Objective:To find potential molecular targets for pancreatic cancer diagnosis by comparing and analyzing differential proteins in serum between patients with pancreatic cancer and healthy controls,providing evidence for the early diagnosis of pancreatic cancer. Methods:The protein in serum were separated by two-dimensional gel electrophoresis(2-DE)after the top 14 high abundance proteins were removed from serum with the ProteinExtractTM Removal kit.The significantly differentially expressed proteins were analyzed by matrix-assisted laser desorption-ionization time-of flight mass spectrometry(MALDI-TOF-MS),and some of them were further validated by western blot and immunohistochemistry detection. Results:Protein in serum of each case was successfully separated on 2D gels.Among seventy-eight differently expressed protein,fifty-four proteins were successfully identified by MS with twenty-nine proteins higher expressed in tumors and twenty-six lower expressed in patients. The overexpression of Ras-like without CAAX 1 and Rho-associated protein kinase 2 were further confirmed by western blot and mmunohistochemistry analysis. Conclusion:There were Significant discrepancies in serum i protein expression between patients with pancreatic cancer and healthy controls.And some of the differentially expressed proteins found by the proteomic approach may be potential molecular targets for pancreatic cancer diagnosis.