title: Proteomic analysis of a rat pancreatic stellate cell line using liquid chromatography tandem mass spectrometry (LC-MS/MS).
abstract: Pancreatic stellate cells (PaSC) are emerging as key mediators in chronic pancreatitis and pancreatic cancer pathogenesis. Proteins regulating the biomolecular pathways involved in the conversion of quiescent to activated PaSC may have a significant influence on the development of chronic pancreatitis. We aim to compare differentially expressed proteins in activated and serum-starved non-proliferating PaSC using a mass spectrometry-based proteomics strategy. We cultured an immortalized rat PaSC cell line in media supplemented with 10% fetal bovine serum and in serum-free media. Using gel-based mass spectrometry (GeLC-MS/MS), we identified nearly 1500 proteins. Qualitative and quantitative proteomic analysis revealed several hundred proteins as differentially abundant between the two cell states. Proteins of greater abundance in activated PaSC included isoforms of actin (e.g., smooth muscle actin) and ribosomal proteins. Conversely, proteins more abundant in non-proliferating PaSC than in activated PaSC included signaling proteins MAP kinase 3 and Ras-related proteins. In addition, we have determined the molecular functions and biological pathways for these proteins. We are confident that the application of mass spectrometry-based strategies, such as that described herein, to investigate specific proteins in PaSC may lead to a better understanding of the molecular mechanisms involved in pancreatic diseases, such as chronic pancreatitis.
authors.affiliation: Department of Pathology, Children's Hospital Boston, Boston, MA 02115, USA.
authors.forename: Joao A
authors.initials: JA
authors.lastname: Paulo
authors.forename: Raul
authors.initials: R
authors.lastname: Urrutia
authors.forename: Peter A
authors.initials: PA
authors.lastname: Banks
authors.forename: Darwin L
authors.initials: DL
authors.lastname: Conwell
authors.forename: Hanno
authors.initials: H
authors.lastname: Steen
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods; D058954:Pancreatic Stellate Cells / Q000378:metabolism*; D050500:Pancreatitis, Chronic / Q000209:etiology; D040901:Proteomics / Q000379:methods*; D051381:Rats; D012269:Ribosomal Proteins / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000199:Actins; D012269:Ribosomal Proteins; C498862:Acta2 protein, rat