While a significant fraction of the 70,000 chemicals in commercial use in the United States are probably mutagens and/or carcinogens, only about 3,000 have been adequately tested in at least one assay. Since some of these chemicals represent a significant environmental health risk to man in terms of increased cancer and genetic disease, it is important that they be identified and characterized with the ultimate aim of limiting human exposure. To this end, many distinct assays have been developed, but each has certain inherent limitations. We recognize the need to obtain a large data base on the specific types and frequencies of mutations induced by a given substance in mammalian cells. We further recognize that these data should be obtained from different regions of a functioning mammalian gene within cell types that are representative of the specific cell types in vivo in which mutation and/or carcinogenesis occur. Thus, we have been developing an in vitro human cell assay which both detects mutagens and determine the frequency at which they cause specific transitions, transversions or frameshifts in transfected, site-specific mutated, mouse adenine phosphoribosyltransferase (APRT) genes. This gene was chosen because of its sensitive selectable characteristics. The specifically mutated mouse APRT genes can, in principle, be introduced into differentiated human cells which exhibit a spectrum of mutagen metabolizing enzymes similar to that observed in vivo. In addition, we have been developing immunological methods for directly detecting, in situ, mutagens that induce frameshifts.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA036897-04
Application #
3174517
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1984-06-01
Project End
1990-05-31
Budget Start
1987-06-10
Budget End
1988-05-31
Support Year
4
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Cincinnati
Department
Type
Schools of Medicine
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221
Zhu, Y; Bye, S; Stambrook, P J et al. (1995) Aflatoxin B1, 2-aminoanthracene, and 7,12-dimethylbenz[a]anthracene-induced frameshift mutations in human APRT. Environ Mol Mutagen 26:234-9
Zhu, Y; Bye, S; Stambrook, P J et al. (1994) Single-base deletion induced by benzo[a]pyrene diol epoxide at the adenine phosphoribosyltransferase locus in human fibrosarcoma cell lines. Mutat Res 321:73-9
Zhu, Y; Stambrook, P J; Tischfield, J A (1993) Loss of heterozygosity: the most frequent cause of recessive phenotype expression at the heterozygous human adenine phosphoribosyltransferase locus. Mol Carcinog 8:138-44
Chen, J; Sahota, A; Martin, G F et al. (1993) Analysis of germline and in vivo somatic mutations in the human adenine phosphoribosyltransferase gene: mutational hot spots at the intron 4 splice donor site and at codon 87. Mutat Res 287:217-25
Liu, H S; Scrable, H; Villaret, D B et al. (1992) Control of Ha-ras-mediated mammalian cell transformation by Escherichia coli regulatory elements. Cancer Res 52:983-9
Bertino, A M; Tischfield, J A; Stambrook, P J (1992) Reconstitution of an episomal mouse aprt gene as a consequence of recombination. Mol Gen Genet 232:24-32
Chen, J; Sahota, A; Laxdal, T et al. (1991) Identification of a single missense mutation in the adenine phosphoribosyltransferase (APRT) gene from five Icelandic patients and a British patient. Am J Hum Genet 49:1306-11
Sahota, A; Chen, J; Stambrook, P J et al. (1991) Mutational basis of adenine phosphoribosyltransferase deficiency. Adv Exp Med Biol 309B:73-6
Chen, J; Sahota, A; Stambrook, P J et al. (1991) Polymerase chain reaction amplification and sequence analysis of human mutant adenine phosphoribosyltransferase genes: the nature and frequency of errors caused by Taq DNA polymerase. Mutat Res 249:169-76
Sahota, A; Chen, J; Behzadian, M A et al. (1991) 2,8-Dihydroxyadenine lithiasis in a Japanese patient heterozygous at the adenine phosphoribosyltransferase locus. Am J Hum Genet 48:983-9

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