To test the hypothesis that activation of bladder carcinogens occurs intracellularly by enzymes associated with the transitional epithelial cells of the bladder, the metabolic capabilities of these cells will be investigated. The bioactivation of the human urinary bladder carcinogen, 4-aminobiophenyl(ABP); 4-nitrobiphenyl(NBP); and the experimental bladder carcinogen N-[4-(5-nitro-2-furyl)-2-thiazolyl] formamide (FANFT) will be examined using normal human and rat urothelial cells. The nature of intermediate(s) that are activated by the urothelial cells will be assessed by testing the cytosolic and microsomal fractions for certain enzymatic activities such as N-oxidation, nitroreduction, N-glucuronidation, sulfation, acetylation, deacetylation and N--O transacylation; and by identifying the DNA-carcinogen adducts. The DNA-carcinogen adducts will be isolated from cultured human urothelial cells after treatment with tritium labeled ABP, NBP, 4-nitrosobiphenyl, N-hydroxylaminobiphenyl or its N-glucuronide conjugate. The profiles of the adducts generated from cultured cells will be compared with those obtained by chemical interaction of N-hydroxylaminobiphenyl or its N-glucuronide derivative with calf thymus DNA in vitro at pH 5.0. The formation of, and biological reactivities of the proximate metabolites will be assessed by urothelium-mediated mutagenicity tests in Salmonella typhimurium strains. A long range objective of this project is to understand which of the adducts and enzymes play a determinate role in the initiation of human cancers caused by ABP and its related analogs; and to find out the metabolic factors responsible for the observed differences in susceptibility to ABP-induced bladder carcinogenesis between rats and humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES003509-03
Application #
3250851
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1984-08-01
Project End
1987-07-31
Budget Start
1986-08-01
Budget End
1987-07-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Schools of Medicine
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Hatcher, J F; Yamamoto, K; Ichikawa, M et al. (1995) Metabolic reduction of novel 3,4-dichloro-5-nitrofurans in Salmonella typhimurium. Environ Mol Mutagen 25:58-66
Hatcher, J F; Swaminathan, S (1995) 32P-postlabeling analysis of adducts generated by peroxidase-mediated binding of N-hydroxy-4-acetylaminobiphenyl to DNA. Carcinogenesis 16:2149-57
Hatcher, J F; Swaminathan, S (1995) Detection of deoxyadenosine-4-aminobiphenyl adduct in DNA of human uroepithelial cells treated with N-hydroxy-4-aminobiphenyl following nuclease P1 enrichment and 32P-postlabeling analysis. Carcinogenesis 16:295-301
Swaminathan, S; Frederickson, S M; Hatcher, J F (1994) Metabolic activation of N-hydroxy-4-acetylaminobiphenyl by cultured human breast epithelial cell line MCF 10A. Carcinogenesis 15:611-7
Kloth, M T; Gee, R L; Messing, E M et al. (1994) Expression of N-acetyltransferase (NAT) in cultured human uroepithelial cells. Carcinogenesis 15:2781-7
Frederickson, S M; Messing, E M; Reznikoff, C A et al. (1994) Relationship between in vivo acetylator phenotypes and cytosolic N-acetyltransferase and O-acetyltransferase activities in human uroepithelial cells. Cancer Epidemiol Biomarkers Prev 3:25-32
Hatcher, J F; Rao, K P; Swaminathan, S (1993) Mutagenic activation of 4-aminobiphenyl and its N-hydroxy derivatives by microsomes from cultured human uroepithelial cells. Mutagenesis 8:113-20
Swaminathan, S; Hatcher, J F (1992) Comparison of acyltransferase-mediated mutagenicity and nucleic acid binding of N-acetoxy-4-acetylaminobiphenyl by hepatic and bladder microsomes from rats and dogs. Environ Mol Mutagen 20:61-72
Frederickson, S M; Hatcher, J F; Reznikoff, C A et al. (1992) Acetyl transferase-mediated metabolic activation of N-hydroxy-4-aminobiphenyl by human uroepithelial cells. Carcinogenesis 13:955-61
Swaminathan, S; Reznikoff, C A (1992) Metabolism and nucleic acid binding of N-hydroxy-4-acetylaminobiphenyl and N-acetoxy-4-acetylaminobiphenyl by cultured human uroepithelial cells. Cancer Res 52:3286-94

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