Objective: our study was to investigate the effect of hepaCAM gene on cell cycle of bladder cancer cell line T24 and BIU-87. Method: Adenovirus vector with hepaCAM was infected into T24 and BIU-87 cells. The expression of hepaCAM mRNA and protein were measured byRT-QPCR and Western-blot. Immunofluorescence detected the cellular localization of hepaCAM. The distribution of cell cycle was analyzed by FCM. The protein expression of cyclinD1 was measured by Westein-blot. Result: The expression of hepaCAM mRNA and protein was increased in two cell lines infected with pAdH5-hepaCAM. The G0/G1 cycle was increased in infected cells, and the protein expression of cyclinD1 was decreased in same cells. Immunofluorescence revealed that hepaCAM protein localized on cytoplasm in well-spread cells, otherwise it localized on the junction of each cell. Conclusion: T24 and BIU-87 cells that infected pAdH5- hepaCAM was blocked in G0/G1 cycle, and it can decreased the protein expression of cyclinD1.