title: Use of human liver S9 in the Ames test: assay of three procarcinogens using human S9 derived from multiple donors.
abstract: The purpose of the present study was to examine the inter-individual variation in the mutagenicity of chemicals using a variety of human S9 fractions. For this purpose, three procarcinogens, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), benzo[a]pyrene (BP), and dimethylnitrosamine (DMN), were selected for the Ames test and their mutagenicity was examined using human liver S9 fractions prepared from 18 different donors and one pooled liver S9 fraction prepared from 15 different donors. In addition, rat S9 fraction prepared from male rats pretreated with phenobarbital and 5,6-benzoflavone (PB/BF) was used as reference in order to examine the mutagenic differences between human and rat (PB/BF) S9 fractions. The data demonstrate a large inter-individual diversity in the mutagenic response to procarcinogens. The mutagenicity of IQ and BP in the presence of a human liver S9 fraction (lot HLS-014) was equal to that observed in the presence of rat (PB/BF) S9 fraction. The mutagenicity of IQ and BP in the presence of a pooled human liver S9 fraction was lower (90 and 95%, respectively) than that observed in the presence of rat (PB/BF) S9. On the contrary, the mutagenicity of DMN in the presence of either a selected human liver S9 fraction (lot HLS-014) or pooled fraction was 8-fold higher than that found in the presence of rat (PB/BF) S9 fraction. Human liver S9 fraction (lot HLS-014) had one of the highest cytochrome P450 enzyme activities among the 18 different donors and higher than the pooled human liver S9 fraction. These results suggest that the use of both selected human liver S9 fractions with high metabolic activity (e.g., lot HLS-014 as used in this study) and a pooled S9 fraction with moderate metabolic activity could be used as a means to evaluate the inter-individual variability in mutagenic response to chemicals and to confirm positive responses from studies completed with rodent S9.
authors.affiliation: Department of Developmental Safety Assessment Research, Eisai Co, Ltd, 1, Takehaya-machi, Kawashima-cho, Hashima-gun, Gifu 501-6195, Japan. a-hakura@hhc.eisai.co.jp
authors.forename: Atsushi
authors.initials: A
authors.lastname: Hakura
authors.forename: Satoshi
authors.initials: S
authors.lastname: Suzuki
authors.forename: Shigeki
authors.initials: S
authors.lastname: Sawada
authors.forename: Tadakazu
authors.initials: T
authors.lastname: Sugihara
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hori
authors.forename: Kanako
authors.initials: K
authors.lastname: Uchida
authors.forename: William D
authors.initials: WD
authors.lastname: Kerns
authors.forename: Fumio
authors.initials: F
authors.lastname: Sagami
authors.forename: Satoru
authors.initials: S
authors.lastname: Motooka
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Satoh
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D001564:Benzo(a)pyrene / Q000633:toxicity*; D002273:Carcinogens / Q000378:metabolism / Q000633:toxicity*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004128:Dimethylnitrosamine / Q000633:toxicity*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D008875:Middle Aged; D009152:Mutagenicity Tests / Q000379:methods; D011804:Quinolines / Q000633:toxicity*; D051381:Rats; D012486:Salmonella typhimurium / Q000187:drug effects / Q000235:genetics; D013045:Species Specificity
chemical_list: D002273:Carcinogens; D011804:Quinolines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; D001564:Benzo(a)pyrene; D003577:Cytochrome P-450 Enzyme System; D004128:Dimethylnitrosamine