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Use of micromolecule RNA-30b-5p as target molecule

A mir-30b-5p, target gene technology, applied in medical preparations containing active ingredients, pharmaceutical formulations, anti-tumor drugs, etc. Growth and Invasion, Reduced Growth and Invasion Ability, and Inhibition of Hepatoma Cell Proliferation

Active Publication Date: 2022-02-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, a large number of studies have confirmed that HBV infection affects the host cell signaling pathway and then affects the normal function of liver cells.

Method used

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  • Use of micromolecule RNA-30b-5p as target molecule
  • Use of micromolecule RNA-30b-5p as target molecule
  • Use of micromolecule RNA-30b-5p as target molecule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Analysis of the difference in transcription level of miR-30b-5p in HBV-related or non-HBV-related hepatocellular carcinoma tissues

[0038] Add Trizol reagent (1 mL per 100 mg of tissue) to the tumor tissues of 7 patients with liver cancer in the experimental group and 7 control groups obtained through surgical resection, grind with an electric homogenizer, and let stand for 5 minutes after completion. Chloroform was added to the tissue homogenate (200 μL per 100 mg of tissue homogenate), shaken vigorously and left to stand for 5 minutes, then centrifuged at 4° C. for 15 minutes (12000 rpm). After centrifugation, transfer the aqueous phase of the layered liquid to another centrifuge tube, add an equal volume of isopropanol and mix well, and let stand at room temperature for 20-30 minutes. Centrifuge again for 10 minutes, discard the supernatant, a white precipitate can be seen at the bottom of the tube, and put the centrifuge tube into an ultra-clean bench to...

Embodiment 2

[0044] Example 2: Inhibiting the level of miR-30b-5p in HBV-related liver cancer cells can inhibit the growth and invasion of tumor cells

[0045] In this example, the inventors first selected a non-HBV-related liver cancer cell line Huh7, and compared the effect of miR-30b-5p on cell growth and invasion ability when infected with or not infected with HBV by scratch test. The miR-30b-5p-specific siRNA was transfected into Huh7 cells or Huh7 cells infected with HBV, and compared with Huh7 cells, respectively. 24 h after transfection, the cell culture dish was scratched with a pipette tip and washed twice with PBS to remove scraped cells. The cells were observed and photographed at 0h, 24h and 48h after the scratch, and the migration area and migration rate of the cells on both sides of the scratch were analyzed and calculated using Image J software. The results are as follows image 3 shown. It can be seen that the distance between cells on both sides of the scratch between t...

Embodiment 3

[0048] Example 3: Inhibition of miR-30b-5p in animal models can inhibit tumor growth

[0049] In this example, miR-30b-5p-specific siRNA was transfected into Hep3B cells, and Hep3B cells were used as a control group. Description: Hep3B is a human liver cancer cell line that has integrated a complete HBV genome, and HBV virus particles can be detected in the supernatant during its growth and proliferation.

[0050] Cells were collected after transfection and resuspended in serum-free DMEM medium to adjust the cell concentration to 2×10 7 cell / mL. The same amount of Hep3B cells and the Hep3B cells transfected with the above siRNA were inoculated subcutaneously into nude mice with a syringe, and three mice were inoculated in each group. After inoculation, the growth of the tumor was observed and the size of the tumor was recorded. The above-mentioned mice were sacrificed 26 weeks after the inoculation, and the size of the tumor was measured after peeling off and photographed fo...

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Abstract

The present invention relates to the use of a micromolecule RNA-30b-5p as a molecular target for resisting hepatitis B virus-related hepatocellular carcinoma. The invention discloses an application of miR-30b-5p as a target molecule in the preparation of anti-HBV-related hepatocellular carcinoma drugs. By inhibiting the transcription of miR-30b-5p or making it lose the function of binding to target genes, the growth and invasion of HBV-related liver cancer cells can be inhibited.

Description

technical field [0001] The invention relates to the use of micromolecule RNA-30b-5p as a molecular target for resisting hepatitis B virus-related hepatocellular carcinoma. Background technique [0002] Primary liver cancer is a disease that seriously threatens human health. It has a high degree of malignancy, is prone to recurrence and metastasis, and has a poor prognosis. It ranks fourth among the causes of cancer death worldwide. Primary liver cancer mainly includes three histopathological types: hepatocellular carcinoma, intrahepatic cholangiocarcinoma, and mixed cell carcinoma, among which hepatocellular carcinoma (HCC) is the most common case type of primary liver cancer one. [0003] Liver cancer can be induced by a variety of reasons, including viral infection, alcohol consumption, obesity, exposure to chemical reagents, and long-term intake of moldy food. In my country, hepatitis, liver cirrhosis and eventually liver cancer caused by chronic infection of hepatitis ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61P35/00A61P1/16A61P31/20
CPCA61K45/00A61P35/00A61P1/16A61P31/20
Inventor 刁宏燕王凯航陈文标姜晶晶章旭君毕珂凡
Owner ZHEJIANG UNIV
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