000442, SEQ ID NO: 295) is known to have a hairpin-like structure.

[0241] As used herein, the term “hsa-miR-4258 gene” or “hsa-miR-4258” includes the hsa-miR-4258 gene described in SEQ ID NO: 145 (miRBase Accession No. MIMAT0016879) and homologs or orthologs in other species. 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 No. MI0015857, SEQ ID NO: 296) is known to have a hairpin-like structure.

[0242] As used herein, the term "hsa-miR-4484 gene" or "hsa-miR-4484" includes the hsa-miR-4484 gene described in SEQ ID NO: 146 (miRBase Accession No. MIMAT0019018) and homologs or orthologs in other species. 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 No. MI0016845, SEQ ID NO: 297) is known to have a hairpin-like structure.

[0243] As used herein, the term “hsa-miR-4648 gene” or “hsa-miR-4648” includes the hsa-miR-4648 gene described in SEQ ID NO: 147 (miRBase Accession No. MIMAT0019710) and homologs or orthologs in other species. 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. Furthermore, as a precursor of “hsa-miR-4648,” “hsa-mir-4648” (miRBase Accession No. MI0017275, SEQ ID NO: 298) is known to have a hairpin-like structure.

[0244] As used herein, the term “hsa-miR-6780b-5p gene” or “hsa-miR-6780b-5p” includes the hsa-miR-6780b-5p gene described in SEQ ID NO: 148 (miRBase Accession No. MIMAT0027572) and homologs or orthologs in other species. The hsa-miR-6780b-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 of “hsa-miR-6780b-5p,” “hsa-mir-6780b” (miRBase Accession No. MI0022681, SEQ ID NO: 299) is known to have a hairpin-like structure.

[0245] As used herein, the term "hsa-miR-4516 gene" or "hsa-miR-4516" includes the hsa-miR-4516 gene described in SEQ ID NO: 466 (miRBase Accession No. MIMAT0019053) and homologs or orthologs in other species. 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 of "hsa-miR-4516", "hsa-mir-4516" (miRBase Accession No. MI0016882, SEQ ID NO: 479) is known to have a hairpin-like structure.

[0246] As used herein, the term "hsa-miR-4649-5p gene" or "hsa-miR-4649-5p" includes the hsa-miR-4649-5p gene described in SEQ ID NO: 467 (miRBase Accession No. MIMAT0019711) and homologs or orthologs in other species. 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 of "hsa-miR-4649-5p", "hsa-mir-4649" (miRBase Accession No. MI0017276, SEQ ID NO: 480) is known to have a hairpin-like structure.

[0247] As used herein, the term "hsa-miR-760 gene" or "hsa-miR-760" includes the hsa-miR-760 gene described in SEQ ID NO: 468 (miRBase Accession No. MIMAT0004957) and homologs or orthologs in other species. 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. In addition, as a precursor of "hsa-miR-760", "hsa-mir-760" (miRBase Accession No. MI0005567, SEQ ID NO: 481) is known to have a hairpin-like structure.

[0248] As used herein, the term “hsa-miR-3162-5p gene” or “hsa-miR-3162-5p” includes the hsa-miR-3162-5p gene described in SEQ ID NO: 469 (miRBase Accession No. MIMAT0015036) as well as homologs or orthologs in other species. The hsa-miR-3162-5p gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. Furthermore, as a precursor of “hsa-miR-3162-5p,” “hsa-mir-3162” (miRBase Accession No. MI0014192, SEQ ID NO: 482) is known to have a hairpin-like structure.

[0249] As used herein, the term “hsa-miR-3178 gene” or “hsa-miR-3178” includes the hsa-miR-3178 gene described in SEQ ID NO: 470 (miRBase Accession No. MIMAT0015055) and homologs or orthologs in other species. The hsa-miR-3178 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. Furthermore, as a precursor of “hsa-miR-3178,” “hsa-mir-3178” (miRBase Accession No. MI0014212, SEQ ID NO: 483) is known to have a hairpin-like structure.

[0250] As used herein, the term "hsa-miR-940 gene" or "hsa-miR-940" includes the hsa-miR-940 gene described in SEQ ID NO: 471 (miRBase Accession No. MIMAT0004983) and homologs or orthologs in other species. The hsa-miR-940 gene can be obtained by the method described in Lui WO et al., 2007, Cancer Res., Vol. 67, pp. 6031-6043. Furthermore, as a precursor of "hsa-miR-940," "hsa-mir-940" (miRBase Accession No. MI0005762, SEQ ID NO: 484) is known to have a hairpin-like structure.

[0251] As used herein, the term “hsa-miR-4271 gene” or “hsa-miR-4271” includes the hsa-miR-4271 gene described in SEQ ID NO: 472 (miRBase Accession No. MIMAT0016901) and homologs or orthologs in other species. 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 No. MI0015879, SEQ ID NO: 485) is known to have a hairpin-like structure.

[0252] As used herein, the term "hsa-miR-6769b-5p gene" or "hsa-miR-6769b-5p" includes the hsa-miR-6769b-5p gene described in SEQ ID NO: 473 (miRBase Accession No. MIMAT0027620) and homologs or orthologs in other species. 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 of "hsa-miR-6769b-5p", "hsa-mir-6769b" (miRBase Accession No. MI0022706, SEQ ID NO: 486) is known to have a hairpin-like structure.

[0253] As used herein, the term "hsa-miR-4508 gene" or "hsa-miR-4508" includes the hsa-miR-4508 gene described in SEQ ID NO: 474 (miRBase Accession No. MIMAT0019045) and homologs or orthologs in other species. 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 of "hsa-miR-4508," "hsa-mir-4508" (miRBase Accession No. MI0016872, SEQ ID NO: 487) is known to have a hairpin-like structure.

[0254] As used herein, the term "hsa-miR-6826-5p gene" or "hsa-miR-6826-5p" includes the hsa-miR-6826-5p gene described in SEQ ID NO: 475 (miRBase Accession No. MIMAT0027552) and homologs or orthologs in other species. 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 of "hsa-miR-6826-5p", "hsa-mir-6826" (miRBase Accession No. MI0022671, SEQ ID NO: 488) is known to have a hairpin-like structure.

[0255] As used herein, the term "hsa-miR-6757-5p gene" or "hsa-miR-6757-5p" includes the hsa-miR-6757-5p gene described in SEQ ID NO: 476 (miRBase Accession No. MIMAT0027414) and homologs or orthologs in other species. 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 of "hsa-miR-6757-5p", "hsa-mir-6757" (miRBase Accession No. MI0022602, SEQ ID NO: 489) is known to have a hairpin-like structure.

[0256] As used herein, the term "hsa-miR-3131 gene" or "hsa-miR-3131" includes the hsa-miR-3131 gene described in SEQ ID NO: 477 (miRBase Accession No. MIMAT0014996) and homologs or orthologs in other species. The hsa-miR-3131 gene can be obtained by the method described in Stark MS et al., 2010, PLoS One., Vol. 5, e9685. Furthermore, as a precursor of "hsa-miR-3131," "hsa-mir-3131" (miRBase Accession No. MI0014151, SEQ ID NO: 490) is known to have a hairpin-like structure.

[0257] As used herein, the term "hsa-miR-1343-3p gene" or "hsa-miR-1343-3p" includes the hsa-miR-1343-3p gene described in SEQ ID NO: 478 (miRBase Accession No. MIMAT0019776) and homologs or orthologs in other species. 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. Furthermore, as a precursor of "hsa-miR-1343-3p", "hsa-mir-1343" (miRBase Accession No. MI0017320, SEQ ID NO: 491) is known to have a hairpin-like structure.

[0258] In addition, when a mature miRNA is cut out from an RNA precursor that forms a hairpin-like structure as a mature miRNA, sometimes one to several bases before and after the sequence are cut out short or long, and base substitution occurs to produce a mutant, which is called isomiR (Morin RD. et al., 2008, Genome Research, Vol. 18, p. 610-621). In miRBase version 20, in addition to the base sequences shown in any one of sequence numbers 1 to 148, 466 to 478, a plurality of mutants and fragments of the base sequences shown in any one of sequence numbers 300 to 465 and 492 to 509, which are called isomiR, are also shown. These mutants can also be obtained as miRNAs having the base sequences shown in any one of sequence numbers 1 to 148, 466 to 478.

[0259] That is, the 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, 84, 85, 86, 89, 90, 92, 94, 96, 99, 101, 102, 103, 104, 106, 107, 109, 110 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 a polynucleotide consisting of a base sequence in which u is t in the base sequence, as the longest variant registered in, for example, miRBase version 20, Serial numbers 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 , 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] In addition, 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, 110 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 a polynucleotide consisting of a base sequence in which u is t in the base sequence, as the shortest variant registered in miRBase version 20, for example, Serial numbers 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, 392 499, 495, 497, 499, 501, 503, 505, 507, and 509.In addition to these mutants and fragments, 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, 84 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. Furthermore, examples of polynucleotides comprising the base sequence set forth in any one of SEQ ID NOs: 1 to 148 and 466 to 478 include the polynucleotides set forth in any one of SEQ ID NOs: 149 to 299 and 479 to 491 as precursors, respectively.

[0261] Table 1 lists the names and miRBase accession numbers (accession numbers) of the genes represented by SEQ ID NOs: 1 to 509.

[0262] In this specification, "capable of specifically binding" means that the nucleic acid probe or primer used in the present invention binds to a specific target nucleic acid and is substantially unable to 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 are the bases of priority of this application.

[0280] Effects of the Invention

[0281] The present invention makes it possible to easily and accurately detect biliary tract cancer. For example, using the measured values of several miRNAs in the patient's blood, serum, and / or plasma, which can be collected minimally invasively, as indicators, it is possible to easily detect whether the patient has biliary tract cancer. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0283] Figure 2 Middle, left: A graph showing the measured values of hsa-miR-125a-3p (sequence number 1) of healthy subjects (100 people) and biliary tract cancer patients (67 people) selected as the learning sample group as the vertical axis. The horizontal line in the figure represents the threshold value (5.69) for distinguishing the two groups optimized by Fisher's discriminant analysis. Right: A graph showing the measured values of hsa-miR-125a-3p (sequence number 1) of healthy subjects (50 people) and biliary tract cancer patients (33 people) selected as the test sample group as the vertical axis. The horizontal line in the figure represents the threshold value (5.69) for distinguishing the two groups set by the learning sample group.

[0284] Figure 3Middle, left figure: A figure showing the measured values of hsa-miR-6893-5p (sequence number 2) of healthy subjects (100 people, circles) and biliary tract cancer patients (67 people, triangles) selected as the learning sample group as the horizontal axis and the measured values of hsa-miR-4476 (sequence number 4) as the vertical axis. The line in the figure represents the discriminant function (0 = 5.16x + y + 48.11) for discriminating the two groups, which was optimized by Fisher's discriminant analysis. Right figure: A figure showing the measured values of hsa-miR-6893-5p (sequence number 2) of healthy subjects (50 people, circles) and biliary tract cancer patients (33 people, triangles) selected as the test sample group as the horizontal axis and the measured values of hsa-miR-4476 (sequence number 4) as the vertical axis. The line in the figure represents the threshold value (0=5.16x+y+48.11) for distinguishing the two groups, which was set based on the learning sample group.

