Inhibition Of Expression Of VEGF In SGC-7901 Cells By SiRNAs

ObjectiveThis study was to evaluate the effect of small interference RNA (siRNA) on VEGFgene expression. and effect of RNAi-mediated VEGF gene silencing on proliferation andapoptosis of human cancer cell SGC-7901.MethodsDesigned and synthesized small interfering RNA (siRNA) targeted VEGF, which weretransfected into human gastric cancer cell SGC-7901 with lipofectamine. Two siRNAs andone scrambled siRNA were designed on line and obtained by in vitro transcription.the non-transfected cells, Cells treated with lipofectimine and cells treated with SCR were taken ascontrols.MTT was used to detect the inhibitive rate of cells growth,and Hoechst33258 stainingwas used to observe the apoptositic bodies. Flowcytometry method was used to detect thechange of cycling of cell, The inhibitive effect of siRNA on VEGFmRNA level weredetected by RT-PCR, VEGF protein in the supernatant was detected by ELISA.ResultsThe small interfering RNA (siRNA) targeting human VEGF effectively inhibited thesecretion of VEGF in SGC-7901, whereas the control scramble siRNA showed no effects.MTT assay showed that siRNA₁ and siRNA₂ markedly inhibited the proliferation ofgastric Cancer cell SGC-7901, while the control scramble siRNA showed no effects.Hoechst33258 staining showed siRNA induced cell apoptosis and the apoptositic bodiesalso appeared. It affected the distribution of cell cycle, increased cell population in G₀/G₁phase and decreased it in S phase,the expression of VEGF mRNA was significantlyinhibited in SGC-7901. the secretion of VEGF protein in the supernatant of the SGC-7901cells which were examined with ELISA and transfected with siRNA₁ and siRNA₂ for 24hours were significantly lower than the untransfected cells, while the control scramblesiRNA showed no effects.ConclusionssiRNAs targeted to VEGF mRNA in vitro transcripted can effectively inhibit the expressionof VEGF, inhibit proliferation and induce apoptosis of SGC-7901 cells, siRNA moleculemay create a more promising and effectious siRNA-based gene drugs against tumors.