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The prognostic role of  CD 44v9, a variant isoform of  CD 44 and a new cell surface marker of cancer stem cells, remains unclear in bladder cancer ( BC ) patients. Furthermore, limited information is available on the functional role of sulfasalazine ( SSZ ), which could modulate the  CD 44v9‐ xCT  system in order to enhance cisplatin ( CDDP )‐induced cytotoxicity and inhibit the metastatic potential of  BC .  CD 44v9 protein expression was examined immunohistochemically in 63 muscle invasive  BC  ( MIBC ) patients who underwent radical cystectomy.  CD 44v9 expression was independently associated with disease recurrence and cancer‐specific death in  MIBC . Cytotoxic effects, glutathione levels, and reactive oxygen species production by  SSZ  and  CD 44v9 and phospho‐p38  MAPK   protein expression by  SSZ  with or without  CDDP  were assessed in  MBT ‐2V cells with highly metastatic potential. Sulfasalazine exerted cytotoxic effects against  MBT ‐2V cells by inhibiting glutathione levels and inducing the production of reactive oxygen species. Sulfasalazine in combination with  CDDP  appeared to exert strong cytotoxic effects against  MBT ‐2V cells by inhibiting  CD 44v9 expression and upregulating phospho‐p38  MAPK   expression. The inhibitory effects of  SSZ  with or without  CDDP  were also investigated using an  MBT ‐2V lung metastatic model. In the murine lung metastatic  BC  model,  SSZ  significantly prolonged animal survival. Furthermore, the combination of  SSZ  with  CDDP  exerted stronger inhibitory effects on the establishment of lung tumor nodules than  SSZ  or  CDDP  alone.  CD 44v9 expression could be a clinical biomarker for predicting poor outcomes in  MIBC  patients. Sulfasalazine in combination with  CDDP  has potential as a novel therapy against metastatic  BC .

Sulfasalazine could modulate the CD 44v9‐ xCT system and enhance cisplatin‐induced cytotoxic effects in metastatic bladder cancer