A major obstacle in cancer chemotherapy is the phenomenon of multidrug resistance (MDR), increased P-glycoprotein expression, and abnormal apoptotic processes that may contribute to MDR. Our previous studies demonstrated that JWA is a pro-apoptotic molecule and required for arsenic trioxide and all-trans-retinoic acid-induced cancer cell apoptosis. In this study, the role of JWA in mediating MDR during treatment of choriocarcinoma cells was examined. Data showed that JWA expression was reduced significantly by etoposide (VP16) in JAR MDR cells (JAR/VP16) compared to parent JAR cells. VP16-induced apoptosis in JAR cells was dependent upon the presence of JWA. Knockdown of JWA attenuated VP16-induced apoptosis, and was accompanied by significantly reduced caspase-9 activity and inhibition of JNK phosphorylation. Loss of mitochondrial transmembrane potential induced by VP16 was accompanied by higher JWA expression. JWA was also involved in downregulation of P-glycoprotein through JNK signal pathway. These results suggest that JWA may play an important role in the therapeutic responses to chemotherapeutic agents used to treat choriocarcinoma.