Objective Gemcitabine, the only approved drug for the treatment of pancreatic cancer, is not very effective. Novel and effective cancer chemopreventive agents are urgently needed. Recently, emerging studies determined resveratrol possessed anticancer effects on various cancer cells. We explored the anticancer effect of resveratrol in pancreatic cancer cells and investigated the involved moleculars of action. We also examined whether resveratrol enhanced antitumor activity of gemcitabine in vitro.Methods Proliferation inhibition was assessed by cell count kit-8 assay. Cell cycle phase distribution and apoptotic cells were measured by flow cytometric analysis. We determined the expression of bcl-2, cyclinD1, and activation of caspases-3 and poly(ADP-ribose) polymerase1 proteins used Western blot analysis.Results Resveratrol inhibited the proliferation of three pancreatic cancer cell lines in a dose dependent fashion, and induced accumulation of cells at the G1 phase as well as apoptosis. Our data also demonstrated that resveratrol enhanced gemcitabine-induced apoptosis in pancreatic cancer cells. In addition, resveratrol inhibited the expression of cyclinD1, bcl-2, and induced activation of caspase-3 and poly(ADPribose) polymerase1. Conclusion Our results suggested that resveratrol might be not only a potential regimen, but also an effective chemosensitizer for the chemotherapy of pancreatic cancer.