中文摘要：

目的：探讨RNA干扰对HeLa细胞中血管内皮生长因子（VEGF）表达和细胞增殖的影响,为人宫颈癌的治疗提供理论依据。方法：构建针对VEGF165的RNA干涉质粒,稳定转染HeLa细胞,设HeLa组、对照组和干涉组,用RT-PCR方法检测稳定细胞株中VEGFmRNA的表达,Western blotting和ELISA方法检测VEGF165蛋白的表达,MTT及流式细胞术检测VEGF siRNA作用下的细胞增殖,Hoechst33342染色检测VEGF siRNA作用下细胞凋亡的变化。结果：成功建立了VEGF siRNA的稳定细胞株,与对照组比较,干涉组VEGF的mRNA和蛋白表达水平均明显降低（P〈0.05）。与对照组比较,稳定转染VEGF siRNA的细胞株在24、48和72h的细胞生长抑制率分别为12.8%±5.4%、17.4%±3.7%和27.2%±5.7%,细胞增殖明显受到抑制（P〈0.01）。与对照组比较,Hoechst33342染色显示VEGF siRNA对HeLa细胞的细胞凋亡率没有影响。结论：VEGFsiRNA可以有效抑制HeLa细胞中VEGF mRNA和蛋白的表达,抑制细胞的增殖,提示VEGF在宫颈癌的发生、发展中具有重要作用,抑制VEGF的表达有望成为一种治疗宫颈癌的途径。

英文摘要：

Objective To investigate the effects of small interference RNA （siRNA） targeting vascular endothelial growth factor （VEGF） on VEGF expression and cell proliferation of cervical carcinoma HeLa cells,in order to provide a theoretical basis for the treatment of human cervical carcinoma.Methods siRNAs plasmids targeting VEGF165 were constructed and stably transfected into HeLa cells.HeLa group,Mock group and VEGF siRNA group were set up.The expressions of VEGF mRNA and protein in stable transfectants were confirmed by RT-PCR,Western blotting and ELISA assay,respectively.The cell proliferation was analyzed by MTT and flow cytometry.Apoptosis was measured by Hoechst33342 staining.Results VEGF siRNA stably transfected cell lines were successfully established.The siRNA targeting human VEGF gene effectively decreased the expression of VEGF at RNA and protein levels,and inhibited the proliferation of HeLa cells （P〈0.05）.The inhibitory rates of cell proliferation in HeLa group,Mock group and VEGF siRNA group at 24,48 and 72 h were 12.8%±5.4%,17.4%±3.7%,and 27.2%±5.7%.Compared with Mock group,VEGF siRNA significantly inhibited the cell proliferation in HeLa cells（P〈0.05）.Hoechst33342 staining result showed that VEGF siRNA did not have obvious effect on the apoptotic rate of HeLa cells compared with Mock group.Conclusion The siRNA targeting human VEGF gene could effectively reduce the expression of VEGF both at RNA and protein levels,and inhibit the proliferation of cervical carcinoma cells in vitro.It shows that VEGF plays a role in the development of cervical carcinoma and it may be beneficial in finding new gene therapy for cervical carcinoma.