| Objective:To establish an optimized strategy of two-dimensional gel electrophoresis (2-DE) for proteome analysis of human pancreatic juice (pj) and to analyze the differential expression proteins among pj from patients with pancreatic cancer (PC), chronic pancreatitis (CP) and choledocholithiasis by application of proteomics methods. The study is also for the aim of screening tumor marker candidates and understanding the underlying mechanisms of pc. Methods: Effectiveness of different improved running conditions was compared according to their final results on 2-DE maps. The differential expression proteins related to pc were analyzed by improved 2-DE method and image analysis software and identified by mass spectrometry and database searching. Results:Two-dimensional gel electrophoresis maps with high resolution were obtained and eight differentially expressed proteins were identified. Compared with cp and choledocholithiasis juice, the expression of chain A of a covalent dimer of transthyretin, PRSS 2 preprotein and PLRP 1 in pc juice was found to be up-regulated >2-fold differences(P﹤0.01) while the expressions of chain A of crystal structure of lipid-free human apolipoprotein A-I , chain of human lithostathin, regenerating islet-derived 1 beta precursor, elastase 3B preproprotein and chymotrypsinogen B precursor in pc juice were found to be down-regulated >2-fold differences(P﹤0.01).Conclusions:Two-dimensional gel electrophoresis maps with high resolution and fine reproducibility could be obtained with optimized method. Protein spectra of pj from pc patients, cp patients and choledocholithiasis patients had certain difference and some differential proteins were characterized.
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