Early biomarkers for pancreatic cancer (PC) detection are required in order to improve patient outcomes. The present study aimed to identify serum biomarkers for PC diagnosis using proteomics, unveil the underlying pathological mechanisms and provide reliable data for the early diagnosis of PC. Isobaric tags for relative and absolute quantification and two-dimensional-liquid chromatography-tandem mass spectometry were used to compare serum samples from patients with PC and healthy individuals. Mascot and Scaffold were used for raw data processing, and Panther for gene ontology (GO) analysis. Igenuity® Pathway Analysis (IPA) was utilized to assess canonical pathways and protein-protein interactions. In total, 76 differentially expressed proteins were identified. The candidate protein DNA repair protein 50 (RAD50) was elevated in patients with PC compared with healthy individuals. In addition, transforming growth factor-β1 (TGF-β1) and apoptotic protease activating factor 1 (APAF-1) were downregulated in PC. GO analysis revealed that the extracellular matrix was increased in PC, as well as receptor function and enzyme regulation; additionally, reduced nucleic acid binding transcription factor activity was observed. IPA analysis demonstrated that the significantly altered canonic pathways were liver X receptor/retinoid X receptor (RXR) activation, the production of nitric oxide and reactive oxygen species in macrophages, the coagulation system, acute phase response signaling and lipopolysaccharide/interleukin-1 mediated inhibition of RXR function. To conclude, RAD50, TGF-β1 and APAF1 are candidate biomarkers for the diagnosis of early PC. The results from the present study could help identify future therapeutic drugs for PC.