Effect of shRNA-mediated survivin gene silencing on the biological behavior of human bladder transitional cell carcinoma T24 cells

摘要

To construct the expression vector of the?survivin-specific?short hairpin?RNA (shRNA)?and to investigate?its effect on?the biological behavior of human bladder transitional?cell carcinoma?T24 cells. Survivin-specific?short?oligonucleotides were designed,?synthesized, and?cloned into?the?PTZU6+1?vector, resulting in a?survivin-specific?shRNA?expression construct,?which was then?transfected into?T24?cells.?The mRNA and protein levels of survivin were determined by?semi-quantitative reverse transcription?polymerase?chain reaction?(RT-PCR)?and?Western blot analysis, respectively. Tumor?cell growth,?movement and?neoplasm invasiveness?were detected by a cell?growth curve, a random?motion experiment, and a Matrigel?transmembrane?assay.?Target?DNA?fragments?containingpsurvivin-shRNA vectors was verified by?EcoR I?/?Hind III?double digestion?and sequencing and successfully transfected?into?T24?cells.?Transfection?of?psurvivin-shRNA?cellssignificantly?inhibited?mRNA?levels by 61.73%?and protein?expression?by?73.37%. The ability of cell invasion?and?movement?was?significantly?decreased.?The?inhibition rates of tumor cell growth were?59.13%?on day 3 and?83.86% on day 5.?A survivin-specific?shRNA expression vector was constructed based on the PTZU6 +1?vector and was transfectedinto?T24?cells, which resulted in the inhibition of T24 cell proliferation and movement.