VEGF 2'-O-methylation of siRNA inhibit VEGF expression in bladder cancer

Objectives To investigate the inhibition effect of the chemical modification RNA interfere targeting VEGF on the growth of bladder cancer cell.Methods To construct VEGF siRNA recombinant expression vector:PsilencerTM-VEGF-siRNA.To increase the stability of siRNA,the VEGF siRNA were chemically 2'-O -methyl modificatied.By Lipofectamine 2000,the vector was transfected into bladder cancer cells,By fluorescence microscope,siRNA transfection efficiency,the optimal time point were observed.The level of protein and mRNA of VEGF was detected by Western blot and RT-PCR; Cell proliferation was detected by MTT.Results ①The plasmid expressing siRNA against VEGF was successf? lly constructed.②The best transfection efficiency for the pr-oportion of VEGF siRNA and Lipofectamine 2000 was 2μl ∶ 1 μg,and the cells existed the best in 48 h.③The transfection efiiciency was about (50％ ～60)％ at 48h.The level of protein and mRNA of VEGF were decreased after transfecting PsilencerTM-VEGF-siRNA.④MTT assay on cell proliferation showed:Compared with the control group,the cell proliferation rate of VEGF siRNA and siRNA positive control group was significantly different (P ＜ 0.05).Conclusions Chemical modification of VEGF can effectively inhibit the expression of VEGF in bladder cancer,increase stability,and effectively inhibite the growth of bladder cancer. Key words: Urinary ;  Bladder Neoplsms ;  Vascular Endothelial Growth Factor;  Methylation ;  RNA