Phorbol ester binding was examined in two lines of murine fibrosarcoma cells. The two cell lines were isolated from the same parent tumor but respond differentially to stimulation with phorbol esters. In one of the lines, these agents stimulate a rapid attachment and spreading response and induce directional migration. The other cell line does not migrate in response to stimulation with phorbol esters and the attachment and spreading response is slow. The cell line which responds actively to phorbol ester stimulation is highly malignant when injected into syngeneic animals while the other line is of low tumorigenicity and is virtually non-metastatic. In spite of these differences, both lines were found in the present study to bind [3H]4β-phorbol-12β, 13α-dibutyrate in a receptor-mediated fashion. The characteristics of binding were virtually identical between the two cell lines. In additional studies, arachidonic acid metabolism was examined in the same two lines. In the highly responsive line, PMA stimulated a rapid release of [3H]arachidonic acid and its conversion into cyclooxygenase and lipoxygenase products. In the less-responsive line, PMA stimulated a slower release of [3H]arachidonic acid from prelabeled cells. The quantity of arachidonic acid metabolites produced was also much less. These studies suggest that the disparity between the two cell lines in their response to phorbol ester stimulation is not the result of differences in the initial interaction between the cells and ligand but may result from alterations in their signal transductance mechanism. This may be the result of inherent differences in capacity for arachidonic acid metabolism.
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Aswanikumar, S., Corcoran, B., Schiffmann, E., Day, A. R., Freer, R. J., Showell, H. J., Becker, E. L., and Pert, C. B., 1977, Demonstration of a receptor on rabbit neutrophils for chemotactic peptides. Biochemical and Biophysical Research Communications, 74, 8111–817.
Bishop, R., Martinez, R., Weber, M. J., Blackshear, P. J., Beatty, S., Lim, R., and Herschman, H. R., 1985, Protein phosphorylation in a tetradecanoyl phorbol acetate-nonproliferative variant of 3T3 cells. Molecular and Cellular Biology, 5, 2231–2237.
Blumberg, P. M., Butler-Gralla, E., and Herschman, H. R., 1981, Analysis of phorbol ester receptors in phorbol ester unresponsiveness 3T3 cell variants. Biochemical and Biophysical Research Communications, 102, 818–823.
Butler-Gralla, E., Taplitz, S., and Herschman, H. R., 1983, 12-O-tetradecanoyl phorbol-13-acetate stimulates release of arachidonic acid, prostaglandin E2 and prostaglandin F2α from TPA nonproliferative variants of 3T3 cells. Biochemical and Biophysical Research Communications, 111, 194–199.
Chang, J., 1983, Characterization of a high-affinity receptor for phorbol esters in rat alveolar macrophages. Inflammation, 7, 15–23.
Clark, P. R. H., and Varani, J., 1984, Phorbol ester binding to chemotactically responding and nonresponding Walker 256 carcinosarcoma cells. Cancer Research, 44, 4967–4971.
Driedger, P. E., and Blumberg, P. M., 1980, Specific binding of phorbol ester tumor promoters. Proceedings of the National Academy of Sciences, U.S.A., 77, 567–571.
Fligiel, S. E. G., Perone, P., and Varani, J., 1985, Arachidonic acid metabolism in murine fibrosarcoma cells with differing in vivo and in vitro characteristics. International Journal of Cancer, 36, 383–388.
Folch, J., Lees, M., Sloane Stanley, G. N., 1957, A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497–509.
Goldman, D. W., and Goetzl, E. J., 1983, Mediation and modulation of immediate hypersensitivity and inflammation by products of the oxygenation of arachidonic acid. Handbook of Inflammation, Vol. 4, edited by P. A. Ward (Amsterdam: Elsevier), pp. 163–188.
Horowitz, A. D., Greenbaum, E., and Weinstein, I. B., 1981, Identification of receptors for phorbol ester tumor promoters in intact mammalian cells and an inhibitor of receptor binding in biological fluids. Proceedings of the National Academy of Sciences, U.S.A., 78, 2315–2319.
