An integrated approach for comparative proteomic analysis of human bile reveals overexpressed cancer-associated proteins in malignant biliary stenosis

Article Article scientifique Article An integrated approach for comparative proteomic analysis of human bile reveals overexpressed cancer-associated proteins in malignant biliary stenosis eng 2014 author Lukic Natalija 702034 author Visentin Rémy 791702 author Delhaye Myriam author Frossard Jean-Louis 35038 author Lescuyer Pierre 0000-0001-6986-8738 562350 author Dumonceau Jean-Marc 565160 author Farina Annarita 649783 1026-1033 10.1016/j.bbapap.2013.06.023 23872482 0006-3002 Proteomics is a key tool in the identification of new bile biomarkers for differentiating malignant and nonmalignant biliary stenoses. Unfortunately, the complexity of bile and the presence of molecules interfering with protein analysis represent an obstacle for quantitative proteomic studies in bile samples. The simultaneous need to introduce purification steps and minimize the use of pre-fractionation methods inevitably leads to protein loss and limited quantifications. This dramatically reduces the chance of identifying new potential biomarkers. In the present study, we included differential centrifugation as a preliminary step in a quantitative proteomic workflow involving iTRAQ labeling, peptide fractionation by OFFGEL electrophoresis and LC-MS/MS, to compare protein expression in bile samples collected from patients with malignant or nonmalignant biliary stenoses. A total of 1267 proteins were identified, including a set of 322 newly described bile proteins, mainly belonging to high-density cellular fractions. The subsequent comparative analysis led to a 5-fold increase in the number of quantified proteins over previously published studies and highlighted 104 proteins overexpressed in malignant samples. Finally, immunoblot verifications performed on a cohort of 8 malignant (pancreatic adenocarcinoma, n=4; cholangiocarcinoma, n=4) and 5 nonmalignant samples (chronic pancreatitis, n=3; biliary stones, n=2) confirmed the results of proteomic analysis for three proteins: olfactomedin-4, syntenin-2 and Ras-related C3 botulinum toxin substrate 1. This article is part of a Special Issue entitled: Biomarkers: A Proteomic Challenge. Adenocarcinoma/complications/metabolism Aged Aged, 80 and over Bile Duct Neoplasms/complications/metabolism Bile Ducts, Intrahepatic/metabolism/pathology Cholangiocarcinoma/complications/metabolism Cholestasis/diagnosis/etiology/metabolism Chromatography, Liquid Cohort Studies Female Humans Immunoblotting Male Middle Aged Pancreatic Neoplasms/complications/metabolism Pancreatitis, Chronic/complications/metabolism Proteomics/methods Tandem Mass Spectrometry Tumor Markers, Biological/metabolism Dewey 576 Dewey 616 136 Département de biologie structurale et bioinformatique 21E 56 Département de médecine 221 false 211 Auto-immunité, inflammation et athérosclérose 270 Biochimica et biophysica acta 1844 5 34a39b1c-2d58-4e15-b5ab-2d17c80378b3 Article (Published version) Restricted 1-s2.0-S1570963913002616-main.pdf 788566 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de biologie structurale et bioinformatique Faculté de médecine / Section de médecine clinique / Département de médecine

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An integrated approach for comparative proteomic analysis of human bile reveals overexpressed cancer-associated proteins in malignant biliary stenosis	Biochimica et biophysica acta			2014	749	6

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Lukic, Natalija