Journal Article

Immunohistochemical detection of 1,<i>N</i><sup>6</sup>-ethenodeoxyadenosine, a promutagenic DNA adduct, in liver of rats exposed to vinyl chloride or an iron overload

Yan Yang, Jagadeesan Nair, Alain Barbin and Helmut Bartsch

in Carcinogenesis

Volume 21, issue 4, pages 777-781
Published in print April 2000 | ISSN: 0143-3334
Published online April 2000 | e-ISSN: 1460-2180 | DOI:
Immunohistochemical detection of 1,N6-ethenodeoxyadenosine, a promutagenic DNA adduct, in liver of rats exposed to vinyl chloride or an iron overload

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Etheno adducts in DNA bases are formed from exogenous agents such as vinyl chloride and urethane, but also via endogenous lipid peroxidation products like trans-4-hydroxy-2-nonenal. An immunohistochemical method was developed to localize the promutagenic 1,N6-ethenodeoxyadenosine DNA adduct in liver of rats exposed to vinyl chloride or an iron overload with or without carbon tetrachloride. Six monoclonal antibodies, previously produced through collaborative efforts, were screened for their optimal adduct recognition and low background formation. The antibody generated by clone EM-A-4 was found to be most suitable. Semi-quantitative image analysis of relative pixel intensity showed ~1.5 times higher adduct levels (P < 0.05) in the livers of rats treated with vinyl chloride or an iron overload when compared with untreated controls. Significantly elevated adduct levels persisted in vinyl chloride-treated rat liver 14 days after cessation of exposure, suggesting that this adduct is not rapidly eliminated from rat liver DNA. Using the new immunohistochemical method it is possible to visualize this promutagenic etheno-DNA adduct that may play a role in oxidative stress and lipid peroxidation-induced DNA damage in carcinogenesis.

Keywords: DAB, 3,3′-diaminobenzidine; εdA, 1,N6-ethenodeoxyadenosine; εdC, 3,N4-ethenodeoxycytidine; LPO, lipid peroxidation; PBS, phosphate-buffered saline; PBST, 0.2% Triton-X100 in PBS; RPI, relative pixel intensity; VC, vinyl chloride.

Journal Article.  2984 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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