title: Genomic and gene expression responses to genotoxic stress in PAC2 zebrafish embryonic cell line.
abstract: PAC2 cell line is, along most of the developed zebrafish cell lines, poorly characterized concerning its response to genotoxicants. To define the PAC2 cell line response to different forms of genotoxic stress, we exposed the cells to model genotoxic agents (benzo[a]pyrene, B[a]P, and ethyl methanesulfonate) and subsequently monitored DNA damage and alterations by using the battery of tests, including the Comet assay, quantitative random-amplified polymorphic DNA and amplified fragment length polymorphism. The expression of several DNA repair (xpc, xpd, hr23b, rad51, msh2) and oxidative stress response (sod (Cu/Zn)) genes was monitored as well. To obtain an indication of the PAC2 cell line metabolizing capacity, the expression of genes belonging to cyp1, cyp2 and cyp3 families was assessed upon exposure to B[a]P. Genotoxic responses were observed in all the used methods, and quantitative random-amplified polymorphic DNA and amplified fragment length polymorphism proved to be more sensitive by revealing DNA alterations even when the Comet assay indicated lack of significant damage. The PAC2 cell line demonstrated basal and B[a]P-induced expression of several cyp genes, suggesting its ability to metabolize indirect acting xenobiotics to a certain point. Based on these results, PAC2 cells seem to be sensitive zebrafish in vitro model in the genotoxicity assessment of the direct acting genotoxicant; however, they are less sensitive toward the indirect acting genotoxicant due to their limited metabolizing properties.
authors.affiliation: Department of Zoology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000, Zagreb, Croatia.
authors.forename: Maja
authors.initials: M
authors.lastname: Šrut
authors.affiliation: Arcachon Marine Station, CNRS, UMR EPOC 5805, University of Bordeaux, Place du Docteur Peyneau, 33120, Arcachon, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Bourdineaud
authors.affiliation: Department of Zoology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000, Zagreb, Croatia.
authors.forename: Anamaria
authors.initials: A
authors.lastname: Štambuk
authors.affiliation: Department of Zoology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, 10000, Zagreb, Croatia.
authors.forename: Göran I V
authors.initials: GI
authors.lastname: Klobučar
mesh_terms: D054458:Amplified Fragment Length Polymorphism Analysis; D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity*; D002460:Cell Line; D020552:Comet Assay; D004249:DNA Damage / Q000187:drug effects*; D004260:DNA Repair / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D015027:Zebrafish / Q000196:embryology / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D029961:Zebrafish Proteins; D001564:Benzo(a)pyrene
keywords: AFLP; Comet assay; DNA repair genes; quantitative RAPD; xenobiotic metabolism