[0285] Figure 4 Middle, upper figure: hsa-miR-6075 (SEQ ID NO. 15), hsa-miR-6836-3p (SEQ ID NO. 12), hsa-miR-6799-5p (SEQ ID NO. 29), hsa-miR-1 in 67 patients with biliary tract cancer, 93 healthy subjects, 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 learning sample group. The discriminant equation (-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) was constructed using Fisher's discriminant analysis for the measured values of 25a-3p (SEQ ID NO: 1). The discriminant score obtained using this discriminant equation is plotted on the vertical axis and the sample group on the horizontal axis. The dotted line in the figure indicates the discriminant boundary between the two groups, where the discriminant score is 0. The figure below shows the measured values of hsa-miR-6075 (SEQ ID NO: 15), hsa-miR-6836-3p (SEQ ID NO: 12), hsa-miR-6799-5p (SEQ ID NO: 29), and hsa-miR-125a-3p (SEQ ID NO: 1) for 33 patients with biliary tract cancer, 57 healthy subjects, 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, with the discriminant score obtained by the discriminant formula generated by the learning sample group as the vertical axis and the sample group as the horizontal axis. The dotted line in the figure represents the discrimination boundary between the two groups with a discrimination score of 0. DETAILED DESCRIPTION

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

[0287] 1. Target nucleic acids for biliary tract cancer

[0288] Among the main target nucleic acids used as biliary tract cancer markers for detecting the presence and / or absence of biliary tract cancer or biliary tract cancer cells using the nucleic acid probe or primer for biliary tract cancer detection defined above, 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-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 selected from the group consisting of hsa-miR-6769b-5p, hsa-miR-4508, hsa-miR-6826-5p, hsa-miR-6757-5p, hsa-miR-3131, and hsa-miR-1343-3p. Other biliary tract cancer markers that can be combined with these miRNAs include 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 selected from the group consisting of 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 can also be preferably used as the target nucleic acid.

[0289] Among the above-mentioned miRNAs, there are human genes containing the base sequences shown in any one of, for example, 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-6 845-5p, 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-4674、hsa-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- hsa-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 homologs, their transcription products, and their mutants or derivatives. Here, genes, homologs, transcription products, mutants and derivatives are as defined above.

[0290] A preferred target nucleic acid is a human gene comprising the base sequence shown in any one of SEQ ID NOs: 1 to 509, or a transcription product thereof, more preferably the transcription product, i.e., miRNA, or its precursor RNA, pri-miRNA or pre-miRNA.

[0291] The first target gene is the hsa-miR-125a-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0292] The second target gene is the hsa-miR-6893-5p gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0293] The third target gene is the hsa-miR-204-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0294] The fourth target gene is the hsa-miR-4476 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0295] The fifth target gene is the hsa-miR-4294 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0296] The sixth target gene is the hsa-miR-150-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0297] The seventh target gene is the hsa-miR-6729-5p gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0298] The eighth target gene is the hsa-miR-7641 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0299] The ninth target gene is the hsa-miR-6765-3p gene, its homologs, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0300] The tenth target gene is the hsa-miR-6820-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0301] The 11th target gene is the hsa-miR-575 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0302] The 12th target gene is the hsa-miR-6836-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0303] The 13th target gene is the hsa-miR-1469 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0304] The 14th target gene is the hsa-miR-663a gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0305] The 15th target gene is the hsa-miR-6075 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0306] The 16th target gene is the hsa-miR-4634 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0307] The 17th target gene is the hsa-miR-423-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0308] The 18th target gene is the hsa-miR-4454 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0309] The 19th target gene is the hsa-miR-7109-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0310] The 20th target gene is the hsa-miR-6789-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0311] The 21st target gene is the hsa-miR-6877-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0312] The 22nd target gene is the hsa-miR-4792 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0313] The 23rd target gene is the hsa-miR-4530 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0314] The 24th target gene is the hsa-miR-7975 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0315] The 25th target gene is the hsa-miR-6724-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0316] The 26th target gene is the hsa-miR-8073 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0317] The 27th target gene is the hsa-miR-7977 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0318] The 28th target gene is the hsa-miR-1231 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0319] The 29th target gene is the hsa-miR-6799-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0320] The 30th target gene is the hsa-miR-615-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0321] The 31st target gene is the hsa-miR-4450 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0322] The 32nd target gene is the hsa-miR-6726-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0323] The 33rd target gene is the hsa-miR-6875-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0324] The 34th target gene is the hsa-miR-4734 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0325] The 35th target gene is the hsa-miR-16-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0326] The 36th target gene is the hsa-miR-602 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0327] The 37th target gene is the hsa-miR-4651 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0328] The 38th target gene is the hsa-miR-8069 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0329] The 39th target gene is the hsa-miR-1238-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0330] The 40th target gene is the hsa-miR-6880-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0331] The 41st target gene is the hsa-miR-8072 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0332] The 42nd target gene is the hsa-miR-4723-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0333] The 43rd target gene is the hsa-miR-4732-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0334] The 44th target gene is the hsa-miR-6125 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0335] The 45th target gene is the hsa-miR-6090 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0336] The 46th target gene is the hsa-miR-7114-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0337] The 47th target gene is the hsa-miR-564 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0338] The 48th target gene is the hsa-miR-451a gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0339] The 49th target gene is the hsa-miR-3135b gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0340] The 50th target gene is the hsa-miR-4497 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0341] The 51st target gene is the hsa-miR-4665-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0342] The 52nd target gene is the hsa-miR-3622a-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0343] The 53rd target gene is the hsa-miR-6850-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0344] The 54th target gene is the hsa-miR-6821-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0345] The 55th target gene is the hsa-miR-5100 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0346] The 56th target gene is the hsa-miR-6872-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0347] The 57th target gene is the hsa-miR-4433-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0348] The 58th target gene is the hsa-miR-1227-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0349] The 59th target gene is the hsa-miR-3188 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0350] The 60th target gene is the hsa-miR-7704 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0351] The 61st target gene is the hsa-miR-3185 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0352] The 62nd target gene is the hsa-miR-1908-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0353] The 63rd target gene is the hsa-miR-6781-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0354] The 64th target gene is the hsa-miR-6805-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0355] The 65th target gene is the hsa-miR-8089 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0356] The 66th target gene is the hsa-miR-665 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0357] The 67th target gene is the hsa-miR-4486 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0358] The 68th target gene is the hsa-miR-6722-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0359] The 69th target gene is the hsa-miR-1260a gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0360] The 70th target gene is the hsa-miR-4707-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0361] The 71st target gene is the hsa-miR-6741-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0362] The 72nd target gene is the hsa-miR-1260b gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0363] The 73rd target gene is the hsa-miR-1246 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0364] The 74th target gene is the hsa-miR-6845-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0365] The 75th target gene is the hsa-miR-4638-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0366] The 76th target gene is the hsa-miR-6085 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0367] The 77th target gene is the hsa-miR-1228-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0368] The 78th target gene is the hsa-miR-4534 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0369] The 79th target gene is the hsa-miR-5585-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0370] The 80th target gene is the hsa-miR-4741 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0371] The 81st target gene is the hsa-miR-4433b-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0372] The 82nd target gene is the hsa-miR-197-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0373] The 83rd target gene is the hsa-miR-718 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0374] The 84th target gene is the hsa-miR-4513 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0375] The 85th target gene is the hsa-miR-4446-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0376] The 86th target gene is the hsa-miR-619-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0377] The 87th target gene is the hsa-miR-6816-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0378] The 88th target gene is the hsa-miR-6778-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0379] The 89th target gene is the hsa-miR-24-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0380] The 90th target gene is the hsa-miR-1915-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0381] The 91st target gene is the hsa-miR-4665-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0382] The 92nd target gene is the hsa-miR-4449 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0383] The 93rd target gene is the hsa-miR-6889-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0384] The 94th target gene is the hsa-miR-486-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0385] The 95th target gene is the hsa-miR-7113-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0386] The 96th target gene is the hsa-miR-642a-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0387] The 97th target gene is the hsa-miR-7847-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0388] The 98th target gene is the hsa-miR-6768-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0389] The 99th target gene is the hsa-miR-1290 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0390] The 100th target gene is the hsa-miR-7108-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0391] The 101st target gene is the hsa-miR-92b-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0392] The 102nd target gene is the hsa-miR-663b gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0393] The 103rd target gene is the hsa-miR-3940-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0394] The 104th target gene is the hsa-miR-4467 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0395] The 105th target gene is the hsa-miR-6858-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0396] The 106th target gene is the hsa-miR-4417 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0397] The 107th target gene is the hsa-miR-3665 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0398] The 108th target gene is the hsa-miR-4736 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0399] The 109th target gene is the hsa-miR-4687-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0400] The 110th target gene is the hsa-miR-1908-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0401] The 111th target gene is the hsa-miR-5195-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0402] The 112th target gene is the hsa-miR-4286 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0403] The 113th target gene is the hsa-miR-3679-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0404] The 114th target gene is the hsa-miR-6791-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0405] The 115th target gene is the hsa-miR-1202 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0406] The 116th target gene is the hsa-miR-3656 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0407] The 117th target gene is the hsa-miR-4746-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0408] The 118th target gene is the hsa-miR-3184-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0409] The 119th target gene is the hsa-miR-3937 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0410] The 120th target gene is the hsa-miR-6515-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0411] The 121st target gene is the hsa-miR-6132 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0412] The 122nd target gene is the hsa-miR-187-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0413] The 123rd target gene is the hsa-miR-7111-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0414] The 124th target gene is the hsa-miR-5787 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0415] The 125th target gene is the hsa-miR-6779-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0416] The 126th target gene is the hsa-miR-6808-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0417] The 127th target gene is the hsa-miR-6774-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0418] The 128th target gene is the hsa-miR-4656 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0419] The 129th target gene is the hsa-miR-6806-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0420] The 130th target gene is the hsa-miR-1233-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0421] The 131st target gene is the hsa-miR-328-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0422] The 132nd target gene is the hsa-miR-4674 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0423] The 133rd target gene is the hsa-miR-2110 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0424] The 134th target gene is the hsa-miR-6076 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0425] The 135th target gene is the hsa-miR-3619-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there have been no reports that changes in the expression of this gene or its transcripts can serve as markers for biliary tract cancer.

[0426] The 136th target gene is the hsa-miR-92a-2-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0427] The 137th target gene is the hsa-miR-128-1-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0428] The 138th target gene is the hsa-miR-638 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0429] The 139th target gene is the hsa-miR-2861 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0430] The 140th target gene is the hsa-miR-371a-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0431] The 141st target gene is the hsa-miR-211-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0432] The 142nd target gene is the hsa-miR-1273g-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0433] The 143rd target gene is the hsa-miR-1203 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0434] The 144th target gene is the hsa-miR-122-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0435] The 145th target gene is the hsa-miR-4258 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0436] The 146th target gene is the hsa-miR-4484 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0437] The 147th target gene is the hsa-miR-4648 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0438] The 148th target gene is the hsa-miR-6780b-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of this gene or its transcripts can serve as a marker for biliary tract cancer.

[0439] The 149th target gene is the hsa-miR-4516 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0440] The 150th target gene is the hsa-miR-4649-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0441] The 151st target gene is the hsa-miR-760 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0442] The 152nd target gene is the hsa-miR-3162-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0443] The 153rd target gene is the hsa-miR-3178 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0444] The 154th target gene is the hsa-miR-940 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0445] The 155th target gene is the hsa-miR-4271 gene, its homologs, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0446] The 156th target gene is the hsa-miR-6769b-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0447] The 157th target gene is the hsa-miR-4508 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0448] The 158th target gene is the hsa-miR-6826-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0449] The 159th target gene is the hsa-miR-6757-5p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0450] The 160th target gene is the hsa-miR-3131 gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

[0451] The 161st target gene is the hsa-miR-1343-3p gene, its homologues, its transcription products, or its mutants or derivatives. To date, there has been no report that changes in the expression of genes or their transcripts can serve as markers for biliary tract cancer.

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

[0453] In the present invention, a nucleic acid that can specifically bind to the above-mentioned target nucleic acid serving as a biliary tract cancer marker can be used as a nucleic acid for detecting or diagnosing biliary tract cancer, such as a nucleic acid probe or primer.

