Collectively, our results demonstrate that combination of Tanshinone I and TRAIL synergistically induce apoptosis via upregulation of miR135a-3p and DR5 in prostate cancer cells and overexpression of miR135a-3p enhances upregulation of DR5 leading to cell death by combined treatment of TRAIL and Tanshinone I. Thus, our findings suggest that Tanshinone I can be used as a potent sensitizer for TRAIL-based combination therapy in prostate cancer cells. Here, TNFRSF10B is linked to prostate cancer.