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  • Title: Induction of ouabain-resistant mutants by chemical carcinogens in rat prostate epithelial cells.
    Author: Link KH, Heidelberger C, Landolph JR.
    Journal: Environ Mutagen; 1983; 5(1):33-48. PubMed ID: 6299722.
    Abstract:
    We determined optimal conditions to quantitatively select ouabain-resistant (Ouar) mutants induced by chemical carcinogens in a rat prostate epithelial cell line (RPYK). These conditions included selection of Ouar mutants in 3 mM ouabain, an expression time of two days following a two-day treatment with carcinogens, and a reseeding density of 2 X 10(5) mutagenized cells per 100 mm dish to select mutants in ouabain. Ouar mutants induced by N-methyl-N'-nitro-N'-nitrosoguanidine (MNNG) remained stably Ouar when passaged in nonselective medium. Hemicyst formation, a characteristic of epithelial cells, was reversibly inhibited by ouabain in wild-type cells and was resistant to ouabain in Ouar cells. The direct-acting carcinogens MNNG and methylazoxymethanol-acetate (MAMA) and the environmentally widespread procarcinogens aflatoxin B1 and benzo(a)pyrene increased the frequency of Ouar mutants in RPYK cells. The procedures developed here now make it possible to detect some environmental carcinogens likely to cause prostate cancer by virtue of their ability to mutate cultured prostate epithelial RPYK cells. The sensitivity of the RPYK cell line to aflatoxin-induced cytotoxicity and mutagenesis also makes it a useful cell system in which to study enzymes governing the conversion of aflatoxin to genotoxic metabolites.
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