Objective To establish a multidrug resistance cell line of human bladder tumor and study its characteristics and mechanism of muhidrug resistance. Methods Human bladder tumor cell line T24 was induced to muhidrug resistance cell line T24/ADM by intermittent administration of high dose ADM. The multidrug resistance to multianticancer agents of the ceils was evaluated by MTT assay; the expression of MDR gene was assessed by RT-PCR and P-gp expression was determined by flow cytometry. Results T24/ ADM resisted to many anti-tumor agents, and its IC50 of ADM was 16.3 times higher than that of T24. Significant overexpression of MDR gene and p-gp of the multidrug resistant cells were detected. Conclusion T24/ADM is a human muhidrug-resistant cell line, and it's drug resistance relates to the overexpression of MDR gene and p-gp.