[0454] In the present invention, the nucleic acid probe or primer that can be used to detect biliary tract cancer or can be used to diagnose biliary tract cancer can qualitatively and / or quantitatively measure the target nucleic acid as the above-mentioned biliary tract cancer marker, such as 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 a combination thereof, or a homologue thereof, a transcription product thereof, a mutant or derivative thereof, 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 presence, expression or presence 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 a combination thereof, or a homologue thereof, a transcription product thereof, a mutant or derivative thereof.

[0455] Regarding the above-mentioned target nucleic acids, compared with healthy bodies, in subjects suffering from biliary tract cancer, depending on the type of the target nucleic acid, their expression levels are both increased and decreased (hereinafter referred to as "increase / decrease."). Therefore, the nucleic acid of the present invention can measure the expression levels of the above-mentioned target nucleic acids in body fluids derived from subjects (such as humans) suspected of suffering from biliary tract cancer and body fluids derived from healthy bodies, and compare them, which is effectively used to detect biliary tract cancer. In addition, the nucleic acid of the present invention can measure the expression levels of the above-mentioned target nucleic acids in body fluids derived from subjects (such as humans) suspected of suffering from biliary tract cancer, as well as body fluids derived from patients with colorectal cancer, gastric cancer, esophageal cancer, liver cancer, and benign pancreatic and biliary tract diseases, and compare them, which is effectively used to specifically detect biliary tract cancer from other cancers, benign diseases, etc.

[0456] The nucleic acid probe or primer that can be used in the present invention is a nucleic acid probe that can specifically bind to a polynucleotide consisting of a base sequence represented by at least one of sequence numbers 1 to 125 (preferably sequence numbers 1, 2, 4 to 125) and 466 to 478, or a primer for amplifying a polynucleotide consisting of a base sequence represented by at least one of sequence numbers 1 to 125 and 466 to 478.

[0457] The nucleic acid probe or primer that can be used in the present invention may further include a nucleic acid probe that can specifically bind to a polynucleotide consisting of at least one base sequence represented by sequence numbers 126 to 148, or a primer for amplifying a polynucleotide consisting of at least one base sequence represented by sequence numbers 126 to 148.

[0458] Specifically, the nucleic acid probe or primer comprises a polynucleotide group selected from a base sequence shown in any one of SEQ ID NOs. 1 to 509 or a base sequence in which u is t in the base sequence, and a complementary polynucleotide group thereof, a polynucleotide group and a complementary polynucleotide group that hybridize under stringent conditions (described later) with a DNA consisting of a base sequence complementary to the base sequence, and a polynucleotide group comprising 15 or more, preferably 17 or more, consecutive bases in the base sequence of these polynucleotide groups, and a combination of one or more polynucleotides. These polynucleotides can be used as nucleic acid probes and primers for detecting the above-mentioned biliary tract cancer marker as a target nucleic acid.

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

[0460] (a) a polynucleotide consisting of the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0461] (b) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478,

[0462] (c) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0463] (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, and

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

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

[0466] (f) a polynucleotide consisting of the base sequence represented by any of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0467] (g) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 126 to 148,

[0468] (h) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0469] (i) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, and

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

[0471] The "fragment of the polynucleotide comprising 15 or more consecutive bases" in the above-mentioned polynucleotide may, in the base sequence of each polynucleotide, include, for example, a consecutive number of bases ranging from 15 to less than the total number of bases in the sequence, from 17 to less than the total number of bases in the sequence, from 19 to less than the total number of bases in the sequence, etc., but is not limited thereto.

[0472] The polynucleotides or fragments thereof used in the present invention may be DNA or RNA.

[0473] The polynucleotides that can be used in the present invention can be produced using common techniques such as DNA recombination technology, PCR, and methods using a DNA / RNA automatic synthesizer.

[0474] For DNA recombination technology and PCR method, for example, the techniques described in 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) and the like can be used.

[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-m iR-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-3619-3p、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, and hsa-miR-6780b-5p are known, and methods for obtaining them are also known as described above. Therefore, by cloning these genes, polynucleotides that can be used as nucleic acid probes or primers in the present invention can be prepared.

[0476] Such nucleic acid probes or primers can be chemically synthesized using an automated DNA synthesizer. This synthesis generally uses the phosphoramidite method, which allows the automatic synthesis of single-stranded DNA of up to about 100 bases. Automated DNA synthesizers are commercially available from companies such as Polygen, ABI, and Applied BioSystems.

[0477] Alternatively, the polynucleotide of the present invention can also be prepared by cDNA cloning. For example, the cDNA cloning technique can utilize the microRNA Cloning Kit (Wako).

[0478] Here, the sequences of nucleic acid probes and primers for detecting a polynucleotide consisting of a base sequence represented by any one of SEQ ID NOs: 1 to 148 and 466 to 478 do not exist in vivo as miRNA or its precursor. For example, the base sequences represented by SEQ ID NOs: 51 and 91 are generated from the precursor represented by SEQ ID NO: 201, but the precursor has Figure 1 In the hairpin-like structure shown, the base sequences shown in SEQ ID NO: 51 and SEQ ID NO: 91 have mismatches. Therefore, base sequences completely complementary to the base sequences shown in SEQ ID NO: 51 or SEQ ID NO: 91 do not naturally occur in living organisms. Similarly, nucleic acid probes and primers used to detect the base sequences shown in any of SEQ ID NOs: 1 to 148 and 466 to 478 have artificial base sequences that do not exist in living organisms.

[0479] 3. Kits or devices for detecting biliary tract cancer

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

[0481] The target nucleic acid used as a biliary tract cancer marker in the present invention is preferably selected from the following group 1:

[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 situation, the additional target nucleic acids that can be used for determination 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 that can specifically bind to the above-mentioned target nucleic acid as a biliary cancer marker, preferably contains the nucleic acid probe or primer described in 2 above, specifically contains one or more polynucleotides selected from the polynucleotide types described in 2 above or mutants of the polynucleotide, etc.

[0485] Specifically, the kit or device of the present invention may contain at least one or more of the following: a polynucleotide comprising a base sequence represented by any one of SEQ ID NOs. 1 to 125 and 466 to 478 or a base sequence in which u is t in the base sequence (or consisting of a base sequence represented by any one of SEQ ID NOs. 1 to 125 and 466 to 478 or a base sequence in which u is t in the base sequence), a polynucleotide comprising a complementary sequence thereof (or consisting of a complementary sequence thereof), a polynucleotide that hybridizes to these polynucleotides under stringent conditions, or a mutant or fragment of these polynucleotide sequences comprising 15 or more consecutive bases.

[0486] The kit or device of the present invention may further include one or more of the following: a polynucleotide comprising a base sequence represented by any one of sequence numbers 126 to 148 or a base sequence in which u is t in the base sequence (or consisting of a base sequence represented by any one of sequence numbers 126 to 148 or a base sequence in which u is t in the base sequence), a polynucleotide comprising a complementary sequence thereof (or consisting of a complementary sequence thereof), a polynucleotide that hybridizes to these polynucleotides under stringent conditions, or a mutant or fragment of these polynucleotide sequences comprising more than 15 consecutive bases.

[0487] The fragments that can be contained in the kit or device of the present invention are, for example, one or more, preferably two or more, polynucleotides selected from the following (1) to (2).

[0488] (1) A polynucleotide comprising 15 or more consecutive bases in a base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, wherein u is t, or a complementary sequence thereof.

[0489] (2) A polynucleotide comprising 15 or more consecutive bases in a base sequence in which u is t in any one of SEQ ID NOs: 126 to 148 or a complementary sequence thereof.

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

[0491] In addition, in a preferred embodiment, the above-mentioned polynucleotide is a polynucleotide composed of a base sequence shown in any one of sequence numbers 126 to 148 or a base sequence in which u is t in the base sequence; a polynucleotide composed of its complementary sequence; a polynucleotide that hybridizes with these polynucleotides under stringent conditions; or a mutant thereof containing more than 15, preferably more than 17, and more preferably more than 19 consecutive bases.

[0492] In a preferred embodiment, the fragment may be a polynucleotide comprising 15 or more, preferably 17 or more, more preferably 19 or more consecutive bases.

[0493] In the present invention, the fragment size of a polynucleotide is, for example, a continuous number of bases ranging from 15 to less than the total number of bases in the base sequence of each polynucleotide, from 17 to less than the total number of bases in the sequence, from 19 to less than the total number of bases in the sequence, etc.

[0494] As a combination of the above-mentioned polynucleotides constituting the kit or device of the present invention, specifically, any combination of the above-mentioned polynucleotides consisting of the base sequences represented by the sequence numbers shown in Table 1 below (sequence numbers 1 to 148 and 466 to 478 corresponding to the miRNA markers in Table 1) or their complementary sequences can be cited. 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-mentioned combination constituting a kit or device for distinguishing biliary tract cancer from healthy bodies, it is expected that the above-mentioned polynucleotides composed of the base sequence represented by the sequence number shown in Table 1 are combined in two or more. Usually, a combination of two can obtain sufficient performance.

[0496] Specifically, as a combination of two polynucleotides composed of a base sequence or its complementary sequence for distinguishing biliary tract cancer from healthy bodies, among the combinations of two of the above-mentioned polynucleotides composed of the base sequences shown in sequence numbers 1 to 148 and 466 to 478, it is preferred to include a combination of at least one newly discovered polynucleotide composed of the base sequences shown in sequence numbers 1 to 125 and 466 to 478.

[0497] In addition, as a combination of cancer type-specific polynucleotides that can distinguish not only healthy people from biliary tract cancer but also healthy people from other cancers, it is preferred to select, for example, at least one polynucleotide from a group consisting of polynucleotides consisting of 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 referred to as "cancer type-specific polynucleotide group 1"), and a combination of multiple polynucleotides of other sequence numbers.

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

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

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

[0501] The following non-limiting examples include combinations of a polynucleotide consisting of the base sequence represented by SEQ ID NO: 4 or its complementary sequence, and a polynucleotide consisting of the base sequences represented by the sequence numbers of three polynucleotides selected from cancer type-specific polynucleotide group 1 or their complementary sequences.

[0502] (1) Combination of SEQ ID NOs: 4, 15, 54, and 115 (markers: miR-4476, miR-6075, miR-6821-5p, and miR-1202)

[0503] (2) Combination of SEQ ID NOs: 4, 5, 12, and 76 (markers: miR-4476, miR-4294, miR-6836-3p, and miR-6085)

[0504] (3) Combination of SEQ ID NOs: 4, 5, 12, and 115 (markers: miR-4476, miR-4294, miR-6836-3p, and miR-1202)

[0505] (4) Combination of SEQ ID NOs: 4, 12, 15, and 474 (markers: miR-4476, miR-6836-3p, miR-6075, and miR-4508)

[0506] (5) Combination of SEQ ID NOs: 4, 15, 29, and 115 (markers: miR-4476, miR-6075, miR-6799-5p, and miR-1202)

[0507] The following non-limiting examples include combinations of a polynucleotide consisting of the base sequence represented by SEQ ID NO: 5 or its complementary sequence, and a polynucleotide consisting of the base sequences represented by the sequence numbers of three polynucleotides selected from cancer type-specific polynucleotide group 1 or their complementary sequences.

[0508] (1) Combination of SEQ ID NOs: 5, 76, 12, and 115 (markers: hsa-miR-4294, hsa-miR-6085, hsa-miR-6836-3p, and hsa-miR-1202)

[0509] (2) Combination of SEQ ID NOs: 5, 76, 54, and 115 (markers: hsa-miR-4294, hsa-miR-6085, hsa-miR-6821-5p, and hsa-miR-1202)

[0510] (3) Combination of SEQ ID NOs: 5, 23, 12, and 115 (markers: hsa-miR-4294, hsa-miR-4530, hsa-miR-6836-3p, and hsa-miR-1202)

[0511] (4) Combination of SEQ ID NOs: 5, 12, 115, and 91 (markers: hsa-miR-4294, hsa-miR-6836-3p, hsa-miR-1202, and hsa-miR-4665-3p)

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

[0513] The following non-limiting examples include combinations of a polynucleotide consisting of the base sequence represented by SEQ ID NO: 12 or its complementary sequence and a polynucleotide consisting of the base sequences represented by the sequence numbers of three polynucleotides selected from cancer type-specific polynucleotide group 1 or their complementary sequences.

