Sirna On Cervical Cancer Siha Cells, Vegf-165 Expression And Cell Proliferation

Objective:To study the effect of siRNA on VEGF-165 expression in vitro on cultured SiHa cells and the exploration of the application of RNA interference on cervical carcinoma gene therapy in the future.Methods:The specific siRNA was transfected into VEGF overexpressed cervical neoplasms cell SiHa lines in vitro by HiPerFect Transfection Reagent.VEGF-mRNA level was detected by real time RT-PCR after transfection with siRNA.The effect of RNA interference was observed at 12,24,48 and 72 hours.The secretion of VEGF protein in the supernatant was evaluated by ELISA.Cell proliferation was tested by MTT assay.Results:Microscopic analysis demonstrated that the optimizing amout of siRNA and HiPerFect Transfection Reagent were 75ng,4.5μl.Sequence specific siRNAs of VEGF-165 effectively downregulated the mRNA levels of VEGF compared with the control(F=99.94,169.27,460.90,115.86,P＜0.05).The protein levels were down regulated obviously post transfection(F=107.06,136.46,147.10,54.25,P＜0.05).The growth of SiHa cells was obviously inhibited(F=70.58,61.17,113.39,100.94, P＜0.05).Whereas the cells in control groups had not obviously changed(P＞0.05).Conelusio:This study demonstrated that specific siRNA can effectively downregulate the expression of VEGF-mRNA and protein,inhibit the cell growth in cervical cancer.The study shows that VEGF play a role in the development of cervical carcinoma and makes the therapeutic approach with gene therapy more possible in cervical carcinoma.