Objective: miRNAs, altered expression manner in prostate cancer, have involved in cancer development by regulating proliferation, differentiation, invasion, metabolism and apoptosis. Recently, it has been found that miRNA-627 might mediate colorectal cancer cells as a tumor-suppressor. However, its role on prostate cancer cells remains unknown. Methods: Following transfection with miRNA-627, miRNA-627 expression, cell proliferation, cell cycle, cell apoptosis, migration assay, and real-time PCR analysis were performed in prostate cancer cells. Results: It was found that miRNA-627 inhibited cell proliferation, retarded cell cycle and cell migration in prostate cancer cells. Additionally, miRNA-627 promoted cell apoptosis and upregulated the mRNA levels of MAP3K1, PTPRK and SRA1. Conclusion: Collectively, miRNA-627 interrupts prostate cancer development as suppressing cell proliferation, migration and promoting cell apoptosis and tumor suppressor genes expression, which might provide promising therapeutic effect in the treatment of prostate cancer.