关键词: HeLa细胞；血管；内皮生长因子； RNA干涉；细胞增殖

Abstract:

 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.

Key words: HeLa cells;vascular;endothelial growth factor;RNA interference;cell proliferation