Comparative Proteomics Analysis Of Human Cholangiocarcinoma Bile

Background and Objective:At present, cholangiocarcinoma is a common digestive tract tumor in the world and its 5-year survival rate is very disappoing.Because most of the patients are diagnosed at advanced stage,early diagnosis may improve the prognosis of patients with cholangiocarcinoma.In recent years,the fast development of proteomics provides a new technology for detecting tumor markers.This study applies the cross-correlation technique of proteomics to estabilish a 2﹣DE human biliary map for the purpose of comparing differential expressing proteins from cholangiocarcinoma and normal bile,which we can identify biomarkers for early diagnosis of cholangiocarcinoma.Methods:(1)To collect bile samples from cholangiocarcinoma and normal human ,using acetone and TCA/ acetone to extract protein.(2) The proteome in human bile was effectively concentrated and separated by isoelectric focusing on first dimension and SDS-PAGE on second dimension.(3)The 2﹣DE maps were visualized affer siliver staining and analyzed by PDQuest 8.0 2D software to find the differential expression proteins . ( 4 ) Discrepant protein spots were extracted and been enzymolysis and progressing mass spectrogram identification,then according the results of the PMFs to find the discrepant proteins by searching internet using Mascot MS-fit software from SWISS-PROT database.(5)To enquiry the function of Discrepant proteins.Results: (1) Two﹣dimensional protein maps of cholangiocarcinoma and matched normal bile were gained successfully.(2) Acetone- precipitation combined with two-dimensional gel electrophoresis is a suitable approach to study the proteome in human bile.(3) To compare loading different protein quantities,we find 200μg can acquire a 2-DE map better resolving power.(4)5 significant proteins were identified by using mass spectrometry and database,including Angiopoien-related protein 1,Fibroblast growth factor substrate 2 , ?–catenin , Kertain , typeⅠCytoskeletal 16,Ras-related protein 37.Conclusion:(1)Establishment and optimization of the harvest methods for bile samples and 2-DE/MS technique for bile proteomics may be effectively applied in the study of bile proteome and are useful to further study.(2) Proteomic analysis can identify the proteins with variance of cholangiocarcinoma bile versus normal human bile as well as providing probable new biomarkers correlated with biological behavior of cholangiocarcinoma.