Abstract: Objective To establish a cisplatin resistant bladder cancer cell line and to analysis its biological characteristics．Method Drug concentration gradient increasing method was adopted to induce drug resistance. T24 and T24/DDP were observed under phase contrast microscope. Multidrug resistance of the two cell lines was determined by MTT. Cell proliferation was evaluated according to the growth curve. Flow cytometry was performed to determine cycles of the two cell lines. The expression of multidrug resistance related gene multidrug resistance-associated protein (MRP) was detected by qRT-PCR. Results After 14 months, cisplatin-resistant human bladder cancer cell line T24/DDP was successfully established from its parental cell line T24. Two kinds of cells were significantly different from each other by a microscope. T24/DDP cell was 9.51 times more resistant to DDP than T24. T24/DDP cell line exhibited the characteristics of cross-resistance to cisplatin, adriamycin, methotrexate, vincristine. The cell doubling time of T24/DDP was longer than parental cell line T24. Compared with T24， the proportion of G2 and S phase cells in T24/DDP decreased significantly，while that of G1 phase cells increased significantly．The expression of drug-resistance related gene MRP was significantly up-regulated in T24/DDP（P＜0．05）．Conclusion With the characteristics of multidrug resistance, cisplatin-resistant human bladder cancer cell line T24/DDP could be used as a good experimental model for further study on bladder cancer multidrug resistance.