Benzo[ a]pyrenediol-epoxide induces loss of heterozygosity in Chinese hamster ovary cells heterozygous at the aprt locus

Abstract

Benzo[ a]pyrenediol-epoxide (BPDE), a metabolite of the ubiquitous environmental carcinogen benzo[ a]pyrene (B[ a]P), has been implicated as a point mutagen. However, as mutational events other than point mutations are also often associated with cancer, we have investigated whether BPDE can induce other classes of mutation. This was done by analyzing mutation at the aprt and hprt loci, both in hemizygous (D422) and heterozygous (D423) Chinese hamster ovary (CHO) cell strains. Southern blotting analysis indicated that BPDE is not an effective producer of either delections or insertions in the hemizygous environment. The analysis of mutation in the aprt heterozygote was done to investigate the frequency of loss of heterozygosity (LOH) events following BPDE treatment. Using PCR to produce an artificial restriction fragment length polymorphism in the functional aprt allele, BPDE was found to induce LOH in about one-quarter of the mutants recovered. While the precise mechanism of this phenomenon remains obscure, it is likely to have important implications, since similar events involving homologous recombination in somatic cells may have an impact in tumorigenesis.