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Effect Of Small Interference RNA On E6, E7 Gene Of Human Papillomavirus Type-18 In Cervical Cancer Cells

Posted on:2007-03-27Degree:MasterType:Thesis
Country:ChinaCandidate:P WangFull Text:PDF
GTID:2144360182493546Subject:Obstetrics and gynecology
Abstract/Summary:PDF Full Text Request
Objective To study the effect of small interference RNA on E6, E7mRNA of human papillomavirus type-18(HPV18) in cervical cancer cells and its therapy action to cervical cancer.Methods The specific HPV18E6 and E7 siRNA synthesized by in virtro transcription. The cell activity was detected by methyl thiazolyl tetrazolium(MTT) assay to determine the most suitable concentration, then cells were transfected into HPV18 cervical cancer cells by oligofectamine. HPV18E6, E7 mRNA level was examined by semi-quantitative RT-PCR, and the cell cycles were analyzed by flow cytometry.Results The activity of the HeLa cells had an obvious suppression after transfection to 0.57±0.05 and 0.62±0.04 compared with the contrast of 0.87±0.05. The expression of E6 mRNA level was 0.63±0.04 before transfection, decreased to 0.53±0.04, 0.46±0.02, 0.56±0.03 evidently after transfection at 24, 48, 72 hours;The expression of HPV18E7 mRNA level was 0.66±0.03 before transfection and decreased to 0.60±0.05, 0.52±0.04, 0.59±0.02 after transfection at 24, 48, 72 hours;The number of cells in G2 phase was increased after HPV18E6siRNA transfection from (1.4±1.2) % to (66.9±3.5) % , S phase was declined from (39.4±0.4) % to (0±5.5) %, and they were (47.2±0.5 ) % and (5.6±4.2) % in E7 group.Conclusions RNAi exits in Hela cells, and it's effect has specificity.
Keywords/Search Tags:small interference RNA, HPV18, cervical cancer, Hela cells
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