title: DIGE-based protein expression analysis of B[a]P-exposed hepatoma cells reveals a complex stress response including alterations in oxidative stress, cell cycle control, and cytoskeleton motility at toxic and subacute concentrations.
abstract: Although the effects of high concentrations of the carcinogen benzo[a]pyrene (B[a]P) have been studied extensively, little is known about its effects at subacute toxic concentrations, which are typical for environmental pollutants. We exposed murine Hepa1c1c7 cells to a toxic concentration (5 μM) and a subacute concentration (50 nM) of B[a]P over a period of 2-24 h to differentiate between acute and pseudochronic effects and conducted a time-course analysis of B[a]P-influenced protein expression by DIGE. In total, a set of 120 spots were found to be significantly altered due to B[a]P exposure of which 112 were subsequently identified by mass spectrometry. Clustering and principal component analysis were conducted to identify sets of proteins responding in a concerted manner to the exposure. Our results indicate an immediate response to the contaminant at the protein level and demonstrate that B[a]P exposure alters the cellular response by disturbing proteins involved in oxidative stress, cell cycle regulation, apoptosis, and cytoskeleton organization. Furthermore, network analysis of protein-protein interactions revealed a complex network of interacting, B[a]P-regulated proteins mostly belonging to the cytoskeleton organization and several signal transduction pathways.
authors.affiliation: Department of Proteomics, UFZ, Helmholtz-Centre for Environmental Research, Leipzig, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Dautel
authors.forename: Stefan
authors.initials: S
authors.lastname: Kalkhof
authors.forename: Saskia
authors.initials: S
authors.lastname: Trump
authors.forename: Jacob
authors.initials: J
authors.lastname: Michaelson
authors.forename: Andreas
authors.initials: A
authors.lastname: Beyer
authors.forename: Irina
authors.initials: I
authors.lastname: Lehmann
authors.forename: Martin
authors.initials: M
authors.lastname: von Bergen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001564:Benzo(a)pyrene / Q000633:toxicity*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000378:metabolism; D030562:Databases, Protein; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects*; D025941:Protein Interaction Mapping; D020543:Proteome / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D018675:Toxicity Tests
chemical_list: D020543:Proteome; D001564:Benzo(a)pyrene