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A biomarker linc01549 related to hepatocellular carcinoma and its application

A kind of hepatocellular carcinoma, substance technology, applied in the field of biomedicine

Active Publication Date: 2020-09-22
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of effective biomarkers for the clinical diagnosis and treatment of hepatocellular carcinoma. Therefore, it is of great significance to find lncRNAs related to the occurrence and development of hepatocellular carcinoma.

Method used

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  • A biomarker linc01549 related to hepatocellular carcinoma and its application
  • A biomarker linc01549 related to hepatocellular carcinoma and its application
  • A biomarker linc01549 related to hepatocellular carcinoma and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Screening for Gene Markers Related to Hepatocellular Carcinoma

[0087] 1. Sample collection

[0088] The cancer tissues and corresponding paracancerous tissue samples of 50 HBV-infected patients with hepatocellular carcinoma were collected, and 8 cases were randomly selected for high-throughput sequencing. The patients all gave informed consent. agree.

[0089] 2. Preparation of RNA samples

[0090] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0091] 3. Quality analysis of RNA samples

[0092] The above extracted RNA was subjected to agarose gel electrophoresis, the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of the RNA was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. The total amount of RNA required for a single library construction is 5ug, the concentration is ≥20...

Embodiment 2

[0103] Example 2 QPCR sequencing to verify the differential expression of the LINC01549 gene

[0104] 1. Using the previously collected cancer tissue samples and paracancerous tissue samples from 50 patients, the differential expression of the LINC01549 gene was verified by large sample QPCR.

[0105] 2. The RNA extraction steps are the same as in Example 1.

[0106] 3. Reverse transcription:

[0107] 1) Mix 1 μg of total RNA template with 2 μl 10× buffer, 2 μl dATP (10 mM), 0.5 μl polyA polymerase, 0.5 μl ribonuclease (RNase) inhibitor and ribonuclease-free water (RNase free water), volume Finally, 20 μl was incubated at 37°C for 1 hour;

[0108] 2) Add 1 μl 0.5 μg / μl Oligo(dT)-specific RT primer to the reaction tube, incubate at 70°C for 5 minutes, and immediately place it on ice for at least 2 minutes;

[0109] 3) Mix the reaction mixture with 4 μl 5× buffer, 1 μl dNTP (10 mM), 0.5 μl M-MLV reverse transcriptase, 0.5 μl ribonuclease (RNase) inhibitor, 10 μl polyA reactio...

Embodiment 3

[0119] Example 3 Differential expression of LINC01549 gene in hepatocellular carcinoma cell lines

[0120] 1. Cell culture

[0121] Human hepatocellular carcinoma cell lines HepG2, Huh7 and normal liver cell line HL-7702 were incubated in DMEM containing 10% fetal bovine serum and 1% P / S at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0122] 2. Extraction of RNA

[0123] Total cellular RNA was extracted using QIAGEN's cellular RNA extraction kit, and the specific operation was referred to the instructions.

[0124] 3. The specific steps of reverse transcription are the same as in Example 2

[0125] 4. The specific steps of QPCR amplification inspection are the same as in Example 2

[0126] 5. Results

[0127] The result is as figure 2 As shown, compared with normal liver cell lines, the expression of LINC01549 gene was up-r...

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Abstract

The invention discloses a biomarker LINC01549 related to hepatocellular carcinoma and application thereof. The invention finds that expression of the LINC01549 is up-regulated in a patient suffering from the hepatocellular carcinoma for the first time, and after further verification through a large sample, a result shows that the LINC01549 as a detection substance has a relatively high positive detection rate; experiments further prove that the LINC01549 is related to proliferation, migration and invasion of hepatoma carcinoma cells, which indicates that the LINC01549 can be used as a medicinetarget for clinical treatment of the hepatocellular carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a biomarker LINC01549 related to hepatocellular carcinoma and an application thereof. Background technique [0002] Liver cancer is a common malignant tumor disease with a high mortality rate. The incidence of liver cancer ranks sixth among all cancers worldwide, and its lethality ranks second (Torre LA, Bray F, Siegel RL, Ferlay J, Lortet -Tieulent J, Jemal A. Global cancer statistics, 2012. CA: a cancer journal for clinicians. 2015; 65:87-108.). my country is a big country with liver cancer, with hundreds of thousands of new patients each year, accounting for more than half of the world's liver cancer patients. The current advanced surgical treatment for liver cancer and the application of targeted drugs such as sorafenib have significantly improved the quality of life of patients with liver cancer. Poor treatment effect and poor prognosis (McGlynn KA, Petrick JL, London WT. Globalep...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K31/7105A61K48/00A61P35/00A61P35/04
CPCA61K31/7105A61P35/00A61P35/04C12Q1/6886C12Q2600/106C12Q2600/178
Inventor 董东石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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