Pancreatic cancer is a highly malignant tumor that is well known for its poor prognosis. It has been reported that aspirin can reduce the risk of various cancers, but the potential role of aspirin and gemcitabine in pancreatic cancer pathogenesis and chemotherapy has not been studied. Therefore, we investigated their synergistic anti-tumor effects and explored the potential molecular mechanisms and biological functions underlying their inhibitory effects on the development of pancreatic cancer in the hope of identifying treatment-related or prognostic biomarkers for the effective treatment of pancreatic cancer.