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Reagents for detecting and targeting lncRNA biomarkers and their application in hepatocellular carcinoma

A technology of hepatocellular carcinoma and reagents, applied in the direction of microbial determination/testing, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem of unsatisfactory long-term prognosis

Active Publication Date: 2021-03-23
SICHUAN PROVINCIAL PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with the development of effective surgical techniques and improvements in diagnosis, the prognosis of patients with primary liver cancer has improved compared to the past. However, the recurrence rate of patients with hepatocellular carcinoma after surgical resection is as high as 50-70%. , the long-term prognosis of patients remains unfavorable

Method used

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  • Reagents for detecting and targeting lncRNA biomarkers and their application in hepatocellular carcinoma
  • Reagents for detecting and targeting lncRNA biomarkers and their application in hepatocellular carcinoma
  • Reagents for detecting and targeting lncRNA biomarkers and their application in hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 QPCR sequencing verification of the expression of the CTD-2561J22.5 gene

[0056] 1. Sample collection

[0057] A total of 42 cases of liver cancer tissues and corresponding normal paracancerous tissues were selected from patients diagnosed with liver cancer. All patients who were enrolled in the group had not received chemotherapy or radiotherapy before surgery.

[0058] 2. Trizol method to extract RNA from samples

[0059] Total RNA was extracted from tissue samples using Trizol reagent. Cut the tissue with scissors, add 1ml pre-cooled TRizol, and mix until a transparent solution. Transfer the lysate to a 1.5ml centrifuge tube and place it at room temperature for 5 minutes to completely separate the nucleic acid-protein complex. Chloroform was added, shaken, and centrifuged. Transfer the colorless supernatant to another centrifuge tube, add isopropanol to the supernatant, let it stand, centrifuge, remove the supernatant, add ethanol, centrifuge, remove t...

Embodiment 2

[0073] Example 2 Silencing of CTD-2561J22.5 gene and its influence on cells

[0074] 1. Cell culture

[0075] The liver cancer cell line HepG2 and the normal cell line LO2 were purchased from ATCC. All cell lines were cultured in DMEM containing 10% fetal bovine serum and 1% P / S at 37°C, 5% CO 2 cultured in an incubator. The cell growth was observed every day, and the medium was changed every other day.

[0076] 2. Transfection

[0077] The siRNA against CTD-2561J22.5 was designed and synthesized by Shanghai Jima Pharmaceutical Technology Co., Ltd., the specific sequence is 5'-AACAUGUUUUCUUCUCAUGAU-3', SEQ ID NO.5; the antisense strand is 5'-CAUGAGAAGAAAACAUGUUUA-3', SEQ ID NO.6, the control is general siRNA-NC.

[0078] According to the instructions of Lipofectamine 3000 reagent, mix the liposome and OPTI-MEM reduced serum medium, siRNA and OPTI-MEM medium separately, leave at room temperature for 5min, then mix the liposome, siRNA and OPTI-MEM medium, Leave it for 20min....

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Abstract

The invention discloses a reagent for detecting and targeting a lncRNA biomarker and application of the reagent to hepatocellular carcinoma. The up-regulated expression of CTD-2561J22.5 in a patient with liver cancer is discovered for the first time; and through ROC curve analysis on relevant data, the result that the CTD-2561J22.5 has the higher diagnosis efficiency is discovered. Cell experiments prove that through down-regulating of the expression level of the CTD-2561J22.5, the proliferation activity of liver cancer cells can be reduced; and the result shows that the CTD-2561J22.5 can be used as a molecular target to be applied to the treatment of the hepatocellular carcinoma.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a reagent for detecting and targeting lncRNA biomarkers and its application in hepatocellular carcinoma. Background technique [0002] The incidence of hepatocellular carcinoma (Hepatocellular Carcinoma) is on the rise in the world. As one of the most common malignant cancers in the world, hepatocellular carcinoma has the characteristics of high lethality (Forner A, ReigM, Bruix J[J ]. Hepatocellular carcinoma. Lancet. 2018, 391(10127): 1301-14.). Currently, potentially curative treatments for HCC include liver resection and transplantation, but five-year postoperative survival remains low. The incidence of hepatocellular carcinoma ranks fifth among all malignant cancers. As one of the most common malignant tumors in the world, its mortality rate is extremely high, and the top three are gastric cancer, lung cancer, and liver cancer. In recent years, the incidence of hepatocellular c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11A61K31/7105A61P35/00
CPCA61K31/7105A61P35/00C12Q1/6886C12Q2600/158
Inventor 黄孝伦姚豫桐冯天航赖春友
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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