title: An in vitro investigation of endocrine disrupting effects of trichothecenes deoxynivalenol (DON), T-2 and HT-2 toxins.
abstract: Trichothecenes are a large family of chemically related mycotoxins. Deoxynivalenol (DON), T-2 and HT-2 toxins belong to this family and are produced by various species of Fusarium. The H295R steroidogenesis assay, regulation of steroidogenic gene expression and reporter gene assays (RGAs) for the detection of androgen, estrogen, progestagen and glucocorticoid (ant)agonist responses, have been used to assess the endocrine disrupting activity of DON, T-2 and HT-2 toxins. H295R cells were used as a model for steroidogenesis and gene expression studies and exposed with either DON (0.1-1000ng/ml), T-2 toxin (0.0005-5ng/ml) or HT-2 toxin (0.005-50ng/ml) for 48h. We observed a reduction in hormone levels in media of exposed cells following radioimmunoassay. Cell viability was determined by four colorimetric assays and we observed reduced cell viability with increasing toxin concentrations partly explaining the significant reduction in hormone levels at the highest toxin concentration of all three trichothecenes. Thirteen of the 16 steroidogenic genes analyzed by quantitative real time PCR (RT-qPCR) were significantly regulated (P<0.05) by DON (100ng/ml), T-2 toxin (0.5ng/ml) and HT-2 toxin (5ng/ml) compared to the control, with reference genes (B2M, ATP5B and ACTB). Whereas HMGR and CYP19 were down-regulated, CYP1A1 and CYP21 were up-regulated by all three trichothecenes. DON further up-regulated CYP17, HSD3B2, CYP11B2 and CYP11B1 and down-regulated NR5A1. T-2 toxin caused down-regulation of NR0B1 and NR5A1 whereas HT-2 toxin induced up-regulation of EPHX and HSD17B1 and down-regulation of CYP11A and CYP17. The expressions of MC2R, StAR and HSD17B4 genes were not significantly affected by any of the trichothecenes in the present study. Although the results indicate that there is no evidence to suggest that DON, T-2 and HT-2 toxins directly interact with the steroid hormone receptors to cause endocrine disruption, the present findings indicate that exposure to DON, T-2 toxin and HT-2 toxin have effects on cell viability, steroidogenesis and alteration in gene expression indicating their potential as endocrine disruptors.
authors.affiliation: Norwegian School of Veterinary Science, Oslo, Norway; Sokoine University of Agriculture, Morogoro, Tanzania. doreen.ndossi@nvh.no
authors.forename: D G
authors.initials: DG
authors.lastname: Ndossi
authors.forename: C
authors.initials: C
authors.lastname: Frizzell
authors.forename: N H
authors.initials: NH
authors.lastname: Tremoen
authors.forename: C K
authors.initials: CK
authors.lastname: Fæste
authors.forename: S
authors.initials: S
authors.lastname: Verhaegen
authors.forename: E
authors.initials: E
authors.lastname: Dahl
authors.forename: G S
authors.initials: GS
authors.lastname: Eriksen
authors.forename: M
authors.initials: M
authors.lastname: Sørlie
authors.forename: L
authors.initials: L
authors.lastname: Connolly
authors.forename: E
authors.initials: E
authors.lastname: Ropstad
mesh_terms: D018268:Adrenocortical Carcinoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism; D052244:Endocrine Disruptors / Q000633:toxicity*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D006727:Hormone Antagonists / Q000633:toxicity*; D006728:Hormones / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D042361:Mammary Glands, Human / Q000166:cytology / Q000378:metabolism; D011987:Receptors, Steroid / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D013605:T-2 Toxin / Q000031:analogs & derivatives / Q000633:toxicity; D014162:Transfection; D014255:Trichothecenes / Q000633:toxicity*
chemical_list: D017077:Culture Media, Conditioned; D052244:Endocrine Disruptors; D006727:Hormone Antagonists; D006728:Hormones; D011987:Receptors, Steroid; D014255:Trichothecenes; D008156:Luciferases; D013605:T-2 Toxin; C007262:deoxynivalenol; C012351:HT-2 toxin