Genotoxicity and induction of DNA damage responsive genes by food-borne heterocyclic aromatic amines in human hepatoma HepG2 cells.

Abstract

Heterocyclic aromatic amines (HAAs) are potential human carcinogens formed in well-done meats and fish. The most abundant are 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-Amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline (4,8-DiMeIQx) and 2-Amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ). HAAs exert genotoxic activity after metabolic transformation by CYP1A enzymes, that is well characterized, however the genomic and intervening responses are not well explored. We have examined cellular and genomic responses of human hepatoma HepG2 cells after 24h exposure to HAAs. Comet assay revealed increase in formation of DNA strand breaks by PhIP, MeIQx and IQ but not 4,8-DiMeIQx, whereas increased formation of micronuclei was not observed. The four HAAs up-regulated expression of genes encoding metabolic enzymes CYP1A1, CYP1A2 and UGT1A1 and expression of TP53 and its downstream regulated genes CDKN1A, GADD45α and BAX. Consistent with the up-regulation of CDKN1A and GADD45α the cell-cycle analysis showed arrest in S-phase by PhIP and IQ, and in G1-phase by 4,8-DiMeIQx and MeIQx. The results indicate that upon exposure to HAAs the cells respond with the cell-cycle arrest, which enables cells to repair the damage or eliminate them by apoptosis. However, elevated expression of BCL2 and down-regulation of BAX may indicate that HAAs could suppress apoptosis meaning higher probability of damaged cells to survive and mutate.

• Authors: Marko Pezdirc, Bojana Žegura, Metka Filipič
• Journal: Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association (Food Chem Toxicol) Vol. 59 Pg. 386-94 (Sep 2013) ISSN: 1873-6351 [Electronic] England
• PMID: 23810796 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier Ltd. All rights reserved.

Chemical References

• 2-amino-3-methyl-3H-imidazo(4,5-f)quinoline
• Carcinogens
• Imidazoles
• Mutagens
• Quinolines
• Quinoxalines
• 2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline
• 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine
• 3,4,8-trimethylimidazo(4,5-f)quinoxalin-2-amine

Topics

• Biotransformation
• Carcinogens (metabolism, toxicity)
• Cell Cycle (drug effects)
• Cell Survival (drug effects)
• DNA Breaks (drug effects)
• DNA Damage
• Food Contamination
• Gene Expression Regulation (drug effects)
• Hep G2 Cells
• Hepatocytes (cytology, drug effects, enzymology, metabolism)
• Hot Temperature (adverse effects)
• Humans
• Imidazoles (metabolism, toxicity)
• Mutagens (metabolism, toxicity)
• Quinolines (metabolism, toxicity)
• Quinoxalines (metabolism, toxicity)