[0514] (1) Combination of SEQ ID NOs: 5, 12, 29, and 115 (markers: miR-4294, miR-6836-3p, miR-6799-5p, and miR-1202)

[0515] (2) Combination of SEQ ID NOs: 12, 15, 23, and 115 (markers: miR-6836-3p, miR-6075, miR-4530, and miR-1202)

[0516] (3) Combination of SEQ ID NOs: 5, 12, 115, and 469 (markers: miR-4294, miR-6836-3p, miR-3162-5p, and miR-1202)

[0517] (4) Combination of SEQ ID NOs: 5, 12, 115, and 472 (markers: miR-4294, miR-6836-3p, miR-1202, and miR-4271)

[0518] (5) Combination of SEQ ID NOs: 5, 12, 76, and 115 (markers: miR-4294, miR-6085, miR-1202, and miR-6836-3p)

[0519] The following non-limiting examples include combinations of a polynucleotide consisting of the base sequence represented by SEQ ID NO: 15 or its complementary sequence and a polynucleotide consisting of the base sequences represented by the sequence numbers of three polynucleotides selected from cancer type-specific polynucleotide group 1 or their complementary sequences.

[0520] (1) Combination of SEQ ID NOs: 15, 29, 1, and 12 (markers: hsa-miR-6075, hsa-miR-6799-5p, hsa-miR-125a-3p, and hsa-miR-6836-3p)

[0521] (2) Combination of SEQ ID NOs: 15, 12, 11, and 143 (markers: hsa-miR-6075, hsa-miR-6836-3p, hsa-miR-575, and hsa-miR-1203)

[0522] (3) Combination of SEQ ID NOs: 15, 76, 121, and 39 (markers: hsa-miR-6075, hsa-miR-6085, hsa-miR-6132, and hsa-miR-1238-5p)

[0523] (4) Combination of SEQ ID NOs: 15, 76, 54, and 121 (markers: hsa-miR-6075, hsa-miR-6085, hsa-miR-6821-5p, and hsa-miR-6132)

[0524] (5) Combination of SEQ ID NOs: 15, 40, 1, and 23 (markers: hsa-miR-6075, hsa-miR-6880-5p, hsa-miR-125a-3p, and hsa-miR-4530)

[0525] The following non-limiting examples include combinations of a polynucleotide consisting of the base sequence represented by SEQ ID NO: 40 or its complementary sequence and a polynucleotide consisting of the base sequences represented by the sequence numbers of three polynucleotides selected from cancer type-specific polynucleotide group 1 or their complementary sequences.

[0526] (1) Combination of SEQ ID NOs: 12, 40, 472, and 473 (markers: miR-6836-3p, miR-6880-5p, miR-4271, and miR-6769b-5p)

[0527] (2) Combination of SEQ ID NOs: 12, 23, 40, and 466 (markers: miR-6836-3p, miR-4530, miR-6880-5p, and miR-4516)

[0528] (3) Combination of SEQ ID NOs: 12, 23, 40, and 134 (markers: miR-6836-3p, miR-4530, miR-6880-5p, and miR-6076)

[0529] (4) Combination of SEQ ID NOs: 15, 40, 121, and 134 (markers: miR-6075, miR-6880-5p, miR-6132, and miR-6076)

[0530] (5) Combination of SEQ ID NOs: 15, 40, 54, and 76 (markers: miR-6075, miR-6880-5p, miR-6821-5p, and miR-6085)

[0531] The kit or device of the present invention may include, in addition to the polynucleotides of the present invention described above (which may include mutants, fragments or derivatives), known polynucleotides capable of detecting biliary tract cancer or polynucleotides to be discovered in the future.

[0532] The kit of the present invention may also include, in addition to the polynucleotides of the present invention described above, antibodies for measuring well-known markers for biliary tract cancer examination, such as CEA, CA19-9, SPan-1, DUPAN-2, CA50, CA195, IL-6, CA242, TAG-72, urinary fucose, POA, and TPS.

[0533] The above-mentioned polynucleotides contained in the kit of the present invention may be packaged in different containers individually or in any combination.

[0534] The kit of the present invention may further include a kit for extracting nucleic acid (eg, total RNA) from body fluids, cells, or tissues, a fluorescent substance for labeling, an enzyme and culture medium for nucleic acid amplification, instructions for use, and the like.

[0535] The device of the present invention is a device for measuring cancer markers, wherein the nucleic acids such as polynucleotides in the present invention described above are, for example, cancer markers bound to or attached to a solid phase. Examples of materials for the solid phase are plastic, paper, glass, silicon, etc., and from the perspective of ease of processing, the preferred material for the solid phase is plastic. The shape of the solid phase is arbitrary, for example, square, circular, long strip, membrane-shaped, etc. The device of the present invention includes, for example, a device for measuring by hybridization technology, specifically, blotting devices, nucleic acid arrays (such as microarrays, DNA chips, RNA chips, etc.), etc.

[0536] Nucleic acid array technology is a technology that uses the following methods to produce an array such as a chip by binding or attaching the above-mentioned nucleic acids one by one, and uses this array to measure target nucleic acids through hybridization. The methods include: methods of spotting nucleic acids on the surface of a solid phase that has been subjected to surface treatments such as L-lysine coating and / or introduction of functional groups such as amino groups and carboxyl groups as needed, using a high-density dispenser called a spotter or arrayer; methods of spraying nucleic acids onto a solid phase using an inkjet that ejects tiny droplets from a nozzle using a piezoelectric element; methods of sequentially synthesizing nucleotides on the solid phase, etc.

[0537] The kit or device of the present invention comprises a nucleic acid 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 in Group 1 as biliary tract cancer markers. Depending on the circumstances, the kit or device of the present invention may further comprise a nucleic acid 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 in Group 2 as biliary tract cancer markers.

[0538] The kit or device of the present invention can be used for the detection of the following 4 biliary tract cancers.

[0539] 4. Detection methods for biliary tract cancer

[0540] The present invention further provides a method for detecting biliary tract cancer, which comprises: using the kit or device of the present invention described in 3. above (containing the above-mentioned nucleic acid that can be used in the present invention), in vitro measuring the following groups selected from the group 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 in the specimen. ,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 biliary tract cancer-derived genes iR-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 the expression levels of genes selected from the group consisting of 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,miR-6780b-5p,miR-4516,miR-4649-5p,miR-760,miR-3162-5p,miR-3178,m The expression levels of the biliary tract cancer-derived genes 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 the biliary tract cancer-derived genes shown in more than one of the above, and further, regarding blood, serum, plasma and other specimens collected from subjects suspected of having biliary tract cancer and healthy subjects (including non-biliary tract cancer patients), the expression levels of the above genes in the specimens and the control expression levels of the healthy subjects are used, for example, to compare the two expression levels. If there is a statistically significant difference in the expression level of the target nucleic acid in the specimen, it is evaluated that the subject has biliary tract cancer.

[0541] The method of the present invention enables low-invasive early diagnosis of cancer with high sensitivity and specificity, thereby leading to early treatment and improved prognosis, and further enables monitoring of disease progression and the effectiveness of surgical treatment, radiotherapy, and chemotherapy.

[0542] For the method of extracting genes derived from biliary tract cancer from specimens such as blood, serum, and plasma of the present invention, it is particularly preferred to add and prepare the RNA extraction reagent in the 3D-Gene (registered trademark) RNA extraction reagent from liquid sample kit (Toray Industries, Inc.), but the general acid phenol method (acid guanidinium-phenol-chloroform (AGPC) method) can also be used, Trizol (registered trademark) (Life Technologies Co., Ltd.) can also be used, and RNA extraction reagents containing acid phenol such as Trizol (life technologies company) and Isogen (Nippon Gene Co., Ltd.) can also be added and prepared. Further, kits such as miRNeasy (registered trademark) Mini Kit (Qiagen Co., Ltd.) can be used, but are not limited to these methods.

[0543] The present invention further provides the use of the kit or device of the present invention for in vitro detection of expression products of miRNA genes derived from biliary tract cancer in a specimen derived from a subject.

[0544] In the above method of the present invention, the above kit or device contains the polynucleotides that can be used in the present invention, as described above, either singly or in all possible combinations.

[0545] In the detection or (gene) 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, Life Technologies' TaqMan (registered trademark) MicroRNA Assays, Qiagen's miScript PCR System, etc. can be used, but are not limited to these methods.

[0546] The polynucleotides contained in the kits or devices of the present invention can be used as primers or probes in conventional methods for specifically detecting specific genes, such as hybridization techniques such as Northern blotting, Southern blotting, in situ hybridization, Northern hybridization, and Southern hybridization, and quantitative amplification techniques such as quantitative RT-PCR. As the test specimen, body fluids such as blood, serum, plasma, and urine are collected from the subject, depending on the type of detection method used. Alternatively, total RNA prepared from such body fluids using the above-described methods can be used, and further, various polynucleotides including cDNA prepared based on this RNA can be used.

[0547] The kit or device of the present invention is useful for diagnosing biliary tract cancer or detecting the presence or absence of biliary tract cancer. Specifically, the detection of biliary tract cancer using the kit or device can be performed as follows: from a subject suspected of suffering from biliary tract cancer, using blood, serum, plasma, urine and other specimens, in vitro detection of the expression level of the gene detected by the nucleic acid probe or primer contained in the kit or device. When the expression level of the target miRNA marker measured by the polynucleotide composed of the base sequence shown by at least one of sequence numbers 1 to 125, 466 to 478 or its complementary sequence, and the base sequence shown by one or more of sequence numbers 126 to 148 or its complementary sequence in the blood, serum, plasma, urine and other specimens of the subject suspected of suffering from biliary tract cancer is statistically significantly different from their expression levels in the blood, serum, plasma, urine and other specimens of a healthy body, it can be evaluated that the subject is suffering from biliary tract cancer.

[0548] The method of the present invention can be combined with imaging diagnostic methods such as abdominal ultrasound, CT scan, endoscopic retrograde cholangiopancreatography, and endoscopic ultrasound. The method of the present invention can specifically detect biliary tract cancer and can substantially distinguish it from cancers other than biliary tract cancer. In particular, in the case of pancreatic cancer, some common miRNA markers can be used as in the case of biliary tract cancer. However, biliary tract cancer and pancreatic cancer can be distinguished by using the discriminant boundary method of the discriminant, or these cancers can be distinguished by combining with other diagnostic methods such as the above-mentioned imaging diagnostic methods.

[0549] The method for detecting that the expression product of a gene derived from biliary tract cancer is not contained in a specimen using the kit or device of the present invention, or that the expression product of a gene derived from biliary tract cancer is contained, comprises: collecting body fluids such as blood, serum, plasma, and urine from the subject, and using a single or multiple polynucleotides (including mutants, fragments, or derivatives) selected from the polynucleotide group of the present invention to measure the expression level of the target gene contained therein, thereby evaluating the presence or absence of biliary tract cancer or detecting biliary tract cancer. In addition, the method for detecting biliary tract cancer of the present invention can also be used to evaluate or diagnose, for example, the presence or absence of improvement in the disease or the degree of improvement in a patient with biliary tract cancer who has been administered a therapeutic drug for improving the disease.

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

[0551] (a) a step of contacting a sample derived from a subject with a polynucleotide in the kit or device of the present invention in vitro,

[0552] (b) a step of measuring the expression level of a target nucleic acid in a sample using the polynucleotide as a nucleic acid probe or primer,

[0553] (c) A step of evaluating the presence or absence of biliary tract cancer (cells) in the subject based on the result of (b).

