title: Proteomic analysis of an immortalized mouse pancreatic stellate cell line identifies differentially-expressed proteins in activated vs nonproliferating cell states.
abstract: Pancreatic stellate cells (PaSC) are mediators in chronic pancreatitis and pancreatic cancer pathogenesis. Proteins regulating the biomolecular pathways involved in the conversion of activated to quiescent PaSC may have a significant influence in the development of chronic pancreatitis. We aim to compare differentially expressed proteins from an immortalized cell line of mouse PaSC in the activated and serum-starved cell states using mass spectrometry-based proteomics. PaSC cultured in media supplemented with fetal bovine serum (FBS) proliferate in the activated state, while serum starvation promotes the cellular transition to a "pseudo-quiescent" state. Using these two cell states, we performed a comparative mass spectrometry (GeLC-MS/MS) proteomic analysis. We identified over 2000 nonredundant proteins in PaSC. Qualitative and label-free quantitative analysis revealed several hundred proteins that were differentially abundant between the cell states. Proteins that were more abundant in activated PaSC included cytoskeletal proteins and ribosomal proteins, while those more abundant in pseudoquiescent PaSC included proteins involved in protein degradation-related pathways (lysosome, ubiquitin-mediated proteolysis, and the proteasome). Investigation of the role of PaSC in the pathogenesis of chronic pancreatitis using the mass spectrometry-based proteomics strategy described herein will lead to further insights into the molecular mechanisms associated with the disease.
authors.affiliation: Center for Pancreatic Disease, Division of Gastroenterology, Hepatology and Endoscopy, Brigham and Women's Hospital and Department of Medicine, Harvard Medical School , Boston, Massachusettes, United States.
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: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D019295:Computational Biology / Q000379:methods; D003599:Cytoskeleton / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005355:Fibrosis; D005786:Gene Expression Regulation; D051379:Mice; D058954:Pancreatic Stellate Cells / Q000166:cytology*; D010195:Pancreatitis / Q000473:pathology; D040901:Proteomics / Q000379:methods*; D012269:Ribosomal Proteins / Q000737:chemistry; D013347:Subcellular Fractions / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D012269:Ribosomal Proteins