title: Modulation of CYP3A4 activity alters the cytotoxicity of lipophilic phycotoxins in human hepatic HepaRG cells.
abstract: The aim of this study was to investigate (i) the cytotoxic effects of lipophilic phycotoxins, including okadaic acid (OA) and dinophysistoxin-1 and -2 (DTX-1 and DTX-2), pectenotoxin-2 (PTX-2), yessotoxin (YTX), spirolide (SPX), and azaspiracids-1, -2 and -3 (AZA-1, AZA-2 and AZA-3), in human HepaRG cells using a multiparametric high content analysis approach, (ii) the ability of nine lipophilic phycotoxins to act as PXR agonists in a HepG2-PXR cell line, (iii) their potential to induce CYP450 activity, and (iv) the role of CYP3A4 in cytotoxicity induced by lipophilic phycotoxins. Our results indicate that while OA, DTX-1 and DTX-2 activated PXR-dependent transcriptional activity in HepG2 cells, no increase of CYP450 (1A2, 3A4, 2C9, 2C19) activities were observed in HepaRG cell following a 72h treatment with these toxins. Multiparametric analysis showed that OA, DTX-1, DTX-2, and PTX-2 were highly cytotoxic in HepaRG cells; inducing cell loss, activation of caspase-3 and γ-H2AX formation. However, no toxicity was observed for YTX, SPX, and AZAs. Moreover, we found that inhibition of CYP3A4 activity by ketoconazole enhances the toxic effects of OA, DTX-1, DTX-2, and PTX-2 in HepaRG cells. Taken together, these results suggest that CYP3A4-mediated metabolism of some lipophilic phycotoxins decreases their in vitro toxicity.
authors.affiliation: ANSES, Fougères Laboratory, Toxicology of Contaminants Unit, French Agency for Food, Environmental and Occupational Health & Safety, Fougères 35302, France. Electronic address: pierre-jean.ferron@anses.fr.
authors.forename: P J
authors.initials: PJ
authors.lastname: Ferron
authors.affiliation: ANSES, Fougères Laboratory, Toxicology of Contaminants Unit, French Agency for Food, Environmental and Occupational Health & Safety, Fougères 35302, France.
authors.forename: K
authors.initials: K
authors.lastname: Hogeveen
authors.affiliation: INRA, UMR 1331 TOXALIM, 400 route des Chappes, BP 167, 06903, Sophia-Antipolis, France.
authors.forename: G
authors.initials: G
authors.lastname: De Sousa
authors.affiliation: INRA, UMR 1331 TOXALIM, 400 route des Chappes, BP 167, 06903, Sophia-Antipolis, France.
authors.forename: R
authors.initials: R
authors.lastname: Rahmani
authors.affiliation: ANSES, Fougères Laboratory, Analysis of Residues and Contaminants Unit, French Agency for Food, Environmental and Occupational Health & Safety, Fougères 35302, France.
authors.forename: E
authors.initials: E
authors.lastname: Dubreil
authors.affiliation: ANSES, Fougères Laboratory, Toxicology of Contaminants Unit, French Agency for Food, Environmental and Occupational Health & Safety, Fougères 35302, France.
authors.forename: V
authors.initials: V
authors.lastname: Fessard
authors.affiliation: ANSES, Fougères Laboratory, Toxicology of Contaminants Unit, French Agency for Food, Environmental and Occupational Health & Safety, Fougères 35302, France.
authors.forename: L
authors.initials: L
authors.lastname: Le Hegarat
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism*; D065692:Cytochrome P-450 CYP3A Inhibitors / Q000494:pharmacology; D004249:DNA Damage; D005663:Furans / Q000633:toxicity; D006657:Histones / Q000378:metabolism; D006801:Humans; D007654:Ketoconazole / Q000494:pharmacology; D008099:Liver / Q000166:cytology; D008387:Marine Toxins / Q000633:toxicity; D019319:Okadaic Acid / Q000633:toxicity; D035964:Oxocins / Q000633:toxicity; D011714:Pyrans / Q000633:toxicity; D013141:Spiro Compounds / Q000633:toxicity
chemical_list: D065692:Cytochrome P-450 CYP3A Inhibitors; D005663:Furans; C468783:H2AX protein, human; D006657:Histones; D008387:Marine Toxins; D035964:Oxocins; D011714:Pyrans; D013141:Spiro Compounds; D019319:Okadaic Acid; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human; D053148:Caspase 3; D007654:Ketoconazole
keywords: CYP3A4; Cytotoxicity; Hepatocytes; High content analysis; Phycotoxins