[0554] Specifically, the present invention provides a method for detecting biliary tract cancer, which includes: using a substance capable of binding to a selected substance 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-4449,miR-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 invention relates to a method for detecting a nucleic acid that specifically binds to at least one, preferably at least two, polynucleotide of 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, and measuring the expression level of the target nucleic acid in the specimen of the subject. The measured expression level and the control expression level of a healthy body measured in the same manner are used to perform in vitro evaluation of whether the subject has biliary tract cancer or not. 、

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

[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, and miR-7109-5p is hsa-miR-7109-5p. 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, and 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, and 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 is hsa-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, 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, and miR-6778-5p is hsa-miR-6779-5p. 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, and 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, 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, and miR-4649-5p is hsa-miR-4649-5p. 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, a probe or a primer) is selected from the polynucleotides shown in the following (a) to (e),

[0558] (a) a polynucleotide consisting of the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0559] (b) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478,

[0560] (c) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0561] (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 1 to 125 and 466 to 478, or a base sequence in which u is t in the base sequence, and

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

[0563] In the method of the present invention, a 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- A nucleic acid that specifically binds to at least one of the polynucleotides selected from the group consisting of 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, 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 hsa-miR-328-5p, miR-4674 is hsa-miR-4674, miR-2110 is 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, miR- 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, and, 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-3162-5p. R-3162-5p, miR-3178 was hsa-miR-3178, miR-940 was hsa-miR-940, miR-4271 was hsa-miR-4271, miR-6769b-5p was hsa-miR-6769b-5p, miR-4508 was hsa-miR-4508, miR-6826-5p was hsa-miR-6826-5p, miR-6757-5p was hsa-miR-6757-5p, miR-3131 was hsa-miR-3131, and miR-1343-3p was hsa-miR-1343-3p.

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

[0566] (f) a polynucleotide consisting of the base sequence represented by any of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a mutant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0567] (g) a polynucleotide comprising the base sequence represented by any one of SEQ ID NOs: 126 to 148,

[0568] (h) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, a variant of the polynucleotide, a derivative of the polynucleotide, or a fragment of the polynucleotide comprising 15 or more consecutive bases,

[0569] (i) a polynucleotide comprising a base sequence complementary to the base sequence represented by any one of SEQ ID NOs: 126 to 148, or a base sequence in which u is t in the base sequence, and

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

[0571] Examples of specimens used in the methods of the present invention include specimens prepared from a subject's living tissue (preferably biliary tissue), blood, serum, plasma, urine, and other body fluids. Specifically, specimens containing RNA prepared from such tissues; specimens containing polynucleotides further prepared from such tissues; and body fluids such as blood, serum, plasma, and urine; and specimens prepared from a portion or all of a subject's living tissue collected by biopsy or surgical removal.

[0572] In this specification, the subject refers to a mammal, for example, but not limited to, a human, a monkey, a mouse, a rat, etc., preferably a human.

[0573] The method of the present invention can change the steps depending on the type of sample used as the measurement object.

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

[0575] (a) a step of combining RNA prepared from a sample of a subject or a complementary polynucleotide (cDNA) transcribed therefrom with the polynucleotide in the kit or device of the present invention,

[0576] (b) a step of measuring sample-derived RNA bound to the polynucleotide or cDNA synthesized from the RNA by hybridization using the polynucleotide as a nucleic acid probe or quantitative RT-PCR using the polynucleotide as a primer,

[0577] (c) A step of evaluating the presence or absence of biliary tract cancer (expression of the derived gene) based on the measurement results of the above (b).

[0578] In order to detect, examine, evaluate or diagnose biliary tract cancer (expression of genes derived therefrom) in vitro using the present invention, various hybridization methods can be used, for example. Among such hybridization methods, Northern blotting, Southern blotting, RT-PCR, DNA chip analysis, in situ hybridization, Northern hybridization, Southern hybridization, etc. can be used.

[0579] In the case of using the Northern blotting method, the presence or absence of expression of each gene in RNA and the expression level thereof can be detected and measured by using the above-mentioned nucleic acid probes that can be used in the present invention. Specifically, the following method can be exemplified: the nucleic acid probe (complementary chain) is stained with a radioactive isotope ( 32 P. 33 P. 35 S, etc.), fluorescent substances, etc., and after hybridizing it with RNA derived from the living tissue of the subject transferred to a nylon membrane, etc. according to a conventional method, a radiation detector (such as BAS-1800II (Fuji Photo Film Co., Ltd.) or a fluorescence detector (such as STORM 865 (GE Healthcare)) is used to detect and measure the signal derived from the label (radioactive isotope or fluorescent substance) of the formed DNA / RNA double chain.

[0580] When utilizing quantitative RT-PCR, the presence and level of gene expression in RNA can be detected and measured using the primers described above that can be used in the present invention. Specifically, the following methods can be exemplified: cDNA is prepared from RNA derived from a living tissue of a subject according to conventional methods, used as a template to amplify the target gene region, a pair of primers of the present invention (composed of a positive strand and a negative strand that bind to the cDNA) are hybridized with the cDNA, PCR is performed according to conventional methods, and the resulting double-stranded DNA is detected. Other methods for detecting double-stranded DNA include: performing the aforementioned PCR using primers previously labeled with a radioisotope or fluorescent substance; performing electrophoresis of the PCR product on an agarose gel and staining the double-stranded DNA with ethidium bromide or the like for detection; and transferring the resulting double-stranded DNA to a nylon membrane or the like according to conventional methods and hybridizing it with a labeled nucleic acid probe for detection.

[0581] In the case of utilizing nucleic acid array analysis, the RNA chip or DNA chip in which the nucleic acid probe (single-stranded or double-stranded) of the present invention is adhered to a substrate (solid phase) is used. The area adhered with the nucleic acid probe is referred to as a probe spot, and the area to which the nucleic acid probe is not adhered is referred to as a blank spot. In the material obtained by solid-phase immobilization of the gene group on a substrate, there are generally such names as nucleic acid chip, nucleic acid array, microarray, etc., and DNA or RNA arrays include DNA or RNA macroarrays and DNA or RNA microarrays, but in the case of mentioning the chip in this specification, all of these arrays are included. As a DNA chip, 3D-Gene (registered trademark) Human miRNA Oligo chip (Toray Industries, Inc.) can be used, but is not limited thereto.

[0582] The measurement method using a DNA chip is not limited. For example, a method in which a signal derived from a label of a nucleic acid probe is detected and measured using an image detector (for example, Typhoon 9410 (GE Healthcare), 3D-Gene (registered trademark) scanner (Toray Industries, Ltd.) etc.) can be exemplified.

[0583] As used herein, "stringent conditions" are conditions under which a nucleic acid probe hybridizes to a target sequence to a greater extent (e.g., mean background measurement value + standard error of background measurement value × 2 or more) than to other sequences as described above.

[0584] Stringent conditions are defined by the conditions for hybridization and subsequent washing. Hybridization conditions are not limited, but examples include conditions at 30°C to 60°C for 1 to 24 hours in a solution containing SSC, a surfactant, formamide, dextran sulfate, a blocking agent, and the like. 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. Hybridization conditions preferably contain 3× to 10×SSC and 0.1% SDS. Post-hybridization washing conditions, another condition defining stringent conditions, include, for example, sequential 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 preferably maintains hybridization with the target positive chain even under these washing conditions. Specifically, examples of such complementary chains include chains composed of base sequences that are completely complementary to the base sequence of the positive chain of the object, and chains composed of base sequences that are 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% identical to the positive chain.

[0585] Other examples of "stringent conditions" in these hybridizations are described in, for example, Sambrook, J. & Russell, D., Molecular Cloning, A LABORATORY MANUAL, Cold Spring Harbor Laboratory Press, published on January 15, 2001, Vol. 1, pp. 7.42 to 7.45, and Vol. 2, pp. 8.9 to 8.17, and can be used in the present 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 to 2 mM MgCl2, and performing treatment at a temperature of 5 to 10°C (the Tm value calculated from the sequence of the primer) for approximately 15 seconds to 1 minute. Examples of methods for calculating such Tm values include: Tm value = 2 × (number of adenine residues + number of thymine residues) + 4 × (number of guanine residues + number of cytosine residues).

[0587] When using quantitative RT-PCR, commercially available assay kits specifically developed for quantitative miRNA measurement, such as TaqMan (registered trademark) MicroRNA Assays (Life Technologies), LNA (registered trademark)-based MicroRNA PCR (Exiqon), and Ncode (registered trademark) miRNA qRT-PCR Kit (Invitrogen), can be used.

[0588] The calculation of gene expression level is not limited. In the present invention, the statistical processing described in, for example, Statistical analysis of gene expression microarray data (Speed T., Chapman and Hall / CRC) and A beginner's guide Microarray gene expression data analysis (Causton HC, etc., Blackwell publishing) can be utilized. For example, the mean value of the measured value of the blank spot on the DNA chip can be added with 2 times, preferably 3 times, and more preferably 6 times, of the standard deviation of the measured value of the blank spot, and the probe spot with a signal value above this value can be regarded as a detection point. Further, the mean value of the measured value of the blank spot can be regarded as background, subtracted from the measured value of the probe spot, and used as the gene expression level. For the missing value of the gene expression level, it can be removed from the analysis object, or preferably replaced with the minimum value of the gene expression level in each DNA chip, or more preferably replaced with the value after subtracting 0.1 from the logarithm of the minimum value of the gene expression level. Furthermore, in order to remove genes with low signals, only genes whose expression levels are greater than or equal to 2 to the power of 6, preferably greater than or equal to 2 to the power of 8, and more preferably greater than or equal to 2 to the power of 10 in 20% or more, preferably greater than or equal to 50%, and more preferably greater than or equal to 80% of the number of samples measured can be selected as analysis targets. Normalization of gene expression levels is not limited, and examples thereof include global normalization and quantile normalization (Bolstad, BM et al., 2003, Bioinformatics, Vol. 19, pp. 185-193).

[0589] The present invention also provides the following method: using the detection polynucleotide, kit, device (e.g., chip) of the present invention, or a combination thereof, the expression level of the target gene or genes in a specimen derived from a subject is measured, and the gene expression levels of specimens derived from biliary tract cancer patients and specimens derived from healthy subjects are used 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 the following method, which includes the following steps: a first step of in vitro measuring the expression level of a target gene (target nucleic acid) in a plurality of specimens known to contain a gene derived from biliary tract cancer or not contain a gene derived from biliary tract cancer using the detection polynucleotide, kit, device (for example, a chip) of the present invention, or a combination thereof; a second step of making a discriminant formula using the measured value of the expression level of the target gene obtained in the above-mentioned first step as the teacher sample; a third step of in vitro measuring the expression level of the target gene in the specimen derived from the subject in the same manner as in the first step; a fourth step of determining or evaluating whether the specimen contains a gene derived from biliary tract cancer / or does not contain a gene derived from biliary tract cancer based on the result obtained by 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 above-mentioned second step, and determining or evaluating whether the specimen contains a gene derived from biliary tract cancer / or does not contain a gene derived from biliary tract cancer based on the result obtained by the discriminant formula, wherein the target gene is a gene that can be detected by the detection polynucleotide contained in the polynucleotide, kit or device (for example, a chip). Here, the discriminant can be generated using Fisher's discriminant analysis, nonlinear discriminant analysis using Mahalanobis distance, neural network, support vector machine (SVM), etc., but is not limited thereto.

[0591] Linear discriminant analysis is a method for determining group membership using Equation 1 as the discriminant when the group boundary is a straight line or 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 by the discriminant equation can be called a discriminant score. The measured values of a newly given data set are substituted into the discriminant equation as explanatory variables, and the groups are discriminated based on the sign of the discriminant score.

