Prostate cancer (PC) is one of the most common cancers in males. MicroRNAs (miRNAs) are demonstrated to be involved in prostate cancer development and progression. Recently, miR-96 was identified to play a tumor promoting role in several tumors including PC, however, the underlying function of miR-96 in PC still need to be known. In the study, our results demonstrated that miR-96 was higher in prostate cancer tissues compared with adjacent normal tissues. Higher miR-96 was association with higher PSA level, lymph node metastasis, pathologic stage and distant metastasis in prostate cancer patients. Lose-of-function studies showed that down-regulated expression of miR-96 inhibited cell proliferation and cell cycle by regulating down-regulating CyclinA1, CDK2 and CDK4 expression in PC cells. Furthermore, we found that FOXF2 was a target of miR-96 in PC cells and miR-96 promoted cell proliferation by suppressing FOXF2 expression. Thus, these results showed that inhibition of miR-96 may be a target for prostate cancer treatment.