Prostate cancer (PCa) is the most commonly diagnosed male malignancy and the second leading cause of male cancer-related deaths. miR-141 has been demonstrated to be inversely correlated with tumorigenicity. In the present study， we investigated the effect of miR-141 and runt-related transcription factor 1 (RUNX1) on PCa cells. We determined that miR-141 was expressed at a low level and RUNX1 was expressed at a high level in PCa tissues in comparison to that in adjacent normal tissues. Upregulation of miR-141 significantly inhibited cell growth， migration and invasion， and promoted cell apoptosis in PCa cells. Furthermore， miR-141 overexpression suppressed the expression of MMP-2 and MMP-9， and increased the expression of FOXO1 and p21. However， overexpression of RUNX1 could antagonize the effects of miR-141 on PCa cells. Our findings demonstrated that miR-141 could suppress cell growth， migration and invasion and induce cell apoptosis by targeting RUNX1 in PCa cells. Thus， miR-141/RUNX1 play critical roles in the progression of PCa and may be promising targets for the diagnosis and treatment of PCa.