[0595] Fisher's discriminant analysis, a type of linear discriminant analysis, is a dimensionality reduction method used to select dimensions suitable for class discrimination. It focuses on the variance of composite variables and minimizes the variance of data with the same label to construct composite variables with high discriminative power (Venables, W.N. et al., Modern Applied Statistics with S., Fourth Edition, Springer, 2002). In Fisher's discriminant analysis, the projection direction w is determined to maximize Equation 2. Here, μ is the input mean, ng is the number of data belonging to category g, and μg is the input mean of data belonging to category g. The numerator and denominator are the between-group variance and within-group variance, respectively, when the data are projected onto the direction of vector w. The discriminant coefficient wi is determined by maximizing this ratio (Kanamori Takafumi et al., "Pattern Recognition," Kyoritsu Publishing (2009), Richard O. et al., Pattern Classification, Second Edition, Wiley-Interscience, 2000).

[0596] [Number 2]

[0597]

[0598] Among them, satisfy

[0599] The Mahalanobis distance is calculated by Equation 3 which takes data correlation into account. It can be used for nonlinear discriminant analysis to discriminate groups with the closest Mahalanobis distance. Here, μ is the center vector of each group, S -1 is the inverse matrix of the variance-covariance matrix of the group. The center vector is calculated from the explanatory variable x, and the mean vector, center value vector, etc. can be used.

[0600] [Number 3]

[0601]

[0602] The so-called SVM is a discriminant analysis method studied by V. Vapnik (The Nature of Statistical Leaning Theory, Springer, 1995). The specific data items of the data set with known groups to be classified are used as explanatory variables, the groups to be classified are used as target variables, a boundary surface called a hyperplane is determined for correctly classifying the data set into the known groups, and a discriminant formula for classifying the data using the boundary surface is determined. Moreover, the discriminant formula can discriminate the groups by substituting the measured values of the newly given data set into the discriminant formula as explanatory variables. In addition, the discrimination result at this time can be the group to be classified, or the probability of being classified into the group to be classified, or the distance from the hyperplane. For SVM, as a method for dealing with nonlinear problems, a method of nonlinearly transforming the feature vector to a high dimension and performing linear discrimination in this space is known. The formula that expresses the inner product of two elements in the space of nonlinear mapping only by the input of each original space is called a kernel. As an example of a kernel, a linear kernel, an RBF (Radial Basis Function) kernel, and a Gaussian kernel can be cited. The kernel is used to map to a high dimension, and the optimal discriminant, i.e., the discriminant, is constructed only by calculation of the kernel, avoiding calculation of the features of the mapped space (for example, Hideki Aso et al., "New Concepts and Methods of Part-time Knowledge and Learning 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, uses two groups of explanatory variables to create a hyperplane and classify an unknown dataset into which group (C. Cortes et al., 1995, Machine Learning, Vol. 20, pp. 273-297).

[0604] The following is an example of calculating the discriminant of C-SVC 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 subjects. To determine whether a subject is a patient with biliary tract cancer or a healthy subject, for example, a biliary tissue examination can be used.

[0605] Next, a dataset consisting of the total gene expression levels of two separate groups of serum-derived specimens (hereinafter referred to as the learning specimen group) was prepared. A C-SVC discriminant was determined, using genes with clear differences in gene expression between the two groups as explanatory variables and the corresponding groupings as target variables (e.g., -1 and +1). Equation 4 is the optimization objective function, where e represents the total input vector, y represents the target variable, a represents the Lagrange undetermined multiplier vector, Q represents the positive definite matrix, and C represents the parameter for adjusting the constraints.

[0606] [Number 4]

[0607]

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

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

[0610] [Number 5]

[0611]

[0612] As the kernel function, for example, an RBF kernel defined by Equation 6 can be used. Here, x represents a support vector, and γ represents a kernel parameter for adjusting 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, as a method for determining or evaluating whether a specimen derived from a subject contains and / or does not contain the expression of a target gene derived from biliary tract cancer, or comparing and evaluating its expression level with a control derived from a healthy body, methods such as neural networks, k-nearest neighbor method, decision tree, and logistic regression analysis can be selected.

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

[0617] (a) a step of measuring the expression level of a target gene in a known sample of a tissue containing a biliary tract cancer-derived gene from a biliary tract cancer patient and / or a tissue not containing a biliary tract cancer-derived gene from a healthy person using the detection polynucleotide, kit or device (e.g., DNA chip) of the present invention,

[0618] (b) a step of generating discriminants of the above-mentioned formulas 1 to 3, 5 and 6 based on the expression level measured by (a),

[0619] (c) A step of measuring the expression level of the target gene in a specimen derived from a subject using the detection polynucleotide, kit or device (e.g., a DNA chip) of the present invention, substituting the measured value into the discriminant formula prepared in (b), and determining or evaluating whether the specimen contains and / or does not contain the target gene derived from biliary tract cancer based on the obtained result, or comparing and evaluating the expression level with a control derived from a healthy body.

[0620] Here, x in Formulas 1 to 3, 5 and 6 is an explanatory variable, which includes a value obtained by measuring a polynucleotide or a fragment thereof selected from the polynucleotide class described in Section 2 above. Specifically, the explanatory variable used to distinguish between biliary tract cancer patients and healthy bodies of the present invention is a gene expression level selected from, for example, the following (1) to (2).

[0621] (1) The gene expression level in the serum of a biliary tract cancer patient or a healthy person measured by any one of DNAs containing 15 or more consecutive bases of the base sequence represented by any one of SEQ ID NOs. 1 to 125 and 466 to 478 or its complementary sequence.

[0622] (2) The gene expression level in the serum of a biliary tract cancer patient or a healthy person measured by any one of DNAs containing 15 or more consecutive bases of the base sequence represented by any one of sequence numbers 126 to 148 or its complementary sequence.

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

[0624] Furthermore, the genes used as explanatory variables of the discriminant are preferably determined as follows. First, the total gene expression levels of the biliary tract cancer patient group and the healthy control group, which serve as the learning sample groups, are used as data sets. The difference in the expression levels of each gene between the two groups is determined using the P value of the t-test as a parametric analysis, the P value of the Mann-Whitney U test as a non-parametric analysis, or the P value of the Wilcoxon test.

[0625] When the risk (significance level) of the P value obtained by the test is, for example, less than 5%, 1% or 0.01%, it can be considered statistically significant.

[0626] To correct for the increased probability of a Type I error due to repeated testing, known methods such as the Bonferroni and Holm methods can be used (e.g., Yasushi Nagata et al., "The Fundamentals of Statistical Multiple Comparisons," Scientist (2007)). For example, the Bonferroni correction multiplies the P value obtained by the test by the number of test repetitions, i.e., the number of genes used in the analysis, and compares it with the desired significance level, thereby suppressing the probability of a Type I error in the overall test.

[0627] In addition, rather than testing, the absolute value of the expression ratio (fold change) of the median expression levels of the respective gene expression levels between the gene expression levels of the biliary tract cancer patient group and the gene expression levels of the healthy control group can be calculated to select genes used as the explanatory variable of the discriminant. In addition, the gene expression levels of the biliary tract cancer patient group and the healthy control group can be used to create an ROC curve, and the AUROC value can be used as a benchmark to select genes used as the explanatory variable of the discriminant.

[0628] Next, using the gene expression amount of the gene expression amount obtained here, the discriminant formula that can be calculated by the above-mentioned various methods is made. As a method for constructing a discriminant formula for obtaining the maximum discrimination accuracy, for example, there is the following method: a method for constructing a discriminant formula by all combinations of genes that meet the significance level of the P value; while the genes used to make the discriminant formula are increased one by one in the order of the difference in gene expression amount, and the method of repeatedly evaluating the same is repeated (Furey TS. et al., 2000, Bioinformatics., Vol. 16, p906-14). For this discriminant formula, the gene expression amount of other independent bile duct cancer patients or healthy bodies is substituted into the explanatory variable, and the discrimination result of the group to which the independent bile duct cancer patient or healthy body belongs is calculated. That is, by evaluating the diagnostic gene set (gene set) found with an independent specimen group and the discriminant formula constructed using the diagnostic gene set, a more general diagnostic gene set that can detect bile duct cancer and a method for discriminating bile duct cancer can be found.

[0629] Furthermore, the discriminant performance (generalizability) of this discriminant is preferably evaluated using the split-sample method. Specifically, the dataset is divided into a training sample group and a test sample group. Within the training sample group, statistical tests are performed to select genes and generate a discriminant. The discriminant performance is evaluated by calculating the accuracy, sensitivity, and specificity of the discriminant using the discriminant and the true group to which the test sample group belongs. Alternatively, the dataset can be left unsplit, with all samples subjected to statistical tests to select genes and generate a discriminant. New samples are then discriminated using the discriminant to calculate the accuracy, sensitivity, and specificity, thereby evaluating the discriminant performance.

[0630] The present invention provides polynucleotides for detection or disease diagnosis useful in the diagnosis and treatment of biliary tract cancer, methods for detecting biliary tract cancer using the polynucleotides, and kits and devices for detecting biliary tract cancer containing the polynucleotides. In particular, in order to select diagnostic genes and generate discriminants that exhibit accuracy exceeding that of existing biliary tract cancer diagnostic methods using the tumor markers CEA and CA19-9, the methods of the present invention compare, for example, the genes expressed in serum from patients whose biliary tract cancer was confirmed by detailed examinations such as computed tomography using contrast agents, with the genes expressed in serum from patients whose biliary tract cancer was not present, thereby constructing a diagnostic gene set and discriminant that exhibit accuracy exceeding that of CEA and CA19-9.

[0631] For example, any combination of one or more polynucleotides based on the base sequence shown in any one of sequence numbers 1 to 125, 466 to 478 or its complementary sequence as described above, and one or more polynucleotides based on the base sequence shown in any one of sequence numbers 126 to 148 or its complementary sequence, as appropriate, is set as a diagnostic gene set. Furthermore, a discriminant is constructed using the expression level of the diagnostic gene set in a specimen derived from a patient with grade I biliary tract cancer and a specimen derived from a grade II healthy body as determined by tissue diagnosis. As a result, by measuring the expression level of the diagnostic gene set in an unknown specimen, it is possible to distinguish with an accuracy of up to 100% whether the unknown specimen contains biliary tract cancer-derived genes or does not contain biliary tract cancer-derived genes.

[0632] Example

[0633] The present invention is further described in detail by the following examples, but the scope of the present invention is not limited by these examples.

[0634] [Reference Example 1]

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

[0636] Serum was collected from 100 healthy subjects who gave informed consent and 67 patients with biliary tract cancer who had no confirmed primary cancer other than the biliary tract (1 case at stage IA, 8 cases at stage IB, 8 cases at stage II, 3 cases at stage IIA, 5 cases at stage IIB, 14 cases at stage III, 2 cases at stage IIIB, 1 case at stage IVa, and 25 cases at stage IVb) using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.) to serve as a learning specimen group. Similarly, serum was collected from 50 healthy subjects who gave informed consent and 33 patients with biliary tract cancer in whom no primary cancer other than the biliary tract was confirmed (1 case at stage 0, 2 cases at stage I, 1 case at stage IA, 2 cases at stage IB, 2 cases at stage II, 5 cases at stage IIA, 4 cases at stage IIB, 5 cases at stage III, 1 case at stage IV, 1 case at stage IVa, and 9 cases at stage IVb) using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.) to serve as the test specimen group.

[0637] <Total RNA extraction>

[0638] Total RNA was obtained from 300 μL of serum obtained from a total of 250 people, including 150 healthy subjects and 100 biliary tract cancer patients, by combining the above-mentioned learning specimen group and test specimen group as specimens, using the RNA extraction reagent in the 3D-Gene (registered trademark) RNA extraction reagent from liquid sample kit (Toray Industries, Inc.) according to the manufacturer's protocol.

