Aldo-keto reductases 1C3 (AKR1C3) were highly expressed in docetaxel resistant prostate cancer cell lines after evaluating by gene differential expression microarray. This study aimed to investigate the effects of AKR1C3 silencing on the sensitivity of prostate cancer cells to docetaxel treatment. PC-3 cells were transfected with siRNA against AKR1C3 and treated with docetaxel to study the effect of silencing AKR1C3 on the drug sensitivity of the prostate cancer cells. Expression of AKR1C3 in PC-3 cells of various groups was detected by Western blot. Cell viability was examined in the presence or absence of docetaxel using MTT assay. Results demonstrated that silencing of AKR1C3 significantly promoted docetaxel induced cell growth inhibition in prostate cancer cells. Therefore, AKR1C3 silencing enhances chemotherapy of prostate cancer and could be a potential therapeutic target.