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© 1986 Oxford University Press

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Cytotoxicity, cellular transformation and DNA adducts in normal human diploid fibroblasts exposed to 1-nitrosopyrene, a reduced derivative of the environmental contaminant, 1-nitropyrene

Frederick A. Beland , Martin Ribovich 1, Paul C. Howard 2 , Robert H. Heflich , Ponnamma Kurian 1 and George E. Milo 1

National Center for Toxicological Research Jefferson, AR 72079, USA
1Comprehensive Cancer Center, The Ohio State University Columbus, OH 43210, USA

2Present address: Department of Environmental Health Sciences, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA

Treatment of normal human diploid fibroblasts with 0.3 — 22 µM 1-nitrosopyrene resulted in a dose-dependent decrease in relative cloning efficiency and an increase in anchorage-independent growth in soft agar. When compared to previous experiments, 1-nitrosopyrene was 10- to 20-fold more cytotoxic and 5- to 10-fold more potent at inducing morphological transformation than 1-nitropyrene. Incubation of the fibroblasts with 8 µM 1-nitropyrene in the presence of xanthine oxidase, a mammalian nitroreductase, resulted in the formation of one major DNA adduct, N-(deoxyguanosin-8-yI)-1-aminopyrene, at a level of 1.1 adducts per 106 nucleo-tides. Fibroblasts treated with 1-nitrosopyrene formed the same DNA adduct; however, only a 0.3 µM concentration was required to give 0.7 adducts per 106 nucleotides in the fibro-blast DNA. These data indicate that a limiting step in the cellular toxicity and transformation of normal human diploid fibroblasts by 1-nitropyrene is the initial reduction to 1-nitrosopyrene.


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