[0639] <Measurement of gene expression level>

[0640] For the total RNA obtained from the serum of 250 people (150 healthy people and 100 patients with biliary tract cancer) by combining the above-mentioned learning specimen group and test specimen group as specimens, the miRNA was fluorescently labeled using the 3D-Gene (registered trademark) miRNA Labeling kit (Toray Industries, Inc.) based on the instruction manual (ver2.20) formulated by the company. As an oligo DNA chip, a 3D-Gene (registered trademark) Human miRNA Oligo chip (Toray Industries, Inc.) equipped with probes having sequences complementary to 2,555 miRNAs in the miRNAs logged in miRBase version 20 was used. Hybridization of the miRNA in the total RNA with the probes on the DNA chip and post-hybridization washing were performed under strict conditions based on the instruction manual formulated by the company. A 3D-Gene (registered trademark) scanner (Toray Industries, Inc.) was used to scan the DNA chip, obtain an image, and digitize the fluorescence intensity using 3D-Gene (registered trademark) Extraction (Toray Industries, Inc.). The digitized fluorescence intensity was converted to a base-2 logarithmic value to represent the gene expression level. Blank values were subtracted, and missing values were replaced with a value obtained by subtracting 0.1 from the logarithmic value of the minimum gene expression level in each DNA chip. The results obtained were the gene expression levels of all miRNAs in the serum of 100 biliary tract cancer patients and 150 healthy subjects. Calculation and statistical analysis using the digitized 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] Serum was collected using Venoject II vacuum blood collection tubes VP-AS109K60 (Terumo Co., Ltd.) from 35 patients with colorectal cancer who had no confirmed cancer in other organs, 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. The samples were combined with 67 patients with biliary tract cancer in Reference Example 1 (1 patient at stage 0, 2 patients at stage I, 1 patient at stage IA, 4 patients at stage IB, 8 patients at stage II, 4 patients at stage IIA, 6 patients at stage IIB, 14 patients at stage III, 1 patient at stage IIIB, 25 patients at stage IV, and 1 patient at stage IVa) and 93 healthy subjects to form a learning specimen group. Similarly, serum was 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, all of whom had obtained informed consent and had no confirmed cancer in other organs, using Venoject II Vacuum Blood Collection Tubes VP-AS109K60 (Terumo Co., Ltd.). This serum was combined with 33 patients with biliary tract cancer from Reference Example 1 (1 patient at stage IA, 6 patients at stage IB, 2 patients at stage II, 4 patients at stage IIA, 3 patients at stage IIB, 5 patients at stage III, 1 patient at stage IIIB, and 11 patients at stage IV) and 57 healthy subjects to form a test sample group. The following total RNA extraction and gene expression level measurement and analysis were performed in the same manner as in Reference Example 1.

[0644] [Example 1]

[0645] <Selection of Genetic Markers Using Samples from a Training Sample Group and Evaluation Method for Biliary Tract Cancer Discrimination Performance of Individual Genetic Markers Using Samples from a Test Sample Group>

[0646] This embodiment studies a method of selecting gene markers for distinguishing biliary tract cancer from healthy subjects from a learning specimen group, and evaluating the biliary tract cancer discrimination performance of the selected gene markers separately in specimens of a test specimen group independent of the learning specimen group.

[0647] Specifically, first, the miRNA expression levels of the learning sample group and the test sample group obtained in Reference Example 1 were combined and normalized using quantile normalization.

[0648] Next, the diagnostic genes were selected using the learning sample group. Here, in order to obtain more reliable diagnostic markers, only genes with a gene expression level of 2 to the power of 6 or more in more than 50% of the samples in either the biliary tract cancer patient group or the healthy body group of the learning sample group were selected. Furthermore, as statistically significant genes for discriminating between the biliary tract cancer patient group and the healthy body group, the P value obtained by the two-sided t-test assuming equal variance was Bonferroni-corrected for each gene expression level, and genes satisfying p < 0.01 were obtained as gene markers used as explanatory variables of the discriminant, which are recorded 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 are markers for cholangiocarcinoma relative to healthy subjects.

[0650] Furthermore, using the expression levels of these genes as indicators, Fisher's discriminant analysis was used to generate a discriminant for the presence of biliary tract cancer. Specifically, a discriminant was generated by inputting a newly discovered polynucleotide consisting of a base sequence represented by any of SEQ ID NOs. 1 to 125 from among the 125 genes selected in the learning sample set into Equation 2. The calculated accuracy, sensitivity, and specificity are shown in Table 3. The discriminant coefficient and constant term are also shown in Table 4.

[0651] Using the discriminant formula prepared above, the accuracy, sensitivity, and specificity in the test specimen group were calculated, and the discrimination performance of the selected polynucleotide was verified using an independent specimen (Table 3). For example, when the expression level measurement value of the base sequence shown in sequence number 1 was compared between the healthy subjects (100 people) and the biliary tract cancer patients (67 people) in the learning specimen group, it was shown that the gene expression level measurement value of the biliary tract cancer patient group was significantly lower than that of the healthy subject group (refer to Figure 2 Left), further, the results can be reproduced in the healthy subjects (50 people) and biliary tract cancer patients (33 people) in the test sample group (refer to Figure 2 Right). Similarly, for the other polynucleotides shown in sequence numbers 2 to 125, results were obtained showing that the gene expression levels of the biliary tract cancer patient group were significantly lower (-) or higher (+) than those of the healthy group (Table 2), and these results were verified in the test specimen group. In addition, for example, with respect to the base sequence shown in sequence number 1, the hit rate of biliary tract cancer detection was calculated using the threshold value (5.69) for distinguishing the two groups set in the learning specimen group. The results showed 33 true positives, 49 true negatives, 1 false positive, and 0 false negatives. Using these values as the detection performance, an accuracy of 99%, a sensitivity of 100%, and a specificity of 98% were obtained. The detection performance of all the polynucleotides shown in sequence numbers 1 to 125 was calculated in this way and is recorded in Table 3.

[0652] Among the polynucleotides consisting of the base sequences represented by SEQ ID NOs. 1 to 125 shown in Table 2, for example, SEQ ID NOs. 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 consisting 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 75.9%, respectively, in the test sample group. .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, according to the comparative example described later, the sensitivity of the existing markers CEA in the test sample group is 33.3%, and the sensitivity of CA19-9 is 59.4% (Table 5). This proves that in the test sample group, for example, the markers represented by 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, 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 alone diagnose biliary tract cancer with a higher sensitivity than the existing blood tumor marker CA19-9.

[0653] Furthermore, for example, nine polynucleotides consisting of the base sequences represented by SEQ ID NOs: 1, 2, 3, 4, 10, 11, 12, 23, and 64 correctly identified all six stage 0 and stage 1 (including IA and IB) biliary tract cancer specimens in the test sample group 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 identify biliary tract cancer in all test specimens, including those in the extrahepatic bile duct, intrahepatic bile duct, gallbladder, and papillary regions of the bile duct. In particular, they can detect cancer in the lower bile duct, papillary region, and intrahepatic bile duct, where the prognosis is considered poor, and in which the disease tends to progress asymptomatically.

[0655] [Example 2]

[0656] <Method for evaluating biliary tract cancer discrimination performance using a combination of multiple gene markers from a test sample group>

[0657] This example studies a method for evaluating the performance of biliary tract cancer discrimination using a combination of gene markers selected in Example 1. Specifically, Fisher's discriminant analysis was performed on 7,750 combinations of any two of the expression level measurement values of the polynucleotides consisting of the base sequences shown in sequence numbers 1 to 125 selected in Example 1, and a discriminant formula for discriminating the presence or absence of biliary tract cancer was constructed. Next, the accuracy, sensitivity, and specificity in the test specimen group were calculated using the discriminant formula prepared above, and the discrimination performance of the selected polynucleotides was verified using an independent specimen. The result of implementing biliary tract cancer discrimination in the test specimen group using a combination of the above-mentioned 7,750 types of polynucleotide expression level measurement values was that, for example, when the expression level measurement values of the polynucleotides consisting of the base sequences shown in sequence numbers 2 and 4 were compared between healthy subjects (50 people) and biliary tract cancer patients (33 people) in the test specimens, a scatter plot showing a significant separation of the expression level measurement values of the healthy group and the biliary tract cancer patient group was obtained in the learning specimen group (refer to Figure 3 Left), and the results were also reproduced in the test sample group (refer to Figure 3Right). Similarly, even in any other combination of the expression level measurement values of the newly discovered polynucleotides consisting of the base sequences shown in sequence numbers 1 to 125, a scatter plot that significantly separates the expression level measurement values of the healthy group and the biliary tract cancer patient group was obtained, and these results were verified in the test specimen group. In addition, for example, with respect to the base sequences shown in sequence numbers 2 and 4, the function (0=5.16x+y+48.11) for discriminating between the two groups set in the learning specimen group was used to calculate the hit rate of biliary tract cancer detection, and the results were 33 true positives, 48 true negatives, 2 false positives, and 0 false negatives. From these values, an accuracy of 98%, a sensitivity of 100%, and a specificity of 96% were obtained as the detection performance. In this way, the detection performance of all combinations of any two of the expression level measurement values of the newly discovered polynucleotides consisting of the base sequences shown in sequence numbers 1 to 125 was calculated. As an example, 124 combinations of polynucleotides consisting of the base sequence represented by SEQ ID NO: 1 and polynucleotides consisting of base sequences represented by other SEQ ID NOs, and their detection performance, are listed in Table 6. For example, combinations of expression level measurement values for polynucleotides consisting of the base sequences represented by SEQ ID NOs: 1 and 7, 1 and 9, 1 and 25, and 1 and 66 all showed a sensitivity of 100% in the test sample group. Thus, 6,316 combinations of polynucleotide expression level measurement values exceeding the sensitivity of CA19-9, an existing marker (75.8% as shown in Table 5), were obtained in the test sample group. In these combinations, all of the 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 combination of two of the expression level measurement values for polynucleotides consisting of the base sequences represented by SEQ ID NOs: 1 to 125 distinguishes biliary tract cancer with a sensitivity higher than that of CA19-9.

[0658] Furthermore, among the 7,750 combinations of any two expression level measurements of polynucleotides consisting of the base sequences shown in SEQ ID NOs: 1 to 125, there were 1,290 combinations of two that correctly identified all six stage 0 and stage 1 (including stage IA and IB) biliary tract cancer specimens included in the test specimen group as biliary tract cancer. In these 1,290 combinations of two, the polynucleotides consisting of the base sequences shown in SEQ ID NOs: 1 to 125 were used at least once. In other words, these polynucleotides can also detect early biliary tract cancer and contribute to the early diagnosis of biliary tract cancer.

[0659] Thus, even if 3, 4, 5, 6, 7, 8, 9, 10, or more expression level values of polynucleotides consisting of the base sequences shown in SEQ ID NOs: 1 to 125 are combined, a marker for detecting biliary tract cancer with excellent sensitivity can be obtained. For example, for the polynucleotides consisting of the base sequences shown in SEQ ID NOs: 1 to 125 selected in Example 1, the detection performance was calculated using combinations of one or more miRNAs, starting from the top miRNA, in descending order of statistically significant P values. The results showed that the sensitivity in the test sample group was 100% for 1 miRNA, 100% for 2 miRNAs, 100% for 3 miRNAs, 100% for 5 miRNAs, 100% for 10 miRNAs, 100% for 20 miRNAs, 100% for 50 miRNAs, and 100% for 100 miRNAs. These sensitivities are higher than those of existing blood tumor markers, demonstrating that even combinations of multiple miRNAs can be used to create excellent markers for detecting biliary tract cancer. The combination of multiple miRNAs is not limited to combinations in the order of statistical significance as described above; any combination of multiple miRNAs can be used to detect biliary tract cancer.

