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  • Title: Correlation between the loss of the transformed phenotype and an increase in superoxide dismutase activity in a revertant subclone of sarcoma virus-infected mammalian cells.
    Author: Fernandez-Pol JA, Hamilton PD, Klos DJ.
    Journal: Cancer Res; 1982 Feb; 42(2):609-17. PubMed ID: 6275983.
    Abstract:
    We have studied the effects of paraquat (methyl viologen), a herbicide that increases intracellular production of superoxide radical, on the viability of virus-transformed and nontransformed normal rat kidney (NRK) cells in culture. We have shown that a low concentration of paraquat (12.5 microM) is cytotoxic toward virus-transformed cell lines, including Kirsten sarcoma virus- and SV40-transformed NRK cells. The corresponding untransformed NRK cells were resistant to the same and a 4-fold higher concentration of paraquat. There was a good correlation between the susceptibility of transformed and untransformed cells to paraquat cytotoxicity and their ability to increase the superoxide dismutase (SOD) enzymatic activity. We found that paraquat is cytotoxic toward Kirsten sarcoma virus-transformed and SV40-transformed NRK cells which showed low intracellular SOD activity. The relationship between SOD activity and paraquat cytoxicity was strengthened by the finding that the tolerance of NRK cells to the drug was associated with high intracellular SOD activity. This report also describes the isolation of a revertant (revertant RE8G3) cell line derived from Kirsten sarcoma virus-transformed NRK cells after paraquat treatment which contains SOD activity at levels much higher than those found in NRK cells. This revertant is undistinguishable from NRK cells with respect to its lack of transformed cell properties. Not only are these cells normal morphologically but also they do not grow in soft agar, an in vitro property that closely correlates with in vivo tumorigenicity. Several biological and biochemical properties of RE8G3 cells, including growth characteristics, surface receptors for both transferrin and epidermal growth factor (EGF), and the EGF-dependent 32P phosphorylation of specific membrane polypeptides have been studied. The most interesting conclusion that can be drawn from these studies is that there is a correlation between loss of the transformed phenotype and an increase in both EGF receptors and EGF-dependent 32P phosphorylation of a m.w. 170,000 membrane-associated protein.
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