Application of RNA and carrier in preparation of product for preventing and/or treating liver cancer

A carrier and product technology, applied in the application field of RNA and carrier in the preparation of prevention and/or treatment of liver cancer products, can solve the problem of no drug treatment, etc., and achieve the effect of low human pathogenicity and wide host range

Active Publication Date: 2013-05-08
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In China, HCC is mostly caused by chronic hepatitis B infection, and

Method used

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  • Application of RNA and carrier in preparation of product for preventing and/or treating liver cancer
  • Application of RNA and carrier in preparation of product for preventing and/or treating liver cancer
  • Application of RNA and carrier in preparation of product for preventing and/or treating liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1, construction of miR-122, recombinant adenovirus vector containing miR-122 encoding gene and adenovirus expressing miR-122

[0057] 1. Acquisition of miR-122

[0058] Synthetic miR-122 was synthesized in Guangzhou Ruibo Biotechnology Co., Ltd.

[0059] The nucleotide sequence of miR-122 is sequence 1 in the sequence listing.

[0060] The negative control RNA of miR-122 was purchased from Guangzhou Ruibo Biotechnology Co., Ltd. (for miRNA mimic Ncontrol#22 product information:

[0061] Basic information: common miRNA mimic Ncontrol#22 for humans, mice, and rats

[0062] Mature miRNA name: cel-miR-239b-5p

[0063] Mature miRNA number: MIMAT0000295

[0064] Mature miRNA sequence: UUUGUACUACACAAAAGUACUG)

[0065] 2. Construction of adenovirus recombinant vector containing miR-122

[0066] The nucleotide sequence of the gene encoding the miR-122 precursor is sequence 3 in the sequence table, the gene encodes the miR-122 precursor, and miR-122 (sequence 1) is ...

Embodiment 2

[0117] Example 2, application of miR-122, recombinant adenovirus vector containing miR-122 encoding gene and adenovirus expressing miR-122

[0118] 1. Application of miR-122 and adenovirus expressing miR-122 in the treatment of HBV transgenic mice

[0119] A. The application of miR-122 in the treatment of HBV transgenic mice

[0120] HBV transgenic BALB / c mice aged 6-8 weeks (purchased from the Liver Disease Center of the 458th Hospital of the Chinese People's Liberation Army); mice with similar body weight, about 16g, were randomly divided into 2 groups, 5 mice in each group .

[0121] miR-122 solution: every 10nM miR-122 was dissolved in 0.1ml of PBS (8g / L NaCl, 0.2g / L KCl, 3.628g / LNa 2 HPO 4 12H 2 0.0.24g / L KH 2 PO 4 and water, pH 7.4), to obtain miR-122 solution;

[0122] Negative control RNA solution of miR-122: every 10nM of miR-122 negative control RNA was dissolved in 0.1ml of PBS (8g / LNaCl, 0.2g / L KCl, 3.628g / L NaCl 2 HPO 4 12H 2 0.0.24g / L KH 2 PO 4 and wa...

Embodiment 3

[0222] Example 3, miR-122, the vector containing the gene encoding miR-122 or the application of the adenoviral vector containing the gene encoding miR-122 in inhibiting HBV replication ability in vitro

[0223] 1. Application of miR-122 in inhibiting HBV replication ability

[0224] HepG2 cells were inoculated into 6-well plates one day before transfection, and the confluence of cells reached about 70% during transfection, and the medium was replaced with opti-MEM (life technologies catalog number: 31985070) one hour in advance.

[0225] miR-122 group: HepG2 cells were co-transfected with miR-122 and plasmid pHBV1.3 using lipo2000 to obtain HepG2 cells / miR-122; the optimal transfection ratio of RNA and pHBV1.3 replicating plasmid was 2.5 in volume and mass ul: 1ug, 4-6 hours the cells were washed three times with PBS, and replaced with 2ml of 10% FBS DMEM full medium.

[0226] Negative control group: the same method was used to co-transfect HepG2 cells with miR-122 negative ...

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Abstract

The invention discloses application of RNA and carrier in preparation of a product for preventing and/or treating liver cancer. The invention provides a recombinant adenovirus carrier which is a recombinant adenovirus carrier formed by inserting at least one encoding gene of miR-122 into a pDC312-cmv; and the miR-122 refers to RNA shown in a sequence 1 in a sequence table or RNA encoded by a sequence 3 in the sequence table. The experiment proves that the miR-122 encoding genes are introduced into the adenovirus carrier, the adenovirus is purified and is used for performing an animal experiment, the miR-122 and the recombinant adenovirus carrier or adenovirus can obviously suppress the liver cancer caused by chronic hepatitis B infection, and a novel liver cancer treatment medicine can be developed.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of an RNA and a carrier in the preparation of a product for preventing and / or treating liver cancer. Background technique [0002] At present, 350 million people in the world are infected with HBV (hepatitis B virus), and about 93 million people in China are chronically infected with HBV. About 15-40% of the infected people may develop life-threatening diseases such as liver cirrhosis and liver failure and hepatocellular carcinoma, chronic HBV infection seriously endangers people's health. Existing treatments for patients with chronic hepatitis B include alpha-interferon (IFN-α) and nucleosides or nucleoside analogs (such as lamivudine, adefovir, and entecavir), interferon-α being the first Obtain clinical drugs for the treatment of hepatitis B virus infection. As an important first-line drug, pegylated interferon-α monotherapy can effectively treat 25-40% of chronic...

Claims

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Application Information

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IPC IPC(8): C12N15/861C12N7/01A61K48/00A61P35/00A61P1/16A61P31/20
Inventor 孟颂东郝军莉李长菲胡俊
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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