Biomarker LINC01549 related to hepatocellular carcinoma and application thereof

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

Active Publication Date: 2018-10-16
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.

Method used

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  • Biomarker LINC01549 related to hepatocellular carcinoma and application thereof
  • Biomarker LINC01549 related to hepatocellular carcinoma and application thereof
  • Biomarker LINC01549 related to hepatocellular carcinoma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Example 1 Screening of gene markers related to hepatocellular carcinoma

[0087] 1. Sample collection

[0088] Collect 50 HBV-infected hepatocellular carcinoma patients’ cancer tissues and corresponding para-cancerous tissue samples, randomly select 8 cases for high-throughput sequencing, and all patients have informed consent. All the above-mentioned specimens were obtained through the organization ethics committee agree.

[0089] 2. Preparation of RNA samples

[0090] Use QIAGEN's tissue RNA extraction kit to extract tissue RNA, and follow the specific steps in the instructions.

[0091] 3. Quality analysis of RNA samples

[0092] The RNA extracted above was subjected to agarose gel electrophoresis, the concentration and purity of the extracted RNA were detected by Nanodrop2000, the integrity of RNA was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent 2100. A single library construction requires a total amount of RNA of 5ug, a concentration ...

Embodiment 2

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

[0104] 1. Use the previously collected 50 patients with cancer tissue samples and adjacent tissue samples to verify the differential expression of the LINC01549 gene with a large sample of QPCR.

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

[0106] 3. Reverse transcription:

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

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

[0109] 3) Combine the reaction mixture with 4μl 5X buffer, 1μl dNTP (10mM), 0.5μl M-MLV reverse transcriptase, 0.5μl ribonuclease (RNase) inhibitor, 10μl polyA reaction mixture and 4μl ribonuclease-free water (RNase freewater) mix...

Embodiment 3

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

[0120] 1. Cell culture

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

[0122] 2. RNA extraction

[0123] Use QIAGEN's Cell RNA Extraction Kit to extract total cell RNA. Refer to the instructions for specific operations.

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

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

[0126] 5. Results

[0127] The result is figure 2 As shown, compared with normal liver cell lines, the expression of LINC01549 gene was up-regulated in hepatocellular carcinoma cells HepG2 and Huh7, and the ...

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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 its application. Background technique [0002] Liver cancer is a common malignant disease with a high fatality rate. The incidence of liver cancer ranks sixth among all cancers worldwide, and its fatality rate ranks second (Torre LA, Bray F, Siegel RL, Ferlay J, Lortet -Tieulent J, Jemal A. Global cancer statistics, 2012. CA: a cancer journal forclinicians. 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 of liver cancer and the application of targeted drugs such as sorafenib have significantly improved the quality of life of patients with liver cancer. However, since most patients are already in the advanced stage at the time of diagnosis and the hetero...

Claims

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Application Information

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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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