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Detection of O⁶-Methyldeoxyguanosine in Human Placental DNA^{1, 2,}

Division of Chemical Carcinogenesis, Naylor Dana Institute for Disease Prevention, American Health Foundation, Valhalla, New York 10595 [P. G. F., L. M. M., S. A. A., S. S. H.]; and Epidemiology Branch, National Institute of Environmental Health Sciences, and United States Environmental Protection Agency, Research Triangle Park, North Carolina 27711 [R. B. E.]

A monoclonal antibody specific for O⁶-methyldeoxyguanosine (O⁶-MedGuo) was developed. When used in a competitive enzyme-linked immunosorbent assay, 50% inhibition of binding was achieved with 0.51 pmol O⁶-MedGuo. When the competitive enzyme-linked immunosorbent assay was coupled with high-performance liquid chromatography, 2 mg of DNA could be analyzed giving a lower limit of detection of 0.5 µmol O⁶-MedGuo/mol deoxyguanosine. This assay was used to test for O⁶-MedGuo in DNA from placentas of smoking and nonsmoking women. Two of 10 DNA samples from smoking women and three of 10 from nonsmoking women had detectable concentrations of O⁶-MedGuo. Concentrations ranged from 0.6 to 1.6 µmol O⁶-MedGuo/mol deoxyguanosine. Activity of O⁶-alkylguanine DNA alkyltransferase was also measured. There was no apparent relationship between O⁶-alkylguanine DNA alkyltransferase activity and O⁶-MedGuo concentrations in the 20 subjects, nor did mean O⁶-alkylguanine DNA alkyltransferase activity differ between the two groups. Although no apparent relationship between smoking history and O⁶-MedGuo concentration was found in this preliminary study, this is the first report of a structurally identified DNA adduct in human placenta.

¹ This study was supported by National Cancer Institute Grant CA-32391.

² The research described in this article has been reviewed by the Health Effects Research Laboratory, U. S. Environmental Protection Agency and approved for publication. Approval does not signify that the contents necessarily reflect the views and policies of the Agency nor does mention of trade names or commercial products constitute endorsement or recommendation for use.

³ To whom requests for reprints should be addressed, at the Immunochemistry Section, Naylor Dana Institute, American Health Foundation, Dana Road, Valhalla, NY 10595.

Received 2/ 2/88. Revised 4/21/88. Accepted 5/ 4/88.

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