Use of high efficiency siRNA for preparing tumor migration inhibition drug

A technology of small molecule interference and antisense strands, applied in the field of biomedicine, can solve the problems of loss of resistance and achieve the effect of inhibiting tumor spread and tumor cell migration

Inactive Publication Date: 2009-06-03
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 high efficiency siRNA for preparing tumor migration inhibition drug
  • Use of high efficiency siRNA for preparing tumor migration inhibition drug
  • Use of high efficiency siRNA for preparing tumor migration inhibition drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] siRNA design and synthesis

[0019] Instruments OligoPilot II DNA / RNA Synthesizer (Pharmacia, Sweden), HP-1100 High Performance Liquid Chromatography (Hewlett-Packard, USA), DU 2640 UV Spectrophotometer (Backman, USA).

[0020] 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.

[0021] Design of siRNA According to the sequence of the open reading frame (ORF) of the survivin gene, the small molecule interference RNA CPUsiRNA3 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. CPUsiRNA3 sense strand 5'-AGCAUUCGUCCGGUUGCGCtt-3', antisense strand ...

Embodiment 2

[0026] CPUsiRNA3 changes the adhesion between tumor cells and type IV collagen

[0027] Experimental Materials

[0028] Human gastric cancer cell line MGC-803 Cancer Pharmacology Laboratory, China Pharmaceutical University

[0029] Human breast cancer cell line MDA-MB-231 Cancer Pharmacology Laboratory, China Pharmaceutical University

[0030] Human liver cancer cell line HepG2 Center for New Drug Screening, China Pharmaceutical University

[0031] 24-well culture plate Corning Corporation

[0032] Thincert invasion chamber (pore size 8μm) Greiner

[0033] main reagent

[0034] RPMI1640 medium dry powder Gibco company

[0035] Trypsin Amersco

[0036] MTT Sigma

[0037] main instrument

[0038] Horizontal electrophoresis tank Nanjing University Instrument Factory

[0039] Constant Temperature Oscillator Shanghai Zhicheng Analytical Instrument Manufacturing Co., Ltd.

[0040] Fluorescence Inverted Microscope Leica Corporation

[0041] experimental method

[0042] De...

Embodiment 3

[0053] CPUSIRNA3 inhibits the two-dimensional cell migration ability of tumor cells

[0054] Experimental material and main reagent embodiment 2

[0055] Two-dimensional horizontal cell migration assay

[0056] Referring to the method of Nakayama et al., the scratch damage model was used to analyze the effect of small interfering RNA on the migration ability of tumor cells at the two-dimensional level. The operation steps were as follows:

[0057] (1) digest cells with 0.25% trypsin, press 6×10 4 cells / well were seeded into 24-well culture plates and incubated at 37°C in 5% CO 2 cultivated under conditions;

[0058] (2) The next day, after the cells grow into a uniform monolayer, use a sterile 1000 μL pipette tip to gently draw across the central axis of the well. Wash gently with 37°C preheated PBS for 2 to 3 times to remove floating cell debris, add 37°C preheated medium, and place in the incubator to continue culturing;

[0059] (3) After culturing for 0.5 to 1 hour (u...

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Abstract

The invention belongs to the biological medicine field, in particular relating to the use of a high efficiency small interfering RNA for preparing a tumor migration inhibition drug. The high efficiency small interfering RNA can silence the expression of survivin, change the adhesive force of breast cancer, gastric cancer and liver cancer cells against IV collagen, and inhibit the migrating ability of the breast cancer, gastric cancer and liver cancer cells at two-and three-dimensional levels. The small interfering RNA can also prepare the drug for inhibiting the tumor migration.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the use of a high-efficiency small molecule interfering RNA (siRNA) targeting the survivin gene in the preparation of drugs for inhibiting tumor migration. Background technique [0002] Malignant tumors are currently one of the main killers threatening human life and health, and no truly effective cure has been found so far, which forces researchers to constantly search for new technologies and methods in an attempt to fundamentally overcome cancer. [0003] Survivin is a member of the IAP (inhibitor of apoptosis protein) family first screened and isolated from the human genome library by Aitieric.DC of Yale University in 1997. It is located at 17q25, and the gene is specifically expressed in almost all types of tumor tissues, but almost not expressed in normal tissues; after the gene is highly expressed in tumor cells, it not only promotes tumor cell anti-apoptosis, promotes tumor cel...

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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