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A kind of long chain non-coding rna Lnc-EPC1-4 and its application

A long-chain non-coding and tumor technology, applied in the field of long-chain non-coding RNA Lnc-EPC1-4, can solve the problems of unclear lncRNA expression and achieve the effect of treatment and prevention

Active Publication Date: 2022-05-03
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the expression of lncRNAs in the serum of patients with early-stage HCC remains unclear

Method used

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  • A kind of long chain non-coding rna Lnc-EPC1-4 and its application
  • A kind of long chain non-coding rna Lnc-EPC1-4 and its application
  • A kind of long chain non-coding rna Lnc-EPC1-4 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Purification of exosomes from serum for quantitative analysis of Lnc-EPC1-4 RT-PCR

[0035] (1) Whole blood was collected from 45 healthy individuals (normal) and patients with hepatitis (hepatitis), cirrhosis (cirrhosis) and HCC (liver cancer), and serum was separated from the whole blood;

[0036] (2) Serum was ultracentrifuged (Beckman L8 ultracentrifuge) at 100,000 g for 20 hours at 10°C. Discard the supernatant and resuspend the exosome-containing pellet in PBS.

[0037] (3) Determine the phenotype of exosomes. The size of the collected exosomes was verified by nanoparticle tracking analysis (NTA) (Particle Metrix, Germany), and the membrane protein content (including CD63 and TSG101) was verified by Western blotting.

[0038] (4) Use a commercial RNA extraction kit to extract the RNA in exosomes, then use a commercial RT-PCR kit to quantitatively analyze the RNA of various tissues, and the PCR primers used to detect the expression content of Lnc-EPC1-4...

Embodiment 2l

[0046] Example 2 Suppression of lnc-EPC1-4 and Construction of Overexpression Cell Lines

[0047] 1. Construction of lnc-EPC1-4 suppression and overexpression system

[0048] (1) Design and synthesize lnc-EPC1-4 siRNA

[0049] siRNA-1: 5'-GCACTAATGGGAGCTCTGT-3';

[0050] siRNA-2: 5'-GAAGCAACATCAAGATGAA-3';

[0051] siRNA-3: 5'-GAGAAAGACAGTAGGTCAA-3';

[0052] Synthesized by Ruibo Biotechnology Co., Ltd. (Guangzhou, China), according to the manufacturer's instructions, the vector and siRNA were transfected into cells using Lipofectamine 2000 (Thermo Fisher, USA) to obtain lnc-EPC1-4 inhibited cell line.

[0053] (2) Amplify lnc-EPC1-4 primer:

[0054] F: 5'-AAGTGTGGCCAGAGAAGAAGAC-3';

[0055] R: 5'-TCAGTGGCAGTTGTTTCAGGTT-3';

[0056] Full-length lncRNA was amplified by PCR using Thermo Scientific Phusion Flash High-Fidelity PCR Master Mix (Thermo), and then the corresponding cDNA was subcloned into pcDNA3.1(+) (Genepharma, China) according to the manufacturer's instruction...

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Abstract

The invention discloses a long-chain non-coding RNA Lnc‑EPC1‑4 and an application thereof. The nucleotide sequence of the Lnc-EPC1-4 is shown in SEQ ID NO: 1; the present invention finds that the lnc-RNA is all lowly expressed in the serum of liver cirrhosis or liver cancer patients, and can be detected by detecting the lnc-RNA in the patient's serum The content of EPC1‑4 is used for screening and detection of liver cancer, especially in the early stage (such as liver cirrhosis stage); at the same time, it was found that lnc‑EPC1‑4 is a tumor suppressor factor, and overexpression of lnc‑EPC1‑4 can promote the apoptosis of liver cancer cells And inhibit its proliferation, so as to realize the treatment and prevention of liver cancer, and provide new ideas and strategies for the prevention and treatment of tumors.

Description

technical field [0001] The present invention relates to the technical field of tumor molecular biology, and more specifically relates to a long-chain non-coding RNA Lnc-EPC1-4 and its application. Background technique [0002] Primary liver cancer (HCC) is a common malignant tumor, its incidence rate and mortality rate account for the 5th and 3rd among malignant tumors in the world, and the 3rd and 2nd among malignant tumors in my country. The death rate in rural areas has risen to the first place. Generally speaking, hepatitis is the early stage of liver cancer, and gradually develops into liver cirrhosis, and finally develops into liver cancer. [0003] In the early stage of HCC, surgery is an effective treatment; however, with the current detection level, most patients develop intrahepatic and extrahepatic metastases shortly after diagnosis, which limits the treatment of radiotherapy and chemotherapy. Therefore, early diagnosis and intervention treatment of HCC are very ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12Q1/6886C12N15/11
CPCC12N15/1135C12Q1/6886
Inventor 姚志成贾昌昌张琪
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV