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Use of siRNA in preparing medicament for regulating and controlling tumour cell division cycle

A tumor cell and cycle technology, applied in the field of biomedicine, can solve problems such as loss of resistance

Inactive Publication Date: 2009-06-17
YICHUN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, if the C84A mutation occurs in the only BIR region, survivin will lose its resistance to the apoptosis induced by the chemotherapeutic drug Taxol

Method used

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  • Use of siRNA in preparing medicament for regulating and controlling tumour cell division cycle
  • Use of siRNA in preparing medicament for regulating and controlling tumour cell division cycle
  • Use of siRNA in preparing medicament for regulating and controlling tumour cell division cycle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] siRNA design and synthesis

[0022] Instruments OligoPilot II DNA / RNA Synthesizer (Pharmacia, Sweden), HP-1100 High Performance Liquid Chromatography (Hewlett-Packard, USA), DU2640 UV Spectrophotometer (Backman, USA).

[0023] Materials and Reagents Synthesis column (6.3mL), molecular sieve (3A Sigma), 30HL carrier (lot number 256723), RNA Pac PA100 analysis column (Dionex, USA), 5 kinds of nucleotide monomers (A, U, C, G, dT), acetonitrile, dichloroacetic acid, tetrazole, N2 methylimidazole, except the manufacturers indicated, the rest are products of Pharmacia, Sweden.

[0024]Design of siRNA Based on the sequence of the open reading frame (ORF) of the survivin gene, a small molecule interference RNA CPUsiRNA4 and a pair of negative control siRNA were designed, and their sequences were compared in the human EST database to determine that there were no other gene sequences identical to them. CPUsiRNA4 sense strand 5'-AGCAUUCGUCCGGUUGCGCtt-3', antisense strand 3'-ttUCG...

Embodiment 2

[0029] Antitumor Pharmacodynamics Test

[0030] Materials and methods:

[0031] Lung cancer A549, gastric cancer SGC7901, liver cancer SMMC7721, breast cancer MDA-MB-231, and cervical cancer Hela cell lines were preserved in our laboratory. The transfection reagent Lipofectamin TM 2000 was purchased from Invitrogen, Taq DNA polymerase, dNTP, DNA Marker and protein marker Purchased from Takara Corporation. Rabbit anti-human antibody of Survivin protein, HRP-goat anti-rabbit IgG antibody, and DAB chromogenic kit were purchased from Wuhan Boster Company.

[0032] Cell culture Lung cancer A549, gastric cancer SGC7901, liver cancer SMMC7721, breast cancer MDA-MB-231, cervical cancer Hela cell lines in RPMI1640 medium containing 10% fetal bovine serum, 37°C, 5% CO 2 cultured in an incubator.

[0033] MTT method will 2.5×10 6 / L cells were inoculated in a 96-well plate, 180 μl per well, 3 wells in parallel for each group, and a blank and negative drug control were set up. After ...

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Abstract

The invention relates to application of a micromolecule interference RNA to the preparation of medicines capable of regulating and controlling the tumor cell division cycle. The micromolecule interference RNA can submerge the survivin gene expression, regulate and control the cell cycles of the lung cancer, the gastric cancer, the hepatic cancer, the breast cancer and the cervical carcinoma with high efficiency, and remarkably inhibit the division of the cells of the lung cancer, the gastric cancer, the hepatic cancer, the breast cancer and the cervical carcinoma. The micromolecule interference RNA can be used for preparing the medicines capable of inhibiting tumor cell division.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the use of a small molecular interference RNA (siRNA) targeting the survivin gene in the preparation of drugs for regulating tumor cell cycle. Background technique [0002] RNA interference technology (RNAi) is developed in recent years and is considered to be one of the most powerful tools for studying functional genome. This emerging biotechnology provides new techniques and methods for gene therapy at the molecular level of tumors. RNA interference (RNAi) is a post-transcriptional gene silencing (PTGS) phenomenon mediated by double-stranded RNA (double stranded RNA, dsRNA) dsRNA. By introducing exogenous or endogenous dsRNA into various biological cells, endogenous mRNA can be specifically degraded, thereby causing the repression of corresponding gene transcription. As a simple and effective gene knockout technology, RNAi has been widely used in the research of functional genomics...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/02A61K48/00A61K31/7088A61P35/00
Inventor 陆云华
Owner YICHUN UNIVERSITY
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