Effects of siRNA on VEGF expression and cell proliferation of HeLa cells
Effects of siRNA on VEGF expression and cell proliferation of HeLa cells
复制标题
siRNA对HeLa细胞VEGF表达及细胞增殖的影响
DOI：
--
复制
发表时间：
2009
影响因子：
--
通讯作者：
Wang J.
中科院分区：
文献类型：
--
作者：
Zuo D.;Li X.-R.;Wang L.;Wang J.
目的探讨针对血管内皮生长因子(VEGF)的小干扰RNA(SiRNA)对宫颈癌HeLa细胞中血管内皮生长因子(VEGF165)表达和细胞增殖的影响，为宫颈癌的治疗提供理论依据。方法构建针对VEGF165的siRNA载体，并稳定地将其导入HeLa细胞，分别设HeLa组、Mock组和VEGFsiRNA组。RT-PCR、Western blotting和ELISA法检测稳定转染组中VEGFmRNA和蛋白的表达。结果成功建立了稳定表达人血管内皮生长因子siRNA的细胞株。针对人血管内皮细胞生长因子基因的siRNA在RNA和蛋白水平上有效地降低了血管内皮生长因子的表达，抑制了HeLa细胞的增殖(P<0.05)。HeLa组、Mock组和VEGFsiRNA组在24、48和72h的细胞增殖抑制率分别为12.8%±5.4%、17.4%±3.7%和27.2%±5.7%。与Mock组相比，HeLa组、Mock组和VEGFsiRNA组的细胞增殖抑制率分别为12.8%±5.4%、17.4%±3.7%和27.2%±5.7%。Hoechst33342染色结果显示，与模型组相比，血管内皮生长因子siRNA对HeLa细胞的凋亡率无明显影响。结论靶向人血管内皮生长因子基因的siRNA能从RNA和蛋白水平有效地抑制宫颈癌细胞的增殖，提示血管内皮生长因子在宫颈癌的发生发展中起一定作用，有望为宫颈癌的基因治疗提供新的思路。
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 (P0.05).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(P0.05).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.