title: Photolytic degradation products of two highly brominated flame retardants cause cytotoxicity and mRNA expression alterations in chicken embryonic hepatocytes.
abstract: Tetradecabromo-1,4-diphenoxybenzene (TeDB-DiPhOBz) and 2,2',3,3',4,4',5,5',6,6'-decabromodiphenyl ether (BDE-209) are photolytically unstable flame retarding chemicals. Here, photocatalyzed byproducts of TeDB-DiPhOBz and BDE-209 (i.e Br(8)- to Br(11)-PB-DiPhOBz congeners from TeDB-DiPhOBz, and Br(6)- to Br(8)-BDE congeners from BDE-209), formed after 21 days of natural sunlight irradiation (SI), were assessed for exposure effects on cytotoxicity and mRNA expression levels of selected genes in chicken embryonic hepatocytes (CEH). CEHs were exposed for 36 h to concentrations of SI- and nonirradiated (NI)-TeDB-DiPhOBz and BDE-209. Cytotoxic effects were observed only in CEH exposed to 50 μM SI-BDE-209. Results from a custom-designed Avian ToxChip polymerase chain reaction array showed that NI-TeDB-DiPhOBz and NI-BDE-209, up to maximum concentrations of 1.9 and 9 μM, respectively, caused limited changes in mRNA levels of 27 genes from toxicologically relevant pathways, including phase I/II metabolism, the thyroid hormone pathway, lipid/cholesterol metabolism, oxidative stress, immune response, and cell death. In contrast, 12 and 14 of the 27 genes were altered after exposure to 25 μM SI-TeDB-DiPhOBz or 10 μM SI-BDE-209, respectively. Aryl hydrocarbon receptor (AhR)-related CYP1A4 mRNA levels were the most altered on the PCR array with an induction of 560- and 5200-fold after exposure to 1 or 25 μM SI-TeDB-DiPhOBz, respectively, and 2500- and 2300-fold after exposure to 1 or 10 μM SI-BDE-209, respectively. A dioxin-responsive luciferase reporter gene assay confirmed that the CYP1A4 inductions were independent of the dissolution solvents used (tetrahydrofuran/n-hexane, n-hexane, or methanol) during photolysis. Overall, degradation of TeDB-DiPhOBz and BDE-209 by natural sunlight generates byproducts that affect in vitro expression of genes, especially the AhR-mediated CYP1A4.
authors.affiliation: Ecotoxicology and Wildlife Health Division, Environment Canada, National Wildlife Research Centre, Carleton University , Ottawa, Ontario K1A 0H3, Canada.
authors.forename: Guanyong
authors.initials: G
authors.lastname: Su
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Letcher
authors.forename: Doug
authors.initials: D
authors.lastname: Crump
authors.forename: Reza
authors.initials: R
authors.lastname: Farmahin
authors.forename: John P
authors.initials: JP
authors.lastname: Giesy
authors.forename: Sean W
authors.initials: SW
authors.lastname: Kennedy
mesh_terms: D000818:Animals; D001969:Bromobenzenes / Q000378:metabolism / Q000633:toxicity*; D002478:Cells, Cultured; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D005260:Female; D005411:Flame Retardants / Q000378:metabolism / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects*; D055768:Halogenated Diphenyl Ethers / Q000378:metabolism / Q000633:toxicity*; D054879:Halogenation; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D010647:Phenyl Ethers / Q000378:metabolism / Q000633:toxicity*; D010782:Photolysis; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism; D013472:Sunlight; D018675:Toxicity Tests / Q000379:methods
chemical_list: D001969:Bromobenzenes; D005411:Flame Retardants; D055768:Halogenated Diphenyl Ethers; D010647:Phenyl Ethers; D012333:RNA, Messenger; D018336:Receptors, Aryl Hydrocarbon; C582340:tetradecabromodiphenoxybenzene; C010902:decabromobiphenyl ether