MicroRNAs have been reported to be associated with chemosensitivity of several types of cancers. However, the underlying molecular mechanisms are poorly understood. In this study, weexplored miR-218 increased the chemosensitivity to cis-diaminedichloroplatinum treatment of prostate cancer. We found that the expression level of miR-218 was down-regulated in the human prostate cancer specimens. Moreover, overexpression of miR-218 inhibited cell viability, migration, and invasion in PC3 and DU145 cells. Furthermore, we demonstrated that the tumor suppressive role of miR-218 was mediated by negatively regulating branched-chain amino acid transaminase 1 (BCAT1) protein expression. Importantly, overexpression of BCAT1 decreased the chemosensitivity to CDDP treatment of PC3 and DU145 cells. Our study is the first to identify the positive role of miR-218 in chemosensitivity, which will facilitate the development of novel therapeutic strategies for prostate cancer in the future.