Effect of silencing gene DNA methyltransferase 3b expression by shRNA on proliferation of bladder cancer T24 cells / 中国医师杂志

ABSTRACT

Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.