title: Differential protein profiling of primary versus immortalized human RPE cells identifies expression patterns associated with cytoskeletal remodeling and cell survival.
abstract: Functional research of retinal pigment epithelium (RPE) most often relies on utilization of RPE-derived cell lines in vitro. However, no studies about similarities and differences of the respective cell lines exist so far. Thus, we here analyze the proteome of the most popular RPE cell lines: ARPE-19 and hTERT and compare their constitutive and de novo synthesized protein expression profiles to human early passage retinal pigment epithelial cells (epRPE) by 2-D electrophoresis and MALDI-TOF peptide mass fingerprinting. In all three cell lines the baseline protein expression pattern corresponded well to the de novo synthesized cellular proteome. However, comparison of the protein profile of epRPE cells with that of hTERT-RPE cells revealed a higher abundance of proteins related to cell migration, adhesion, and extracellular matrix formation, paralleled by a down-regulation of proteins attributed to cell polarization, and showed an altered expression of detoxification enzymes in hTERT-RPE. ARPE-19 cells, however, exhibited a higher abundance of components of the microtubule cytoskeleton and differences in expression of proteins related to proliferation and cell death. epRPE cells, hTERT-RPE, and ARPE-19 therefore may respond differently with respect to certain functional properties, a finding that should prove valuable for future in vitro studies.
authors.affiliation: Department of Ophthalmology, Ludwig-Maximilians-University, Munich, Germany, and GSF Research Center for Environment and Health, Neuherberg, Germany.
authors.forename: Claudia S
authors.initials: CS
authors.lastname: Alge
authors.forename: Stefanie M
authors.initials: SM
authors.lastname: Hauck
authors.forename: Siegfried G
authors.initials: SG
authors.lastname: Priglinger
authors.forename: Anselm
authors.initials: A
authors.lastname: Kampik
authors.forename: Marius
authors.initials: M
authors.lastname: Ueffing
mesh_terms: D018929:Cell Culture Techniques; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival; D002478:Cells, Cultured; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D010449:Peptide Mapping; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000378:metabolism*; D040081:Protein Array Analysis / Q000379:methods*; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014404:Tubulin / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D020543:Proteome; D014404:Tubulin