Pancreatic　cancer　is　a　fatal　human　malignancy　associated　with　an　exceptionally　poor　prognosis.　Novel　therapeutic　strategies　are　urgently　required　to　treat　this　disease.　In　addition　to　immunosuppressive　activity,　triptolide　possesses　strong　antitumor　activity　and　synergistically　enhances　the　antitumor　activities　of　conventional　chemotherapeutic　drugs　in　preclinical　models　of　pancreatic　cancer.　The　present　study　investigated　the　antitumor　effects　of　triptolide　in　pancreatic　cancer　cells,　either　in　combination　with　gemcitabine,　or　alone.　The　pancreatic　cancer　BxPC-3　and　PANC-1　cell　lines　were　treated　with　triptolide,　which　resulted　in　time-and　dose-dependent　growth　arrest.　When　incorporated　into　a　sequential　schedule,　triptolide　synergistically　increased　gemcitabine-induced　cell　growth　inhibition　and　apoptosis,　in　addition　to　the　cooperative　regulation　of　B-cell　lymphoma　2　family　proteins　and　loss　of　mitochondrial　membrane　potential.　Furthermore,　triptolide　enhanced　gemcitabine-induced　S　phase　arrest　and　DNA　double-strand　breaks,　possibly　through　checkpoint　kinase　1　suppression.　The　results　of　the　present　study　suggest　that　triptolide　has　therapeutic　potential　for　the　treatment　of　pancreatic　cancer,　particularly　when　administered　in　combination　with　gemcitabine.