Highly persistent polycyclic aromatic hydrocarbon-DNA adducts in mouse skin: Detection by 32P-postlabeling analysis

Abstract

A 32P-postlabeling method for carcinogen-DNA adduct analysis recently developed in our laboratory was applied to skin DNA from mice treated topically with polycyclic aromatic hydrocarbons (PAHs). After application of 4 doses of 1.2 μmol each of benzo[ a]pyrene (BP), 3-methylcholanthrene (MC) and 7,12-dimethylbenz[ a]anthracene (DMBA), respectively, total covalent adduct binding in mouse skin DNA initially amounted to 1 adduct in 6.0 × 10 4–1.3 × 10 5 nucleotides. Four weeks after treatment, these levels had declined to 1 adduct in 1.4 × 10 6–2.7 × 10 6 nucleotides. Substantial removal of DNA adducts occurred during the first 2 weeks after carcinogen application while adducts remaining thereafter underwent little or no repair between 2 and 4 weeks after treatment. These results raise the possibility that the persistent adducts occupy specific genomic sites in quiescent cells where they may not be amenable to repair because of localized conformational alterations of DNA or shielding by associated proteins.