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 1-nitrosopyrene (1-NOP) and N-acetoxy-2-acetylaminofluorene (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 formed in the DNA of repair-proficient diploid human fibroblasts and the fraction remaining at the time the cells replicate their DNA. The principal adducts of all three agents involve guanine. The initial level of BPDE-, 1-NOP-, or N-AcO-AAF-induced adducts per 10(6) nucleotides required 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) clonable cells induced at those levels of initial adduct formation was 160, 80, 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 case of 1-NOP and BPDE, at the level of individual nucleotides in the nontranscribed strand of exon 3 of that gene, a region where mutations induced 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 average rate of repair of 1-NOP-induced adducts in exon 3 was also 2-3 times faster than the average rate of repair of BPDE-induced adducts. However, at particular nucleotides 1-NOP-induced adducts were repaired much 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 that of BPDE- or 1-NOP-induced adducts, and was not strand-specific. In an in vitro assay, BPDE adducts were four times more effective in blocking transcription than were 1-NOP or N-AcO-AAF-induced adducts. We conclude that the cytotoxic and mutagenic effect of these carcinogens reflect a complex interplay of adduct conformation, ability of adducts to block replication and transcription, and variation in the rate of excision repair, even at the nucleotide level.
McGregor
W G
WG
Department of Microbiology, Michigan State University, East Lansing 48824-1316, USA. mcgrego3@com.msu.edu
Wei
D
D
Chen
R H
RH
Maher
V M
VM
McCormick
J J
JJ
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000378:metabolism / Q000494:pharmacology; D000099:Acetoxyacetylaminofluorene / Q000378:metabolism / Q000494:pharmacology; D002273:Carcinogens / Q000737:chemistry*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018736:DNA Adducts / Q000096:biosynthesis*; D004260:DNA Repair; D015870:Gene Expression / Q000187:drug effects; D005796:Genes; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008297:Male; D009152:Mutagenicity Tests; D009153:Mutagens / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D011721:Pyrenes / Q000378:metabolism / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D002273:Carcinogens; D018736:DNA Adducts; D009153:Mutagens; D011721:Pyrenes; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D000099:Acetoxyacetylaminofluorene; C044707:1-nitrosopyrene; D007041:Hypoxanthine Phosphoribosyltransferase