1 Bim degradation contributes to cisplatin resistance

Cisplatin is the first l ine chemotherapy for the treatment of several cancers. However, the development of cisplatin resistance represents a major clinical problem and the mechanisms of acquired resistance are not fully understood. Here we show that degradation of the BH3only pro-apoptotic protein Bim plays an important role in cisplatin resistance in ovarian cancer. Specifically, we show that treatment of ovarian cancer cells with cisplatin caused Bim phosphorylation and subsequent degradation and that its degradation is associated with cisplatin resistance. We also show that cisplatin treatment caused the activation of ERK, which correlated with Bim phosphorylation and degradation. By inhibiting ERK phosphorylation with the MEK inhibitor and knocking down ERK expression with siRNA, we show that Bim phosphorylation and degradation were blocked, which suggests that Bim is phosphorylated by ERK and that such phosphorylation is responsible for cisplatin-induced Bim degradation. We show that ERK was activated in cisplatin resistant OV433 cells as compared to their counterpart parental OV433 cells . We also show that Bim was phosphorylated and degraded in cisplatin resistant OV433 cells but not in the parental OV433 cells . Importantly, we show that inhibition of Bim degradation by the proteasome inhibitor MG132 sensitized resistant OV433 cells to cisplatin-induced death. Taken together, our data indicate that degradation of Bim via ERK-mediated phosphorylation can lead to cisplatin resistance. Therefore, these findings suggest that cisplatin resistance can be overcome by the combination of cisplatin and the proteasome inhibitors in ovarian cancer cells .