title: Genotoxic effects of heterocyclic aromatic amines in human derived hepatoma (HepG2) cells.
abstract: In order to study the mutagenic effects of heterocyclic aromatic amines (HAAs) in cells of human origin, five compounds, namely 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ), 2-amino-3, 4-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3, 8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx), the pyridoimidazo derivative 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), were tested in micronucleus (MN) assays with a human derived hepatoma (HepG2) cell line. All HAAs caused significant, dose-dependent effects. The activities of IQ, MeIQ, MeIQx and PhIP were similar (lowest effective concentrations 25-50 microM), whereas Trp-P-1 was effective at a dose of >/=2.1 microM. In addition, the HAAs were tested in MN assays with Chinese hamster ovary (CHO) cells and in Salmonella strain YG1024 using HepG2 cell homogenates as an activation mix. In the CHO experiments, positive results were obtained with Trp-P-1 and PhIP, whereas the other compounds were devoid of activity under all experimental conditions. The discrepancy in the responsivity of the two cell lines is probably due to differences in their acetylation capacity: enzyme measurements with 2-aminofluorene as a substrate revealed that the cytosolic acetyltransferase activity in the HepG2 cells is approximately 40-fold higher than that of the CHO cells. In the bacterial assays all five HAAs gave positive results but the ranking order was completely different from that seen in the HepG2/MN experiments (IQ > MeIQ > Trp-P-1 >/= MeIQx >> PhIP) and the mutagenic potencies of the various compounds varied over several orders of magnitude. The order obtained in bacterial tests with rat liver S9 mix was more or less identical to that seen in the tests with HepG2 cell homogenates but the concentrations of the amines required to give positive results were in general substantially lower (10(-5)-10(-1) microM). Overall, the results of the present study indicate that MN/HepG2 tests might reflect the mutagenic effects of HAAs more adequately than other in vitro mammalian cell systems due to the presence of enzymes involved in the metabolic conversion of the amines.
authors.affiliation: Institute of Cancer Research, University of Vienna, Borschkegasse 8a, A-1090 Vienna, Austria. siegfried.knasmueller@univie.ac.at
authors.forename: S
authors.initials: S
authors.lastname: Knasmüller
authors.forename: C E
authors.initials: CE
authors.lastname: Schwab
authors.forename: S J
authors.initials: SJ
authors.lastname: Land
authors.forename: C Y
authors.initials: CY
authors.lastname: Wang
authors.forename: R
authors.initials: R
authors.lastname: Sanyal
authors.forename: M
authors.initials: M
authors.lastname: Kundi
authors.forename: W
authors.initials: W
authors.lastname: Parzefall
authors.forename: F
authors.initials: F
authors.lastname: Darroudi
mesh_terms: D000588:Amines / Q000633:toxicity*; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000378:metabolism; D016466:CHO Cells / Q000187:drug effects / Q000201:enzymology; D002243:Carbolines / Q000633:toxicity; D002273:Carcinogens / Q000633:toxicity; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D006224:Cricetinae; D006571:Heterocyclic Compounds / Q000633:toxicity*; D006801:Humans; D007093:Imidazoles / Q000633:toxicity; D008113:Liver Neoplasms / Q000201:enzymology / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D011804:Quinolines / Q000633:toxicity; D011810:Quinoxalines / Q000633:toxicity; D051381:Rats; D012475:Salmonella / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D014407:Tumor Cells, Cultured
chemical_list: D000588:Amines; D002243:Carbolines; D002273:Carcinogens; D006571:Heterocyclic Compounds; D007093:Imidazoles; D009153:Mutagens; D011804:Quinolines; D011810:Quinoxalines; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; C036990:2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D001191:Arylamine N-Acetyltransferase; C036989:2-amino-3,4-dimethylimidazo(4,5-f)quinoline; C019132:3-amino-1,4-dimethyl-5H-pyrido(4,3-b)indole