Use of high efficiency siRNA for preparing drug capable of adjusting and controlling tumor cell cycle

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

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 drug capable of adjusting and controlling tumor cell cycle
  • Use of high efficiency siRNA for preparing drug capable of adjusting and controlling tumor cell cycle
  • Use of high efficiency siRNA for preparing drug capable of adjusting and controlling tumor cell cycle

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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), DU 2640 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 According to the sequence of the open reading frame (ORF) of the survivin gene, the small molecule interference RNA CPUsiRNA3 and 1 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 3...

Embodiment 2

[0029] Antitumor Pharmacodynamics Test

[0030] Materials and methods:

[0031] Lung cancer A549, gastric cancer BGC-823, liver cancer Bel7402, breast cancer MDA-MB-435, and cervical cancer Hela cell lines were preserved in our laboratory. The transfection reagent Lipofectamin TM2000 was purchased from Invitrogen, Taq DNA polymerase, dNTP, DNA Marker and protein Marker was 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 BGC-823, liver cancer Bel7402, breast cancer MDA-MB-435, and cervical cancer Hela cell lines were cultured in RPMI1640 medium containing 10% fetal bovine serum in an incubator at 37°C and 5% CO2 .

[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 ...

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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 drug capable of adjusting and controlling the tumor cell cycle. The high efficiency small interfering RNA can silence the expression of survivin, adjust and control the cycle of lung cancer, gastric cancer, liver cancer, breast cancer and cervical carcinoma cells, and obviously inhibit the division of the lung cancer, gastric cancer, liver cancer, breast cancer and cervical carcinoma cells. The small interfering RNA can also prepare the drug for inhibiting the division of the tumor cell.

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] 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 cell mitosis, pa...

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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