Genotoxic effects of heterocyclic aromatic amines in human derived hepatoma (HepG2) cells

• Published in: Mutagenesis Vol. 14; no. 6; pp. 533 - 540
• Main Authors: Knasmüller, Siegfried, Schwab, Christina E., Land, Susan J., Wang, Ching Y., Sanyal, Ratna, Kundi, Michael, Parzefall, Wolfram, Darroudi, Firouz
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01-11-1999; Oxford Publishing Limited (England)
• Subjects: acetyltransferase; Amines - toxicity; Animals; Arylamine N-Acetyltransferase - metabolism; Biological and medical sciences; Carbolines - toxicity; Carcinogens - toxicity; Carcinoma, Hepatocellular - enzymology; Carcinoma, Hepatocellular - metabolism; Cell Line; Cell Survival; Chemical mutagenesis; CHO Cells - drug effects; CHO Cells - enzymology; Cricetinae; HepG2 cells; heterocyclic amines; Heterocyclic Compounds - toxicity; Humans; Imidazoles - toxicity; Liver Neoplasms - enzymology; Liver Neoplasms - metabolism; Medical sciences; Mutagenicity Tests; Mutagens - toxicity; Quinolines - toxicity; Quinoxalines - toxicity; Rats; Salmonella; Salmonella - drug effects; Salmonella - genetics; Salmonella - growth & development; Toxicology; Tumor Cells, Cultured; Chromosomal aberration; Human; Carcinoma; Micronucleus test; Nitrogen heterocycle; Mutagenicity testing; Toxicity; Hepatic disease; Hepatocellular carcinoma; Malignant tumor; Dose activity relation; Amine; Cell line; Digestive diseases; Aromatic compound; Arylamine
• ISSN: 0267-8357; 1464-3804
• DOI: 10.1093/mutage/14.6.533

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 μM), whereas Trp-P-1 was effective at a dose of ≥2.1 μM. 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 ~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 μM). 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.