title: Increased cytotoxicity of food-borne mycotoxins toward human cell lines in vitro via enhanced cytochrome p450 expression using the MTT bioassay.
abstract: Eight food-borne mycotoxins epidemiologically implicated in human disease were tested for their cytotoxic effects on human cells previously immortalised and transfected to introduce human cytochrome p450 (CYP 450) genes. Such cells retain many characteristics of normal cell growth and differentiation while simultaneously having the potential of either increasing or decreasing the metabolic activity (cytotoxicity) of the challenging mycotoxins. The MTT assay provided an indication of cytotoxicity. Of the nine CYP450s introduced CYP1A2 was most effective, rendering the cells 540 times more sensitive than the control cells to aflatoxin B1, 28 times more sensitive to aflatoxin G1 and 8-fold more sensitive to ochratoxin A. CYP3A4 resulted in the cells being 211 times more toxic to aflatoxin B1 and 8-fold more toxic to aflatoxin G1 while CYP 2A6, CYP 3A5 and CYP 2E1 also produced observable effects. No increase in metabolic activity was found using cyclopiazonic acid, deoxynivalenol, fumonisin B1, patulin or T-2 toxin. CD5Os were calculated for the mycotoxins against the non-CYP-introduced control cells. There was almost a five order of magnitude difference between the most toxic, T-2 toxin (CD50 0.0057 microgram/ml) and the least toxic, fumonisin B1 (CD50 476.2 micrograms/ml). In vitro biological assays thus provide an excellent system for quantifying the often low CD50s expressed by mycotoxins in foods.
authors.affiliation: Department of Bioscience and Biotechnology, University of Strathclyde, 204 George Street, Glasgow G1 1XW, UK. cwlewis@clara.co.uk
authors.forename: C W
authors.initials: CW
authors.lastname: Lewis
authors.forename: J E
authors.initials: JE
authors.lastname: Smith
authors.forename: J G
authors.initials: JG
authors.lastname: Anderson
authors.forename: R I
authors.initials: RI
authors.lastname: Freshney
mesh_terms: D001681:Biological Assay / Q000379:methods; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005506:Food Contamination; D006801:Humans; D009183:Mycotoxins / Q000494:pharmacology / Q000633:toxicity*; D013778:Tetrazolium Salts / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism*
chemical_list: D009183:Mycotoxins; D013778:Tetrazolium Salts; D013844:Thiazoles; D003577:Cytochrome P-450 Enzyme System; C022616:thiazolyl blue