(7S,8S)--Dihydroxy--7,8--dihydrobenzo[a]
pyrene ((+)-BP-7,8-diol) is epoxidized to (7S,8R)-dihydroxy-(9S,10R)-epoxy-7,8,9,10-tetrahydrobenzo[a]
pyrene ((+)-syn-
BPDE) by
cytochrome P-450 isoenzymes and to (7S,8R)-dihydroxy-(9R,10S)-epoxy-7,8,9,10-tetrahydrobenzo[a]
pyrene ((-)-anti-
BPDE) by peroxyl
free radicals. 32P postlabeling analysis of the diastereomeric
BPDE-DNA adducts was used to investigate the pathways of (+)-BP-7,8-diol oxidation in mouse skin in vivo. The pattern of deoxynucleoside 3',5'-bisphosphate adducts in epidermal scrapings from female CD-1 mice indicated that
cytochrome P-450 was the major
oxidant. Similar results were obtained when the
tumor-promoting
phorbol ester tetradecanoylphorbolacetate (TPA) was coadministered with (+)-BP-7,8-diol. However, when animals were pretreated with TPA 24 h before coadministration of TPA and (+)-BP-7,8-diol, the pattern of
BPDE-DNA adducts indicated that peroxyl radicals made a major contribution to (+)-BP-7,8-diol epoxidation.
Peroxyl radical-dependent epoxidation was maximal when the time between the two TPA administrations was 24-72 h. No increase in (-)-anti-
BPDE-DNA was observed when the non-
tumor-promoting
phorbol ester 4-O-methyl-TPA was substituted for TPA. The
calcium ionophore A23187 stimulated
peroxyl radical generation when substituted for the first, but not the second, TPA treatment. The antiinflammatory
steroid fluocinolone acetonide inhibited (-)-anti-
BPDE-DNA adduct formation when coadministered with the first but not the second TPA treatment. These findings demonstrate the existence of two independent pathways of metabolic activation of (+)-BP-7,8-diol in mouse epidermis, one dependent on
cytochrome P-450 and the other dependent on peroxyl
free radicals. The results also suggest that repetitive
topical administration of
tumor-promoting
phorbol esters remodels epidermal metabolism leading to a significant increase in
free radical generation.