title: Assessment of the genotoxic potential of indirect chemical mutagens in HepaRG cells by the comet and the cytokinesis-block micronucleus assays.
abstract: Many chemical carcinogens require metabolic activation to form genotoxic compounds in human. Standard in vitro genotoxicity assays performed with activation systems, such as rat liver S9, are recognised to lead to a high number of false positives. The aim of this study was to evaluate the suitability of differentiated human hepatoma HepaRG cells as an in vitro model system for the detection of DNA damage induced by promutagens using the comet and the cytokinesis-block micronucleus assays. Several promutagens were tested, including aflatoxin B1 (AFB1), benzo[a]pyrene (B[a]P), acrylamide, N-nitrosodimethylamine (NDMA), cyclophosphamide (CPA), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). Cytotoxicity of these compounds was assessed by measuring lactate dehydrogenase leakage. A 24 h exposure was generally needed to obtain an obvious positive response in differentiated HepaRG cells in the comet and in the cytokinesis-block micronucleus assays. Comet formation was observed with all compounds except IQ. B[a]P, CPA and AFB1 showed a dose-dependent increase in micronucleated cells, whereas no increase was observed with PhIP, IQ and acrylamide. These preliminary data on genotoxicity in differentiated HepaRG cells are promising but more chemicals must be tested to determine the ability of HepaRG cells to assess genotoxicity of chemicals in humans.
authors.affiliation: ANSES, Unité de toxicologie des contaminants, BP 90203, 35302 Fougères Cedex, France. l.lehegarat@afssa.fr
authors.forename: Ludovic
authors.initials: L
authors.lastname: Le Hegarat
authors.forename: Julie
authors.initials: J
authors.lastname: Dumont
authors.forename: Rozenn
authors.initials: R
authors.lastname: Josse
authors.forename: Sylvie
authors.initials: S
authors.lastname: Huet
authors.forename: Rachelle
authors.initials: R
authors.lastname: Lanceleur
authors.forename: Annick
authors.initials: A
authors.lastname: Mourot
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Poul
authors.forename: Christiane
authors.initials: C
authors.lastname: Guguen-Guillouzo
authors.forename: André
authors.initials: A
authors.lastname: Guillouzo
authors.forename: Valérie
authors.initials: V
authors.lastname: Fessard
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D020552:Comet Assay / Q000379:methods*; D048749:Cytokinesis / Q000187:drug effects*; D004249:DNA Damage; D004128:Dimethylnitrosamine / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology*; D015162:Micronucleus Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*; D051381:Rats
chemical_list: D009153:Mutagens; D004128:Dimethylnitrosamine