Inhibitory effect of small hairpin RNA targeting at VEGF on cervical carcinoma cell line HeLa

OBJECTIVE: To study apoptosis of the tumour cells by RNA interference(RNAi) technology to depress human vascular endothelial growth factor(VEGF) gene expression in HeLa cells,and provide a theoretical basis for the treatment of human cervical carcinoma.METHODS: Short hairpin RNA targeting at VEGF gene was designed and synthesized,which was integrated into plasmid expression vector(PGPU6) with green fluorescent protein(GFP),then transfected into HeLa cells by lipofectamine.The expression of GFP and calculate transfection efficiency was observed by fluorescence microscope.The VEGF mRNA expression in HeLa cells was detected by RT-PCR respectively.Apoptosis was measured by flow cytometry.RESULTS: PGPU6/GFP/Neo-shRNA,a shRNA plasmid expression vector,was successfully transfected into HeLa cells.After 48 hours,The VEGF mRNA expression in HeLa cells transfected with PGPU6-shVEGF1 was decreased to 75.0%.Compared with HeLa group and HeLa-shNC group,the apoptosis of HeLa cells in HeLa-shVEGF1 group was remarkably increased(P0.01).CONCLUSIONS: The PGPU6-shRNA expression vector with GFP constructed can detect the gene transfection more conveniently and do not affect the transcription of U6 promoter.VEGF gene silenced can obviously increase apoptosis of HeLa cells,which may be the new target and efficient way for tumor therapy in the future.