Ethanol enhanced the genotoxicity of acrylamide in human, metabolically competent HepG2 cells by CYP2E1 induction and glutathione depletion

Evelyn Lamy, Yvonne Völkel, Peter H. Roos, Fekadu Kassie, Volker Mersch-Sundermann

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

In the present study, the genotoxicity of acrylamide (AA) was investigated in HepG2 cells using SCGE. Additionally, the influence of ethanol on the modulation of AA-induced DNA-migration caused by CYP2E1-upregulation and/or GSH-depletion was examined in the same cell line. For the ethanol/AA combination assays, the cells were treated with ethanol for 24 h prior to exposure to 5 mM AA for another 24 h. 1.25 to 10 mM AA-induced DNA migration (OTM) in HepG2 cells in a concentration-dependent manner, e.g., exposure to 10 mM AA, resulted in an 8-fold increase of DNA migration compared to the negative control. Treatment with 120 mM ethanol prior to exposure to 5 mM AA increased the level of DNA migration more than 2-fold as compared to cells treated with 5 mM AA alone. Immunoblotting showed a clear ethanol-induced increase of CYP2E1, which plays a pivotal role in AA toxification. Additionally, intracellular GSH levels were significantly reduced after ethanol or AA treatment. In the ethanol/AA combination experiments, GSH depletion was comparable to the additive effect of the single compounds. No induction of apoptosis (ssDNA assay), but necrosis was identified as responsible for the reduction of viability with increasing compound concentration. The data clearly show a higher genotoxic potential of ethanol/AA combination treatment compared to AA treatment alone. In conclusion, both the ethanol-mediated induction of CYP2E1 and the depletion of GSH provide a mechanistic explanation for the over-additive effects of ethanol and AA. Even though the concentrations used in this study were rather high, consequences for the dietary intake of AA-containing food and alcoholic beverages should be discussed.

Original languageEnglish (US)
Pages (from-to)74-81
Number of pages8
JournalInternational Journal of Hygiene and Environmental Health
Volume211
Issue number1-2
DOIs
StatePublished - Mar 12 2008

Bibliographical note

Funding Information:
Parts of this study were supported by the European Community grant QLK1-CT-1999-0810 for V.M.-S.

Copyright:
Copyright 2015 Elsevier B.V., All rights reserved.

Keywords

  • Acrylamide
  • CYP2E1
  • Comet assay
  • Glutathione depletion
  • HepG2
  • SCGE
  • ssDNA apoptosis assay

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