Surprisingly little is known about the activities responsible for polycyclic aromatic hydrocarbon (PAH) metabolism in the epidermis. Our recent studies suggest that proliferating and differentiating keratinocytes have different capacities for PAH metabolism. The overall goal of this proposal is to determine whether epidermal PAH metabolism and tumorigenesis are influenced by the differentiation status of the tissue. Specifically, we will a) characterize the capacities of basal, proliferating and differentiating keratinocytes for PAH metabolism, b) characterize the enzymatic activities responsible for metabolism, and c) relate these activities to PAH-dependent epidermal tumorigenesis. Analyses will be made in primary cultures of proliferating and differentiating murine keratinocytes, and freshly isolated suspensions of keratinocytes separated by density gradient centrifugation into basal and differentiating cell populations. Capacities for oxidation of the PAHs, DMBA, BP and BP-7,8-diol will be measured by monitoring their metabolism or comparing mutant frequencies obtained in a keratinocyte- mediated mutation assay. Metabolite profiles will provide an indication of whether there are quantitative or qualitative differences in metabolizing activities, and studies with (+)BP-7,8- diol will provide estimates of mixed-function monooxygenase (MFO) and non-MFO (peroxidase-like) mediated metabolism. Antibodies to specific cytochrome P-450 species will be used in immunoblot studies to quantify these components of the MFO system. In vivo skin tumor studies in conjunction with PAH-DNA adduct analyses, using (+)BP-7,8-diol as the initiator, will be used to assess the in vivo relevance of MFO and non-MFO activities in the various keratinocyte cell types to epidermal carcinogenesis. Lastly, the differentiation status of murine epidermis will be manipulated to determine its influence on in vivo PAH metabolism (measured as PAH-DNA adducts) and carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA040823-07
Application #
3181121
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1985-09-01
Project End
1992-02-29
Budget Start
1991-09-01
Budget End
1992-02-29
Support Year
7
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of Texas MD Anderson Cancer Center
Department
Type
Organized Research Units
DUNS #
001910777
City
Houston
State
TX
Country
United States
Zip Code
77030
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Reiners Jr, J J; Scholler, A; Bischer, P et al. (1993) Suppression of cytochrome P450 Cyp1a-1 induction in murine hepatoma 1c1c7 cells by 12-O-tetradecanoylphorbol-13-acetate and inhibitors of protein kinase C. Arch Biochem Biophys 301:449-54
Reiners Jr, J J; Cantu, A R; Scholler, A (1992) Phorbol ester-mediated suppression of cytochrome P450 Cyp1a-1 induction in murine skin: involvement of protein kinase C. Biochem Biophys Res Commun 186:970-6
Reiners Jr, J J; Cantu, A R; Thai, G et al. (1992) Differential expression of basal and hydrocarbon-induced cytochrome P-450 monooxygenase and quinone reductase activities in subpopulations of murine epidermal cells differing in their stages of differentiation. Drug Metab Dispos 20:360-6
Reiners Jr, J J; Pavone, A; Cantu, A R et al. (1992) Differential expression of cytochrome P-450 in proliferating and quiescent cultures of murine lung epithelial cells. Biochem Biophys Res Commun 183:193-8
Reiners Jr, J J; Cantu, A R; Pavone, A (1991) Distribution of constitutive and polycyclic aromatic hydrocarbon-induced cytochrome P-450 activities in murine epidermal cells that differ in their stages of differentiation. Prog Clin Biol Res 369:123-35
Reiners Jr, J J; Kodari, E; Cappel, R E et al. (1991) Assessment of the antioxidant/prooxidant status of murine skin following topical treatment with 12-O-tetradecanoylphorbol-13-acetate and throughout the ontogeny of skin cancer. Part II: Quantitation of glutathione and glutathione disulfide. Carcinogenesis 12:2345-52
Reiners Jr, J J; Thai, G; Rupp, T et al. (1991) Assessment of the antioxidant/prooxidant status of murine skin following topical treatment with 12-O-tetradecanoylphorbol-13-acetate and throughout the ontogeny of skin cancer. Part I: Quantitation of superoxide dismutase, catalase, glutathione peroxidase Carcinogenesis 12:2337-43
Reiners Jr, J J; Rupp, T; Conti, C J (1991) Modulation of xanthine dehydrogenase and oxidase activities during the hormonal induction of vaginal epithelial differentiation in ovariectomized mice. Differentiation 47:69-75

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