Objective    To construct the plasmid containing short hairpin RNA (shRNA) of survivin to suppress the expression of endogenous survivin gene in T24 cells.     Methods    The recombinant plasmid  pshRNA-survivin2 expression construct was confirmed by EcoR Ⅰ and HindⅢ double digestion and by sequencing. The plasmid pshRNA-survivin2 was stably transfected into transitional cell carcinoma of the bladder (TCCB) cells T24. Then survivin mRNA and protein expressions in the transfected T24 cells were detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. The biological effects of the plasmid pshRNA-survivin2 on T24 cells were evaluated through the detection of their anchorage-independent growth and in vitro invasion by cell migration assay and tranwell chamber assay.     Results    The expression of survivin in transfected T24 cells was markedly depressed at both mRNA and protein levels (61.73% and 73.37%

respectively) as compared with control. Anchorage-independent growth assay showed the growth of tumor cells was retarded. The inhibitory percentage of transfected T24 cells was respectively 59.13% and 83.86% on the third day and the fifth day. Transwell chamber assay showed the invasion ability of T24 cells was inhibited significantly.     Conclusion    The pshRNA-survivin2 expression plasmid

constructed from plasmid pTZU6+1

can be successfully transfected into TCCB cell line T24 and can effectively inhibit the expression of survivin mRNA and protein. Therefore

the proliferation

migration and invasion of T24 cells transfected with pshRNA-survivin2 expression plasmid are inhibited. RNAi targeting survivin has a potential value in gene therapy of bladder cancer.