In order to identify the antitumor effect of TAT-Apoptin on the human bladder cancer EJ cell line and study its impact on the expression of the apoptosis-related genes bax, bcl-2, caspase-3 and survivin, the MTT assay, real-time quantitative PCR and western blot analysis were used in this study. The results of the MTT assay indicated that TAT-Apoptin was able to inhibit the proliferation of EJ cells in a dose-dependent manner. The expression of Bax, Bcl-2, Caspase-3 and Survivin mRNA and protein following the treatment of the EJ cells with TAT-Apoptin (0.1, 0.5, 1, 10, 50 and 100 μg/ml) for 24, 48 and 72 h was analyzed. Cell proliferation was significantly different after treatment with the various concentrations of TAT-Apoptin and the durations of treatment. The level of expression of Bcl-2 and Survivin in EJ cells decreased significantly, while that of Bax and Caspase-3 increased significantly at the mRNA and protein levels.