RELATIONSHIP BETWEEN ADDUCT FORMATION, RATES OF EXCISION-REPAIR AND THE CYTOTOXIC AND MUTAGENIC EFFECTS OF STRUCTURALLY-RELATED POLYCYCLIC AROMATIC CARCINOGENS

The cytotoxic and mutagenic effect of l-nitrosopyrene (1-NOP) and N-ac etoxy-2-acetylaminofluarene (N-AcO-AAF) were compared with that of (+/ -)-7 beta,8 alpha-dihydroxy-9 alpha,10-epoxy-7,8,9,10-tetrahydrobenzo[ a]pyrene (BPDE) as a function of the initial frequency of adducts form ed in the DNA of repair-proficient diploid human fibroblasts and the f raction remaining at the time the cells replicate their DNA. The princ ipal adducts of all three agents involve guanine. The initial level of BPDE-, 1-NOP-, or N-AcO-AAF-induced adducts per 10(6) nucleotides req uired to lower the survival of these cells to 37% of the control was 8 , 25, and 50, respectively. The frequency of mutants per 10(6) clonabl e cells induced at those levels of initial adduct formation was 160, 8 0, and 40, respectively. We determined the rate of excision repair of these adducts from the overall genome, from the individual strands of the hypoxanthine phosphoribosyltransferase (HPRT) gene, and in the cas e of 1-NOP and BPDE, at the level of individual nucleotides in the non transcribed strand of exon 3 of that gene, a region where mutations in duced by those agents are particularly frequent, 1-NOP-induced adducts were excised from the overall genome and from the individual strands of HPRT at a rate 2-3 times faster than BPDE-induced adducts. The aver age rate of repair of 1-NOP-induced adducts in exon 3 was also 2-3 tim es faster than the average rate of repair of BPDE-induced adducts. How ever, at particular nucleotides 1-NOP-induced adducts were repaired mu ch faster, or slower, or in some cases at a rate equal to that of BPDE -induced adducts. Excision repair of N-AcO-AAF-induced adducts (i.e., deacetylated aminofluorene residues) was significantly slower than tha t of BPDE- or 1-NOP-induced adducts, and was not strand-specific. In a n in vitro assay, BPDE adducts were four times more effective in block ing transcription than were 1-NOP or N-AcO-AAF-induced adducts. We con clude that the cytotoxic and mutagenic effect of these carcinogens ref lect a complex interplay of adduct conformation, ability of adducts to block replication and transcription, and variation in the rate of exc ision repair, even at the nucleotide level.