We previously demonstrated that resveratrol (Res) regulates the expression of PKC α and δ, to eventually inhibit growth and induce apoptosis in human gastric cancer cells. In the present study, the effect of Res on the growth of human pancreatic cancer cells was investigated, to further unveil the underlying mechanism. The human pancreatic cancer cell line MIA PaCa-2 was treated with three different concentrations of Res. Cell proliferation was assessed by the MTT assay, and apoptosis was detected by flow cytometry. Reverse transcription-quantitative PCR (RT-qPCR) and western blot analysis were used to measure the mRNA and protein levels of the Hedgehog (Hh) signaling proteins Ihh, Ptch and Smo. Our results revealed that Res can inhibit the cell proliferative ability in a time- and dose-dependent manner. The number of formed colonies in the Res- and 5-Fu (positive control)-treated groups was decreased as compared to the negative control group. Res further induced apoptosis of MIA PaCa-2 cells in a dose-dependent manner. In addition, the levels of Ihh, Ptch and Smo were decreased by Res treatment. Our findings suggest that Res inhibits proliferation and induces apoptosis of MIA PaCa-2 pancreatic cancer cells in vitro, which may be related to its inhibitory effect on the Hh signaling pathway.