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Use of survivin gene targeted siRNA in preparing medicine capable of regulating and controlling tumour cell cycle

A technology for tumor cells and uses, which is applied in the field of biomedicine and 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

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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 survivin gene targeted siRNA in preparing medicine capable of regulating and controlling tumour cell cycle
  • Use of survivin gene targeted siRNA in preparing medicine capable of regulating and controlling tumour cell cycle
  • Use of survivin gene targeted siRNA in preparing medicine capable of regulating and controlling tumour cell cycle

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Experimental program
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Effect test

Embodiment 1

[0023] siRNA design and synthesis

[0024] Instruments OligoPilot II DNA / RNA Synthesizer (Pharmacia, Sweden), HP-1100 High Performance Liquid Chromatography (Hewlett-Packard, USA), DU2640 UV Spectrophotometer (Backman, USA).

[0025] 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.

[0026] Design of siRNA According to the sequence of the open reading frame (ORF) of the survivin gene, a small molecule interference RNA CPUsiRNA1 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. CPUsiRNA1 sense strand: 5’-ACUGGACAGAGAAAGAGCCdTdT-3’, antisense strand:...

Embodiment 2

[0031] Antitumor Pharmacodynamics Test

[0032] Materials and methods:

[0033] Lung cancer A549, gastric cancer MGC-803, liver cancer Bel7402, breast cancer MCF-7, and leukemia HL60 cell lines were preserved in our laboratory. The transfection reagent Lipofectamin TM 2000 was purchased from Invitrogen, and Taq DNA polymerase, dNTP, DNAMarker and protein marker were 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.

[0034] Cell culture Lung cancer A549, gastric cancer MGC-803, liver cancer Bel7402, breast cancer MCF-7, leukemia HL60 cell lines in RPMI1640 culture medium containing 10% fetal bovine serum, 37 ° C, 5% CO 2 cultured in an incubator.

[0035] 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 44 hours...

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Abstract

The invention relates to the field of biological medicine, in particular to a use of a survivin gene targeted siRNA in preparing medicine capable of regulating and controlling tumour cell division cycle. Experiments indicate that the survivin gene targeted siRNA can change the cell division cycle of lung cancer cell, gastric cancer cell, liver cancer cell, breast cancer cell and leukemia cell, and obviously restrain the cell division of the lung cancer cell, gastric cancer cell, liver cancer cell, breast cancer cell and leukemia cell. The survivin gene targeted siRNA can be used for preparing the medicine capable of restraining the cell division of the tumour 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 division cycle. Background technique [0002] Malignant tumors are currently one of the main killers that threaten human life and health. Although various treatments continue to emerge, no truly effective radical cure has been found so far. Therefore, the treatment of tumors has long been a worldwide problem that has plagued the medical community. Forcing researchers to constantly look for new technologies and methods, trying 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 has two outstanding features: ①It is specifically expressed in almost all kind...

Claims

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Application Information

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IPC IPC(8): C07H21/02A61K48/00A61K31/7088A61P35/00
Inventor 陆云华
Owner YICHUN UNIVERSITY
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