Application of hsa-novel-circ-0006787 molecule in treatment of hepatocellular carcinoma

A molecular, anti-cancer drug technology, applied in the application field of anti-cancer drug research and development, can solve the problems of accelerated tumor growth, patient injury, etc., and achieve the effect of promoting proliferation and tumorigenicity

Pending Publication Date: 2021-07-16
INNER MONGOLIA MEDICAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for those patients who can be diagnosed and treated after the relevant symptoms appear, the 5-year survival rate is only 14.1%, so it is very important for early screening and prognosis of liver cancer patients
Routine tumor marker screening may need to determine the location of the patient's solid tumor, and perform tissue biopsy through puncture and other methods. It is not suitable for disease screening in healthy people, and it is also more harmful to patients, and adverse stimuli may accelerate tumor growth.

Method used

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  • Application of hsa-novel-circ-0006787 molecule in treatment of hepatocellular carcinoma
  • Application of hsa-novel-circ-0006787 molecule in treatment of hepatocellular carcinoma
  • Application of hsa-novel-circ-0006787 molecule in treatment of hepatocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Real-time fluorescent quantitative PCR (qRT-PCR) detects hsa-novel_circ_0006787 expression

[0032] The biological synthesis and structure diagram of hsa-novel_circ_0006787 is as follows figure 1 As shown, hsa-novel_circ_0006787 is formed by reverse splicing of exons 5, 6, and 7 of RBPMS gene.

[0033] 1. Collection of clinical samples

[0034] Since 2018, our laboratory has collected liver cancer tissues and adjacent normal liver tissues from 31 patients with primary liver cancer from Huaihe Hospital Affiliated to Henan University and Kaifeng Cancer Hospital. Tissue samples were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator. The entire collection and follow-up experiment process conformed to the requirements of medical ethics and strictly followed the principle of confidentiality of case data.

[0035] 2. RNA extraction and real-time fluorescent quantitative PCR (qRT-PCR)

[0036] (1) RNA extraction

[0037] The removed liver t...

Embodiment 2

[0064] Example 2: The lentiviral vector overexpressing hsa-novel_circ_0006787 inhibits the tumorigenicity of liver cancer cells

[0065] 1. Viral Vector Construction

[0066] The cDNA sequence of hsa-novel_circ_0006787 was inserted into the lentiviral expression vector to construct a viral vector overexpressing hsa-novel_circ_0006787. The sequence map of hsa-novel_circ_0006787 was provided by the inventor, and the viral vector was constructed by Shanghai Gemma Technology Co., Ltd.

[0067] 2. Cell lines and cell culture

[0068] Human liver cancer cell line PLC / PRF / 5 cells were taken and cultured in MEM supplemented with 10% fetal bovine serum and 1% double antibody based on 5% CO 2 Incubate at 37°C in an incubator.

[0069] 3. Viral Transfection

[0070] (1) 18-24 hours before lentiviral transfection, the adherent cells were treated with 1×10 5 / well into a 24-well plate. Make the number of cells at the time of lentiviral transfection 2×10 5 / hole left and right.

[...

Embodiment 3

[0097] Example 3: Knocking down hsa-novel_circ_0006787 promotes the tumorigenicity of liver cancer cells to reversely verify that hsa-novel_circ_0006787 can be used as a new target for the treatment of liver cancer

[0098] 1. Small interfering RNA (siRNA) construction

[0099] The specific small interfering RNA (siRNA) targeting the back splicing site of hsa-novel_circ_0006787 and the negative control (si-NC) were synthesized by Gemma Gene (Shanghai), and the sequence map of hsa-novel_circ_0006787 was provided by the inventor. The siRNA target sequence is as follows:

[0100] TGAATACTATGGCATCCGC

[0101] 2. siRNA transfection

[0102] 50 nM siRNA was transfected into liver cancer cells using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). Fluorescent quantitative PCR was used to detect the interference effect.

[0103] image 3 The qRT-PCR results showed that the effect of siRNA knockdown on hsa-novel_circ_0006787 (Panel A) was significant.

[0104] 3. CCK-8 experim...

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Abstract

The invention belongs to the field of biological medicine, and relates to application of an hsa-novel-circ-0006787 molecule in treatment of hepatocellular carcinoma, in particular to application of the hsa-novel-circ-0006787 molecule as a diagnostic marker in treatment of hepatocellular carcinoma or application of the hsa-novel-circ-0006787 molecule as a target in research and development of anti-cancer drugs. The hsa-novel-circ-0006787 molecule disclosed by the invention is a circRNA molecule with low expression in hepatocellular carcinoma tissues, and the nucleotide sequence of the hsa-novel-circ-0006787 molecule is SEQ ID NO. 1. A product for diagnosing the hepatocellular carcinoma comprises a chip, a kit, test paper or a high-throughput sequencing platform and the like. The anti-cancer drug comprises a gene drug and a chemical drug which are used for enhancing the expression of the hsa-novel-circ-0006787.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of hsa-novel_circ_0006787 molecule in the treatment of liver cancer, specifically the application of hsa-novel_circ_0006787 molecule as a diagnostic marker in the treatment of liver cancer or as a target in the research and development of anticancer drugs. Background technique [0002] Primary liver cancer is one of the most common clinical malignant tumors. The annual death toll of liver cancer in my country is 42.2 / 10000, second only to lung cancer and gastric cancer. Commonly used methods for the treatment of liver cancer include surgical resection, liver transplantation, adjuvant therapy, and interventional therapy. Among them, the 5-year survival rate of liver cancer patients diagnosed early and received appropriate treatment or intervention exceeds 50%. However, for those patients who can be diagnosed and treated after the relevant symptoms appear, the 5-year surviv...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113A61K45/00A61K31/7105A61P35/00
CPCC12Q1/6886C12N15/113A61K45/00A61P35/00C12Q2600/158C12Q2600/178C12Q2600/136C12N2310/532C12N2320/32Y02A50/30
Inventor陈卫东叶文凌王凤玲聂小博赵世振侯瑞芳李惟国赵鹏伟
OwnerINNER MONGOLIA MEDICAL UNIV