Tetramethylpyrazine (TMP) has exhibited various anticancer effects. However, its ability to inhibit proliferation, migration, and invasion of prostate cancer (PCa) PC-3 cells is still unclear. In the present study, different concentrations of TMP were co-incubated with PC-3 cells. The pcDNA-FOXM1 plasmid was transfected into cells before treatment with 500 µg/l TMP. The proliferative, migratory and invasive abilities of PC-3 cells were tested by MTT assay, wound healing assay and colony formation assay. Western blotting was used to investigate the expression of FOXM1. We found that, compared with the control, the proliferative, migratory and invasive abilities of PC-3 cells were decreased after incubation with different concentrations of TMP (P<0.01). The expression of FOXM1 was decreased in TMP-treated PC-3 cells (P<0.01). In addition, overexpression of FOXM1 reversed TMP-mediated inhibition of proliferation, migration and invasion of PC-3 cells. We also found that TMP inhibited PCa growth in vivo in a dose-dependent manner. These results suggest that TMP inhibits PC-3 cell proliferation, migration and invasion by downregulation of FOXM1.