Investigation Of Proliferation Suppressing And Apoptosis Inducing Effects Of RNAi Targeted To Survivin On TCCB

Objective To investigate the proliferation supperring and apoptosis inducing effects of RNAi targeted to Survivin on T-24 cells, a bladder cancer cell line.Methods An siRNA eukaryotic expression vector targeted to Survivin, pTZU6+1-shRNA-survivin, was emplified in system and identitified by sequencing method. It was transfected into T24 cells following lipofectamineTM2000 protocols. The changes of transcriptional level of survivin gene were detected by semi-quantitive RT-PCR.The effects of proliferation suppressing on T24 cell were detected by MTT assay. The effects of apoptosis inducing on T-24 cells were detected by flow cytometry assay.Rusult The recombined vector was successfully emplified, and was proved to have no mutation sites by sequencing. 24 hours after trancfection the transcriptional level of Survivin gene in T24 cells was significently suppressed at the rate of 41.77%, detected by semi-quantative RT-PCR. The proliferation suppressing effects of RNAi targeted to survivin were observed by MTT assy, 48 hours after transfecion. The proliferation inhibitory rate reached to 52.53%. Meanwhile, 24h after transfection, it was observed that silence survivin by RNAi could significently induce the spontaneous apoptosis of T-24 cells at the rate 16.76%±1.31 , detected by flow cytometry assay and comfirmed by electron microscope.Conclusion Silencing exogenous Suvivin by RNAi can significantly suppress the proliferation of T-24 cells and induce spontaneous apoptosis. RNAi targeted to survivin has a potential role in gene therapy of bladder cancer.