Summary
MDCK is a transformed cell line derived from normal dog kidney. It exhibits qualities of renal tubular epithelium and of papillary adenocarcinoma. On solid substrates, MDCK forms confluent monolayers that are studded with multicellular blisters and cordlike structures. Ouabain, at a concentration of 4×10−8 m, abolishes blister formation within 24 hr; withdrawal of the drug brings about reappearance of the blisters. Ouabain-sensitive ATPase activity has been demonstrated in these cells by biochemical measurement. These findings suggest that blister formation in MDCK monolayer growth is associated with active transport of solutes accompanied by water and that 4×10−8 m of ouabain interferes with these transport systems. As an additional early effect, the drug decreases the frequency of mitosis.
The exposure of MDCK monolayers to ouabain for several weeks results in profound cytological alterations. A marked eosinophilic stippling of the periphery of chronically exposed cells is seen with the light microscope. After prolonged exposure to ouabain, the cell population of MDCK monolayers becomes clearly heterogeneous, with frequent giant cells and foci of small, hyperchromatic proliferating cells. The small cells may indicate the appearance of ouabain-resistant strains.
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References
Shannon, J. E. (Ed.) 1972.Registry of Animal Cell Lines, The American Type Culture Collection. pp. CCL 34.
Leighton, J., Z. Brada, L. W. Estes, and G. Justh. 1969. Secretory activity and oncogenicity of a cell line (MDCK) derived from canine kidney. Science 163: 472–473.
Leighton, J., L. W. Estes, S. Mansukhani, and Z. Brada. 1970. A cell line derived from normal dog kidney (MDCK) exhibiting qualities of papillary adenocarcinoma and of renal tubular epithelium. Cancer 26: 1022–1028.
Leighton, J. 1969. Propagation of cancer: targets for future chemotherapy. Cancer Res. 29: 2457–2465.
Abaza, N. A., and J. Leighton. 1972. Effects of ouabain on a cell line (MDCK) derived from normal dog kidney (abstr.). Fed. Proc. 31: 626.
Schultz, S. G., and P. F. Curran. 1970. Coupled transport of sodium and organic solutes. Physiol. Rev. 50: 637–718.
Fiske, C. H., and Y. Subbarow. 1925. Calorimetric determination of phosphorus. J. Biol. Chem. 66: 375–400.
McGrath, C. M. 1971. Replication of mammary virus in tumor cell cultures: dependence on hormone-induced cellular organization. J. Natl. Cancer Inst. 47: 455–467.
McGrath, C. M., and P. M. Blair. 1970. Immunofluorescent localization of mamary tumor virus antigens in mammary tumor cells in culture. Cancer Res. 30: 1963–1968.
Kleinzeller, A. 1960. The role of potassium and calcium in the regulation of metabolism in kidney cortex slices. In: A. Kleinzeller, and A. Kotyk (Eds.),Membrane Transport and Metabolism. Academic Press, New York, pp. 527–542.
Lamb, J. F., and M. G. A. MacKinnon. 1967. Potassium influx in cultured cells in the presence of ouabain. J. Physiol. 191: 33P-34P.
Lamb, J. F., and M. G. A. MacKinnon. 1971. Effect of ouabain and metabolic inhibitors on the Na and K movements and nucleotide contents in L cells. J. Physiol. 213: 665–682.
Lamb, J. F., and D. McCall. 1970. The effects of prolonged treatment with ouabain on Na, K and Ca levels in cultured cells. J. Physiol. 206: 33P-34P.
McCall, D., J. F. Lamb, and T. D. V. Lawrie. 1972. Influence of extracellular potassium concentration on ouabain effect in human cultured cells. Br. Heart J. 34: 206.
Skou, J. C. 1957. The influence of some cations on an adenosine triphosphatase from peripheral nerves. Biochim. Biophys. Acta 23: 394–401.
Yoshida, H., T. Nukada, and H. Fujisawa. 1961. Effect of ouabain on ion transport and metabolic turnover of phospholipid of brain slices. Biochim. Biophys. Acta 48: 614–615.
Till, J. E., R. M. Baker, D. M. Brunette, V. Ling, L. H. Thompson, and J. A. Wright. 1973. Genetic regulation of membrane function in mammalian cells in culture. Fed. Proc. 32: 29–33.
Thompson, L. H., and R. M. Baker. 1973. Isolation of mutants of cultured mammalian cells. In: D. M. Prescott (Ed.),Methods in in Cell Biology, Vol. VI. Academic Press, New York, pp. 209–281.
Quastel, M. R., and J. G. Kaplan. 1968. Inhibition by ouabain of human lymphocyte transformation induced by phytohemagglutinin in vitro. Nature 219: 198–200.
Quastel, M. R., and J. G. Kaplan. 1970. Lymphocyte stimulation: The effect of ouabain on nucleic acid and protein synthesis. Exp. Cell Res. 62: 407–420.
Quastel, M. R., J. G. Kaplan, and J. D. Abbatt, 1971. Inhibition by ouabain of spontaneous DNA synthesis in acute leukaemic leucocytes in vitro. Lancet 1: 801–802.
Grahame-Smith, D. G., and A. G. Hibble. 1971. Inhibition of ouabain of spontaneous DNA synthesis in acute leukaemic leucocytes in vitro. Lancet 1: 1026.
Shiratori, O. 1967. Growth inhibitory effect of cardiac glycosides and aglycones on neoplastic cells: in vitro and in vivo studies. Gann 58: 521–528.
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This investigation was part of a dissertation submitted to the University of Pittsburgh in partial fulfillment of the requirements for the degree of Doctor of Philosophy.
This study was supported by NIH-NCI Research Grants CA 13219 and CA 14137, by Contract G-72-3858, and by NIH Fellowship FO 3 DE 426160 from the National Institutes of Health.
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Abaza, N.A., Leighton, J. & Schultz, S.G. Effects of ouabain on the function and structure of a cell line (MDCK) derived from canine kidney. In Vitro 10, 172–183 (1974). https://doi.org/10.1007/BF02615230
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DOI: https://doi.org/10.1007/BF02615230