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Seven serum exosome micro-ribonucleic acids (miRNAs) related to diagnosis and treatment of liver cancer, and applications of seven serum exosome miRNAs

A technology for exosomes and liver cancer, applied in the direction of DNA/RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve the problems of insufficient sensitivity and specificity to meet the early detection of liver cancer and poor accuracy

Inactive Publication Date: 2018-06-01
SUZHOU GENEPHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main blood marker used for the diagnosis of liver cancer is alpha-fetoprotein (AFP), but its accuracy is not good, and its sensitivity and specificity are not enough to meet the early detection of liver cancer

Method used

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  • Seven serum exosome micro-ribonucleic acids (miRNAs) related to diagnosis and treatment of liver cancer, and applications of seven serum exosome miRNAs
  • Seven serum exosome micro-ribonucleic acids (miRNAs) related to diagnosis and treatment of liver cancer, and applications of seven serum exosome miRNAs
  • Seven serum exosome micro-ribonucleic acids (miRNAs) related to diagnosis and treatment of liver cancer, and applications of seven serum exosome miRNAs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0137] Example 1. Discovery and verification of seven serum exosomal miRNAs as liver cancer markers

[0138] There were 110 serum samples, 80 of which were from 80 patients with liver cancer diagnosed in the First Affiliated Hospital of Soochow University from December 2015 to January 2017 (volunteers with informed consent, diagnosed as stage I-IV liver cancer by histopathology Patients, without blood collection before surgery and radiotherapy and chemotherapy after admission), and the remaining 30 were from 30 healthy people (volunteers with informed consent) who underwent disease screening at the same time. The information of 80 patients and 30 healthy people is shown in Table 1.

[0139] Table 1

[0140]

[0141] Each serum sample was tested separately.

[0142] 1. Extraction and identification of serum exosomes

[0143] 1. Take serum samples and extract exosomes by PEG-base precipitation method.

[0144] 2. Take the exosomes obtained in step 1 and observe them with ...

Embodiment 2

[0175] Embodiment 2, cell proliferation test

[0176] The test substance is: hsa-miR-106a mimic or hsa-miR-106a inhibitor.

[0177] The hsa-miR-106a mimic is a double-stranded RNA formed by the single-stranded RNA shown in sequence 29 of the sequence listing and the single-stranded RNA shown in sequence 30 of the sequence listing.

[0178] Sequence 29 of the sequence listing: 5'-AAAAGUGCUUACAGUGCAGGUAG-3';

[0179] Sequence 30 of the Sequence Listing: 5'-ACCUGCACUGUAAGCACUUUUUU-3'.

[0180] The hsa-miR-106a inhibitor is a single-stranded RNA shown in sequence 31 of the sequence listing.

[0181] Sequence 31 of the Sequence Listing: 5'-CUACCUGCACUGUAAGCACUUUU-3'.

[0182] The cells to be tested are: HEPG2 cells or SMMC-7721 cells.

[0183] 1. Use DMEM culture medium to suspend the cells to be tested, and then inoculate the cell suspension into a 96-well plate, 100 μl per well (containing 10 5 cells), and then cultured statically at 37°C for 24 hours.

[0184] 2. Preparati...

Embodiment 3

[0190] Embodiment 3, cell invasion test

[0191] Test object is with embodiment 2.

[0192] The cells to be tested are the same as in Example 2.

[0193]1. Use DMEM culture medium to suspend the cells to be tested, and then inoculate the cell suspension into a 96-well plate, 100 μl per well (containing 10 5 cells), and then cultured statically at 37°C for 24 hours.

[0194] 2. Preparation method of each mixture: 20 μl of healthy exosomes (containing 10 μg of exosomes, calculated as protein) prepared in step 1 of Example 1 were mixed with 200 pmol of the test substance to obtain a mixture.

[0195] 3. After completing step 1, take the 96-well plate, discard the supernatant, add 1 part of the mixture prepared in step 2 to each well, then culture at 37°C for 24 hours, and collect the cells. Set up exosomes only, instead of adding the control to the mixture.

[0196] 4. Matrigel was melted at 4°C, and 100 μl of the melted matrigel was mixed with 400 μl of serum-free DMEM cultu...

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Abstract

The invention discloses seven serum exosome micro-ribonucleic acids (miRNAs) related to diagnosis and treatment of liver cancer, and applications of the seven serum exosome miRNAs. The invention firstly discovers that hsa-miR-122, hsa-miR-125b, hsa-miR-145, hsa-miR-192, hsa-miR-194, hsa-miR-29a and hsa-miR-106a in serum exosomes can be used as biomarkers of the liver cancer, have a higher diagnostic value, provide a rapid and accurate diagnosis way for clinical use, and enable the diagnosis of the liver cancer to be more convenient and easier to implement. The invention also further discoversmatters for inhibiting specific miRNA, and the matters can inhibit the proliferation / growth of cells with the liver cancer, so that a foundation is laid for the clinic treatment of the liver cancer.

Description

technical field [0001] The invention belongs to the field of biotechnology and diagnosis, and specifically relates to seven serum exosomal miRNAs related to the diagnosis and treatment of liver cancer and their applications. The seven serum exosomal miRNAs are specifically hsa-miR-122, hsa-miR-125b, hsa- miR-145, hsa-miR-192, hsa-miR-194, hsa-miR-29a and hsa-miR-106a. Background technique [0002] Liver cancer is a malignant tumor with the fourth highest incidence rate and the second highest mortality rate in the world. Because of the relationship between diet, alcoholic liver disease, especially liver cirrhosis, and hepatitis B, liver cancer has a high incidence and high risk in China. Many liver cancers are usually found in clinically advanced stages, making the 5-year survival rate of liver cancer in my country less than 10%. Therefore, the early diagnosis of liver cirrhosis and liver cancer is very important. At present, the main blood marker used for the diagnosis of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K31/7105A61P35/00C12N15/11
CPCA61K31/7105C12Q1/6886C12Q2600/178
Inventor 胡荣宽赵毓斌王晓娜薛小峰张佩琢
Owner SUZHOU GENEPHARMA
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