Small molecule interfering RNA and its anti-tumor use

A small molecule interference and anti-tumor drug technology, applied in the field of biomedicine, can solve the problems of loss of resistance and achieve the effects of reducing drug resistance, inhibiting tumor angiogenesis, and inhibiting tumor cell division

Inactive Publication Date: 2007-09-19
CHINA PHARM UNIV
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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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  • Small molecule interfering RNA and its anti-tumor use
  • Small molecule interfering RNA and its anti-tumor use
  • Small molecule interfering RNA and its anti-tumor use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] siRNA design and synthesis

[0029] Instruments OligoPilot II DNA / RNA Synthesizer (Pharmacia, Sweden), HP-1100 High Performance Liquid Chromatography (Hewlett-Packard, USA), DU 2640 UV Spectrophotometer (Backman, USA).

[0030] 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.

[0031] 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’-ACUGGACAGAGAAAG AGCCdTdT-3’, antisense stran...

Embodiment 2

[0036] Antitumor Pharmacodynamics Test

[0037] Materials and methods:

[0038] Tumor cell lines A549, HepG2 and HL60 were preserved by our laboratory, transfection reagent Lipofectamin TM 2000 was purchased from Invitrogen Company, Taq DNA polymerase, dNTP, DNA Marker and protein Marker were purchased from Takara Company. Rabbit anti-human antibody of Survivin protein, HRP-goat anti-rabbit IgG antibody, and DAB chromogenic kit were purchased from Wuhan Boster Company.

[0039] Cell culture tumor cell lines A549, HepG2 and HL60 in RPMI1640 medium containing 10% fetal bovine serum, 37°C, 5% CO 2 cultured in an incubator.

[0040] 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, discard the supernatant, add 20 μL of freshly prepared 5g / L MTT to each well, incubate for 4 hours, discard the supernatant, add 150 μL DMSO to dissolve, mix we...

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Abstract

The invention relates to biological medicine field, concrete relates to a broad-spectrum small interfering RNA for survivin gene. The medicine designing double chain siRNA for function conservation of survivin gene open reading frame (ORF) has function of broad-spectrum antitumor and inhibition cariers of lung cancer, liver cancer, leukaemia, gastric cancer, cervical cancer, KB and so on, sense strand of the sequence combines with mRNA of survivin gene to disturb interprete thereof to induce apoptosis of tumour cells, to repress abruption thereof, to reduce drug fastness thereof, to repress generation of tumour vein to treatment tumour.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a small molecular interference RNA (siRNA) targeting the apoptosis inhibitor survivin gene, and its anti-tumor application. 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] The apoptosis suppressor gene survivin is an important tumor-related gene discovered in 1997, which was screened and cloned in the human genome library by Ambrosini et al. (.Nature Med, 1997, 3(8): 917-921). Located at 17q25, this gene has two outstanding features: ①It is specifically expressed in almost all kinds ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11A61K31/7088A61K48/00A61P35/00C07H21/02C12N15/113
Inventor 奚涛陆云华邢滢滢丁黎
Owner CHINA PHARM UNIV
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