[0660] From these results, it can be said that all polynucleotides consisting of the base sequences represented by SEQ ID NOs: 1 to 125 are excellent diagnostic markers 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] Study specimen group

[0678]

[0679]

[0680] [Table 5-2]

[0681] Test specimen group

[0682] 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 O 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.g 777 P91 IVb 2.3 4308 Sensitivity (%) 33.3 59.4

[0683] In Table 5, a CEA value of 5 ng / ml or less was designated as "-", a CA19-9 value of 37 U / ml or less was designated as "-", and values exceeding these values were designated as "+".

[0684] [Table 6]

[0685]

[0686]

[0687]

[0688]

[0689] [Example 3]

[0690] <Selection of Genetic Markers Using All Samples and Method for Evaluating the Biliary Tract Cancer Discrimination Performance of the Resultant Genetic Markers>

[0691] In this example, the samples of the learning sample group and the test sample group used in the above-mentioned Examples 1 and 2 were combined and all the samples were used to select gene markers and evaluate their biliary tract cancer discrimination performance.

[0692] Specifically, the miRNA expression levels in the sera of 100 biliary tract cancer patients and 150 healthy subjects obtained from the above-mentioned Reference Example 1 were normalized using quantile normalization. In order to obtain more reliable diagnostic markers, only genes with a gene expression level of 2 to the power of 6 or more in more than 50% of the specimens in either the biliary tract cancer patient group or the healthy body group were selected. Furthermore, in order to obtain statistical significance for discriminating between the biliary tract cancer patient group and the healthy body group, the P value obtained by the two-sided t-test assuming equal variance was Bonferroni-corrected for each gene expression level, and genes satisfying p < 0.01 were selected as the explanatory variables of the discriminant equation. The gene markers used are recorded in Table 7. Thus, in addition to the genes listed in Table 2, 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 were also found. -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 genes were used as biliary tract cancer markers relative to healthy subjects. Similar to the polynucleotides represented by sequence numbers 1 to 125, the polynucleotides represented by sequence numbers 126 to 148 also showed that the expression levels in the biliary tract cancer patient group were significantly lower (-) or higher (+) than those in the healthy group (Table 7), and these results were verified in the test specimen group. By using the expression level measurement values of the genes listed in Table 7 alone or in combination with the expression level measurement values of the genes listed in Table 2, newly obtained specimens can be identified by the methods described in Examples 1 and 2.

[0693] [Table 7]

[0694]

[0695]

[0696]

[0697]

[0698] [Example 4]

[0699] <Method for evaluating biliary tract cancer-specific discrimination performance using a combination of multiple gene markers using test sample group specimens>

[0700] In this example, using the learning sample group described in Reference Example 2 as the subject, the same method as described in Example 1 was used to compare the expression levels of miRNA genes in the serum of patients with biliary tract cancer with a control group consisting of healthy individuals, patients with colorectal cancer, patients with gastric cancer, patients with esophageal cancer, patients with liver cancer, and patients with benign pancreatic and biliary tract diseases, to select additional diagnostic gene markers. The results were evaluated using one or more markers selected from a group combining the selected additional diagnostic gene markers (SEQ ID NOs. 466 to 478; see Table 1) with the gene markers selected in Example 1 to evaluate the specific discriminatory performance for biliary tract cancer.

[0701] Specifically, first, the miRNA expression levels of the learning specimen group and the test specimen group obtained by the above-mentioned reference example 2 are combined and normalized by quantile normalization. Next, Fisher's discriminant analysis is performed on 1 to 4 combinations of expression level measurement values of any one of the polynucleotides consisting of at least one base sequence represented by sequence numbers 1 to 148, 466 to 478, to construct a discriminant formula for discriminating the presence or absence of biliary tract cancer. Next, the biliary tract cancer patient group is used as the positive specimen group, and the healthy group, colorectal cancer patient group, gastric cancer patient group, esophageal cancer patient group, liver cancer patient group and pancreatic biliary benign disease patient group are used as the negative specimen group. The accuracy, sensitivity and specificity in the test specimen group are calculated using the discriminant formula made above, and the discrimination performance of the selected polynucleotides is verified using independent specimens.

[0702] Most of the polynucleotides composed of the base sequences shown by the above sequence numbers (sequence numbers 1 to 148 and 466 to 478 corresponding to the miRNA markers in Table 1) or their complementary sequences can not only provide relatively high accuracy, sensitivity, and specificity in determining the presence or absence of biliary tract cancer, but can also specifically identify biliary tract cancer from other cancers. As the polynucleotides listed as being capable of specifically binding to a target marker, for example, a combination of multiple polynucleotides selected from the 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 (cancer type-specific polynucleotide group 1), a combination comprising at least one polynucleotide 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 (cancer type-specific polynucleotide group 2) can specifically identify biliary tract cancer from other cancers with high accuracy.

[0703] The number of combinations of the above-mentioned cancer type-specific polynucleotides can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10, and a discrimination accuracy of more than 80% can be shown in combinations of 4 or more.

[0704] Specifically, the discrimination accuracy when the polynucleotide consisting of the base sequence shown in sequence number 4 or its complementary sequence is measured is as follows. When the polynucleotide consisting of the base sequence shown in sequence number 4 or its complementary sequence is used alone (1) for measurement, the accuracy is 81.9% in the learning specimen group and 76.9% in the test specimen group (Table 8). In addition, for example, when the combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in sequence number 4 or its complementary sequence is used for measurement, the highest accuracy is 86.0% in the learning specimen group and 85.3% in the test specimen group (Table 9; "sequence number" in the table represents the combination of the sequence numbers of the two polynucleotides used). In addition, for example, when a combination of three polynucleotides including at least one polynucleotide consisting of the base sequence shown in sequence number 4 or its complementary sequence was used for measurement, the highest accuracy was 89.5% in the learning specimen group, and the accuracy was 90.4% in the test specimen group (Table 10; the "sequence number" in the table represents the combination of the sequence numbers of the three polynucleotides used). In addition, for example, when a combination of four polynucleotides including at least one polynucleotide consisting of the base sequence shown in sequence number 4 or its complementary sequence was used for measurement, the highest accuracy was 91.1% in the learning specimen group, and the accuracy was 92.3% in the test specimen group (Table 11; the "sequence number" in the table represents the combination of the sequence numbers of the four polynucleotides used).

[0705] Specifically, the discrimination accuracy when the polynucleotide consisting of the base sequence shown in sequence number 5 or its complementary sequence is measured is as follows. When the polynucleotide consisting of the base sequence shown in sequence number 5 or its complementary sequence is used alone (1) for measurement, the accuracy is 79.0% in the learning sample group and 80.8% in the test sample group (Table 8). In addition, for example, when the combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in sequence number 5 or its complementary sequence is used for measurement, the highest accuracy is 81.9% in the learning sample group and 86.5% in the test sample group (Table 9). In addition, for example, when the combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in sequence number 5 or its complementary sequence is used for measurement, the highest accuracy is 87.6% in the learning sample group and 89.7% in the test sample group (Table 10). In addition, for example, when measurements were performed using a combination of four polynucleotides each comprising at least one polynucleotide consisting of the base sequence shown in sequence number 5 or its complementary sequence, the highest accuracy was 93.0% in the learning sample group and 91.0% in the test sample group (Table 11).

[0706] Specifically, the discrimination accuracy when the polynucleotide consisting of the base sequence shown in sequence number 12 or its complementary sequence was measured is as follows. When the polynucleotide consisting of the base sequence shown in sequence number 12 or its complementary sequence was used alone (1) for measurement, the accuracy was 80.6% in the learning sample group and 76.9% in the test sample group (Table 8). In addition, for example, when the combination of two polynucleotides containing at least one polynucleotide consisting of the base sequence shown in sequence number 12 or its complementary sequence was used for measurement, the highest accuracy was 86.3% in the learning sample group and 85.9% in the test sample group (Table 9). In addition, for example, when the combination of three polynucleotides containing at least one polynucleotide consisting of the base sequence shown in sequence number 12 or its complementary sequence was used for measurement, the highest accuracy was 90.2% in the learning sample group and 91.7% in the test sample group (Table 10). In addition, for example, when measurements were performed using a combination of four polynucleotides each comprising at least one polynucleotide consisting of the base sequence shown in sequence number 12 or its complementary sequence, the highest accuracy was 93.0% in the learning sample group and 94.2% in the test sample group (Table 11).

[0707] Specifically, the discrimination accuracy when the polynucleotide consisting of the base sequence shown in sequence number 15 or its complementary sequence was measured is as follows. When the polynucleotide consisting of the base sequence sh...

Claims

1. A nucleic acid probe that can specifically bind to a polynucleotide of hsa-miR-665, a biliary tract cancer marker, and / or a primer that specifically recognizes and amplifies the polynucleotide, and a nucleic acid probe that can specifically bind to a polynucleotide of hsa-miR-125a-3p, a biliary tract cancer marker, and / or a primer that specifically recognizes and amplifies the polynucleotide, for producing a kit for detecting biliary tract cancer.

2. The use according to claim 1, wherein the nucleic acid probe that specifically binds to the hsa-miR-665 polynucleotide and / or the primer that specifically recognizes and amplifies the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a) to (d) below. (a) a polynucleotide consisting of the base sequence represented by SEQ ID NO: 66, or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, (b) a polynucleotide comprising the base sequence represented by SEQ ID NO: 66, (c) a polynucleotide consisting of a base sequence complementary to the base sequence represented by SEQ ID NO: 66 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, and (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by SEQ ID NO: 66 or a base sequence in which u is t in the base sequence, The nucleic acid probe capable of specifically binding to the hsa-miR-125a-3p polynucleotide and / or the primer that specifically recognizes and amplifies the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a') to (d') below, (a') a polynucleotide consisting of the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, (b') a polynucleotide comprising the base sequence shown in SEQ ID NO: 1, (c') a polynucleotide consisting of a base sequence complementary to the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, and (d') A polynucleotide comprising a base sequence complementary to the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence.

3. Use of a nucleic acid probe that can specifically bind to a polynucleotide of hsa-miR-665, a marker for biliary tract cancer, and / or a primer that specifically identifies and amplifies the polynucleotide, and a nucleic acid probe that can specifically bind to a polynucleotide of hsa-miR-125a-3p, a marker for biliary tract cancer, and / or a primer that specifically identifies and amplifies the polynucleotide, for manufacturing a device for detecting biliary tract cancer.

4. The use according to claim 3, wherein the nucleic acid probe capable of specifically binding to the hsa-miR-665 polynucleotide and / or the primer that specifically recognizes and amplifies the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a) to (d) below. (a) a polynucleotide consisting of the base sequence represented by SEQ ID NO: 66, or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, (b) a polynucleotide comprising the base sequence represented by SEQ ID NO: 66, (c) a polynucleotide consisting of a base sequence complementary to the base sequence represented by SEQ ID NO: 66 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, and (d) a polynucleotide comprising a base sequence complementary to the base sequence represented by SEQ ID NO: 66 or a base sequence in which u is t in the base sequence, The nucleic acid probe capable of specifically binding to the hsa-miR-125a-3p polynucleotide and / or the primer that specifically recognizes and amplifies the polynucleotide is a polynucleotide selected from the polynucleotides shown in (a') to (d') below, (a') a polynucleotide consisting of the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, (b') a polynucleotide comprising the base sequence shown in SEQ ID NO: 1, (c') a polynucleotide consisting of a base sequence complementary to the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence, or a fragment of the polynucleotide comprising 15 or more consecutive bases, and (d') A polynucleotide comprising a base sequence complementary to the base sequence shown in SEQ ID NO: 1 or a base sequence in which u is t in the base sequence.

5. The use according to claim 3 or 4, wherein the device is a device for performing assays by hybridization techniques. The method according to claim 5 , wherein the hybridization technique is nucleic acid array technique.

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