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- W1988387694 abstract "The formation of DNA adducts was investigated in mouse fibroblasts treated with dibenzo[a,e]fluoranthene (DBF), using the nuclease P1 modification of the 32P-post-labeling method. In order to separate the poorly soluble, bulky DNA adducts of this potent sarcomogenic, six-ring polycyclic aromatic hydrocarbon, several modifications of the method were introduced. Chromatographic spots were identified by incubating fibroblasts with the four major proximate metabolites of DBF and observing the co-migration of adducts with those of DBF. DNA-DBF adducts chromatographed very reproducibly in three major spots and in greater than 10 spots of medium or low importance. The most prominent spots, 2 and 3, were present characteristically after incubation of cells with the DBF-bay region dihydrodiol (+/- -trans-3,4-dihydro-3,4-dihydroxyDBF; DBF-3,4-DHD). Incubation with the DBF pseudo-bay region dihydrodiol (+/- -trans-12,13-dihydro-12,13-dihydroxyDBF; DBF-12,13-DHD) gave rise to a more complex pattern of nine spots, two of which, spots 4 and 5, were prominent. Direct in vitro reaction between DNA and the synthetic anti-isomer of the DBF-bay region DHD epoxide yielded adducts in spots 2 and 3, while the DBF-anti-pseudo-bay region DHD epoxide yielded adducts in spots 4 and 5. Peripheral, fast-migrating spots present in the DBF chromatogram were identified as adducts of DBF-7OH-3,4-DHD and DBF-3OH-12,13-DHD. Major spot 1 was present in all DBF chromatograms but not after incubation with the DBF bay and pseudo-bay region proximate metabolites. Its probable origin as a non-bay region epoxide reaction is discussed. In previous experiments, the physicochemically very similar DBF-bay region and pseudo-bay region tritium-labeled adducts co-eluted in HPLC as a single peak. 32P-Post-labeling analysis allowed reproducible separation of DBF-DNA adducts and showed in addition the existence of several new adducts models of DBF. Quantification of DBF adducts made it possible to identify the DBF-bay region DHD epoxide and the metabolites responsible for spot 1 adducts as the major ultimate DBF metabolites in fibroblasts." @default.
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- W1988387694 date "1990-01-01" @default.
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- W1988387694 title "32P-Post-labelling analysis of DNA adducts in mouse embryo fibroblasts treated with dibenzo[a,e]fluoranthene and its major metabolites" @default.
- W1988387694 doi "https://doi.org/10.1093/carcin/11.2.301" @default.
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