Alcohol Metabolism in Human Cells Causes DNA Damage and Activates the Fanconi Anemia–Breast Cancer Susceptibility (FA-BRCA) DNA Damage Response Network

Authors

  • Jessy Abraham,

    1. From the Section on Molecular Neurobiology (JA, PJB), Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, NIH, Rockville, Maryland; Masonic Cancer Center, University of Minnesota (SB), Minneapolis, Minnesota; and Division of Gastroenterology and Hepatology, Department of Medicine (DC), Indiana University School of Medicine and Roudebush Veterans Administration Medical Center, Indianapolis, Indiana.
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  • Silvia Balbo,

    1. From the Section on Molecular Neurobiology (JA, PJB), Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, NIH, Rockville, Maryland; Masonic Cancer Center, University of Minnesota (SB), Minneapolis, Minnesota; and Division of Gastroenterology and Hepatology, Department of Medicine (DC), Indiana University School of Medicine and Roudebush Veterans Administration Medical Center, Indianapolis, Indiana.
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  • David Crabb,

    1. From the Section on Molecular Neurobiology (JA, PJB), Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, NIH, Rockville, Maryland; Masonic Cancer Center, University of Minnesota (SB), Minneapolis, Minnesota; and Division of Gastroenterology and Hepatology, Department of Medicine (DC), Indiana University School of Medicine and Roudebush Veterans Administration Medical Center, Indianapolis, Indiana.
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  • Phillip J. Brooks

    1. From the Section on Molecular Neurobiology (JA, PJB), Laboratory of Neurogenetics, National Institute on Alcohol Abuse and Alcoholism, NIH, Rockville, Maryland; Masonic Cancer Center, University of Minnesota (SB), Minneapolis, Minnesota; and Division of Gastroenterology and Hepatology, Department of Medicine (DC), Indiana University School of Medicine and Roudebush Veterans Administration Medical Center, Indianapolis, Indiana.
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Reprint requests: Phillip J. Brooks, National Institute on Alcohol Abuse and Alcoholism, Division of Metabolism and Health Effects, 5635 Fishers Lane, MSC 9304, Bethesda, MD 20892-9304; Tel.: 301-402-0883; Fax: 301-594-0673; E-mail: pjbrooks@mail.nih.gov

Abstract

Background:  We recently reported that exposure of human cells in vitro to acetaldehyde resulted in the activation of the Fanconi anemia–breast cancer susceptibility (FA-BRCA) DNA damage response network.

Methods:  To determine whether intracellular generation of acetaldehyde from ethanol metabolism can cause DNA damage and activate the FA-BRCA network, we engineered HeLa cells to metabolize alcohol by expression of human alcohol dehydrogenase (ADH) 1B.

Results:  Incubation of HeLa-ADH1B cells with ethanol (20 mM) resulted in acetaldehyde accumulation in the media, which was prevented by co-incubation with 4-methyl pyrazole (4-MP), a specific inhibitor of ADH. Ethanol treatment of HeLa-ADH1B cells produced a 4-fold increase in the acetaldehyde–DNA adduct and N2-ethylidene-dGuo and also resulted in the activation of the FA-BRCA DNA damage response network, as indicated by a monoubiquitination of FANCD2 and phosphorylation of BRCA1. Ser 1524 was identified as 1 site of BRCA1 phosphorylation. The increased levels of DNA adducts, FANCD2 monoubiquitination, and BRCA1 phosphorylation were all blocked by 4-MP, indicating that acetaldehyde, rather than ethanol itself, was responsible for all 3 responses. Importantly, the ethanol concentration we used is within the range that can be attained in the human body during social drinking.

Conclusions:  Our results indicate that intracellular metabolism of ethanol to acetaldehyde results in DNA damage, which activates the FA-BRCA DNA damage response network.

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