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Chlorambucil-induced structural changes in the gpt gene of AS52 cells

Author:
Honma, M  Hayashi, M  Hackman, P  Sofuni, T  


Journal:
MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS


Issue Date:
1997


Abstract(summary):

The bifunctional alkylating agent chlorambucil (CBC) is a chemotherapeutic agent that induces a high yield of mouse germ-line mutations, apparently due to multi-locus deletions or other chromosomal rearrangements. We investigated the mutagenicity of CBC in cultured mammalian cells by comparing its effect in the AS52/gpt and CHO/hprt gene mutation assays, which detect large and small effects, respectively. CBC significantly increased the mutant frequency in the AS52/gpt assay, but not in the CHO/hprt assay, while the cytotoxic responses to CBC were similar in the two cell lines. This indicates that CBC induced predominantly large deletions or other gross structural changes, and not point or other small mutations. The mutational responses to CBC were similar to the responses to mitomycin C comparing them based on the cytotoxic responses. Molecular analysis of the gpt gene in AS52 mutant cells by electrophoresis following PCR amplification revealed that 81% of CBC-induced mutants lost the entire gpt gene, which is caused by large deletions or interchromosomal recombinations. The loss frequency was lower in spontaneous mutants (42%) and ethylmethanesulfonate-induced mutants (29%). This supports cytogenetic data showing that CBC is a potent clastogen in cultured mammalian cells, inducing predominantly large deletions and/or other gross structural alterations.


Page:
199---205


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