Jimenez, A., Clingan, L. D., and Rudland, P. S., 1975, Initiation of cell proliferation in cultured mouse fibroblasts by prostaglandin F2α. Proceedings of the National Academy of Sciences, U.S.A., 72, 2724–2725.
Lam, W. C., Delikatny, E. J., Orr, F. W., Wass, J., Varani, J., and Ward, P. A., 1981, The chemotactic response of tumor cells: A model for cancer metastasis. American Journal of Pathology, 104, 69–76.
Marasco, W. A. Ward, P. A., Feltner, D. E., and Varani, J., 1985, Chemotactic factor binding by metastatic tumor cells: Evidence for a formylpeptide receptor on a nonmyelogenous cell. Journal of Cell Science, 73, 121–134.
Munson, P. J., and Rodgard, D., 1980, LIGAND: A versatile computerized approach for characterization of ligand-binding systems. Analytical Biochemistry, 107, 220–239.
Ozaki, T., Yoshida, K., Ushijima, K., Hayashi, H., 1971, Studies on the mechanisms of invasion in cancer. II. In vivo effects of a factor chemotactic for cancer cells. International Journal of Cancer, 7, 93–100.
Rayner, D. C., Orr, F. W., and Shiu, R. P., 1985, Binding of formyl peptides to Walker 256 carcinosarcoma cells and the chemotactic response of these cells. Cancer Research, 45, 2288–2293.
Sando, J. J., Hilfiker, M. L., Piacentini, M. J., and Laufer, T. M., 1982, Identification of phorbol ester receptors in T-cell growth factor-producing and nonproducing EL4 mouse thymoma cells. Cancer Research, 42, 1676–1682.
Sando, J. J., Hilfiker, M. L., Salomon, D. S., and Farror, J. J., 1981, Specific receptors for phorbol esters in lymphoid cell populations: Role in enhanced production of T-cell growth factor. Proceedings of the National Academy of Sciences, U.S.A., 78, 1189–1193.
Scatchard, G., 1949, The attraction of proteins for small molecules and ions. Annals of the New York Academy of Sciences, 51, 660–672.
Shoyab, M., and Todaro, G. J., 1980, Specific, high-affinity cell membrane receptors for biologically-active phorbol and ingenol esters. Nature, 288, 451–455.
Skipski, V. P., Peterson, R. F., and Barclay, M., 1964, Quantitative analysis of phospholipids by thin layer chromatography. Biochemical Journal, 90, 374–378.
Varani, J., 1982, Chemotaxis of metastatic tumor cells. Cancer Metastasis Reviews, 1, 17–28.
Varani, J., Wass, J., Piontek, G., and Ward, P. A., 1981, Chemotactic factor-induced adherence of tumor cells. Cell Biology International Reports, 5, 525–530.
Varani, J., and Fantone, J. C., 1982, Phorbol myristate acetate-induced adherence of Walker 256 carcinosarcoma cells. Cancer Research, 42, 190–197.
Varani, J., Fligiel, S. E. G., and Perone, P., 1985, Directional motility in strongly malignant murine tumor cells. International Journal of Cancer, 35, 559–564.
Varani, J., Lovett, E. J., Wicha, M., Malinoff, H., and McCoy, J. P., 1983, Cell surface α-d-galactopyranosyl end groups: use as a marker in the isolation of murine tumor cell lines with varying malignant potential. Journal of the National Cancer Institute, 71, 1281–1286.
Williams, L. T., Snyderman, R., Pike, M. C., and Lefkowitz, R. J., 1977, Specific receptor sites for chemotactic peptides on human polymorphonuclear leukocytes. Proceedings of the National Academy of Sciences, U.S.A., 74, 1204–1208.
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Batchev, A.C., Riser, B.L., Hellner, E.G. et al. Phorbol ester binding and phorol ester-induced arachidonic acid metabolism in a highly responsive murine fibrosarcoma cell line and in a less-responsive variant. Clin Exp Metast 4, 51–61 (1986). https://doi.org/10.1007/BF00053473
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DOI: https://doi.org/10.1007/BF00053473