Experiments were performed to determine the relative effects of glycosaminoglycans and extracellular matrix components alone or in association with various substrates, including extracellular matrix, on the proliferation of rat rhabdomyosarcoma (RMS) cell lines of different metastatic potential and nontumorigenic rat myoblast L6 cells. The assays used various substrates: tissue culture plastic, type I and IV collagen, fibronectin, laminin and extracellular matrix deposited by corneal endothelial cells. In control experiments, tumor cells grew faster on fibronectin and extracellular matrix than on the other substrates, and their proliferation rate was decreased slightly by laminin. Collagens were growth-inhibitory only for the highly metastatic line. The proliferation rate of L6 myoblasts was not greatly affected by the different substrates. The addition of exogenous glycosaminoglycans to the culture medium modified cell proliferation on the various substrates. Heparin inhibited the growth of all the cell lines tested, independent of the substrate. When cultured on laminin substrate the proliferation rates of the cell lines were depressed by addition of heparan sulfate to the medium, and this effect was more pronounced in the metastatic RMS lines. Chondroitin sulfate and dermatan sulfate enhanced the growth rates of the tumorigenic cells when cultured on collagen type I surfaces. Hydrocortisone, which induces myogenic differentiation, decreased the cell proliferation rates of all the cell lines tested and intensified the inhibitory effects of heparin when added simultaneously to the culture medium. The results showed that glycosaminoglycans and other matrix components can affect the proliferation rates of rhabdomyosarcoma cell lines.
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Becker, M., Moczar, E., Korach, S., Lascaux, V., and Poupon, M.-F., 1985, Relationship between in-vitro effects and in-vivo control of metastasis induced by hydrocortisone in a rat rhabdomyosarcoma model. Treatment of Metastasis: Problems and Prospects, edited by K. Hellmann and S. A. Eccles (London and Philadelphia: Taylor & Francis), pp. 183–186.
Becker, M., Poupon, M.-F., and Courtois, Y., 1984, Differential effect of EGF, FGF and EDGF on various metastatic cloned cell lines derived from a primary rat rhabdomyosarcoma. Lymphokine Research, 44, 2864–2868.
Bhavanandan, V. P., and Davidson, E. A., 1975, Mucopolysaccharides associated with nuclei of cultured mammalian cells. Proceedings of the National Academy of Sciences, U.S.A., 72, 2032–2036.
Coombe, D. R., Parish, C. R., Ramshaw, I. A., and Snowden, J. M., 1987, Analysis of the inhibition of tumour metastasis by sulphated polysaccharides. International Journal of Cancer, 39, 82–88.
Crum, R., Szabo, S., and Folkman, J., 1985, A new class of steroids inhibits angiogenesis in the presence of heparin or a heparin fragment. Science, 230, 1375–1378.
David, G., and Van Den Berghe, H., 1983, Transformed mouse mammary epithelial cells synthesize undersulfated basement membrane proteoglycan. Journal of Biological Chemistry, 258, 7338–7344.
Fedarko, N. S., and Conrad, H. E., 1986, A unique heparan sulfate in the nuclei of hepatocytes: structural changes with the growth state of the cells. Journal of Cell Biology, 102, 587–599.
Folkman, J., Langer, R., and Linhart, R. J., 1983, Angiogenesis inhibition and tumor regression caused by heparin fragment in the presence of cortisone. Science, 221, 719–725.
Glimelius, B., Norling, B., Westermark, B., and Wasteson, A., 1978, Composition and distribution of Glycosaminoglycans in culture of human normal and malignant filial cells. Biochemical Journal, 172, 443–456.
Gosporadowicz, D., Delgado, D., and Vlodavsky, I., 1980, Permissive effect of the extracellular matrix on cell proliferation in vitro. Proceedings of the National Academy of Sciences, U.S.A., 77, 4094–4098.
Grinnel, R., 1978, Cellular adhesiveness and biological characterization of active fragments of the adhesive glycoprotein fibronectin. International Review of Cytology, 53, 65–144.
Kleinman, H. K., Cannon, F. B., Laurie, G. W., Hassel, J. R., Aumailley, M., Terranova, V. P., Martin, G. R., and Dubois-Dalcq, M., 1985, Biological activities of laminin. Journal of Cellular Biochemistry, 27, 317–325.
Knox, P., and Wells, P., 1984, Cell adhesion and proteoglycans. Journal of Cell Science, 40, 77–88.
Knudson, W., Biswas, C., and Toole, B. P., 1984, Stimulation of glycosaminoglycan production in murine tumors. Journal of Cellular Biochemistry, 25, 183–196.
Laterra, J., Ansbacher, R., and Culp, L. A., 1980, Glycosaminoglycans that bind coldinsoluble globulin in cell-substratum sites of murine fibroblasts. Proceedings of the National Academy of Sciences, U.S.A., 77, 6662–6667.
Laterra, J., Silbert, S. E., and Culp, L. A., 1983, Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin. Journal of Cell Biology, 96, 112–117.
Leibovicz, M. I., and Leubutez, S. A., 1982, Effect of sodium butyrate on messenger RNA populations in myogenic cells in culture. Differentiation, 22, 106–112.
Maniglia, C. A., Gomez, J. J., Luikart, S. D., and Sartorelli, A. C., 1985, Glycosaminoglycan production and distribution in cloned B16 murine melanoma cell lines exhibiting different lung colony-forming efficiencies. Journal of the National Cancer Institute, 75, 111–120.
Matuoka, K., Mitsui, Y., and Muroka, S.-I., 1984, Heparan sulfate enhances growth of transformed human cells. Cell Structure and Functions, 9, 357–367.
Miketo, L. J., and Culp, L. A., 1983, Proteoglycans from the substratum adhesion sites of MSV-transformed BALB/C 3T3 cells. Archives of Biochemistry and Biophysics, 221, 447–455.
Milas, L., Hunter, N., and Basic, I., 1985, Treatment with cortisone, heparin or hexuronyl hexosaminoglycan of murine tumors and their lung deposits. Clinical and Experimental Metastasis, 3, 247–255.
Moczar, E., Tassin, J., and Courtois, Y., 1983, Interaction of bovine epithelial lens cells with extracellular matrix and eye-derived growth factor. III. Control of glycoprotein and proteoglycan synthesis. Experimental Cell Research, 149, 95–104.
Nakamura, T., Matsunaga, E., and Shinhai, H., 1983, Isolation and some structural analyses of a proteodermatan sulphate from calf skin. Biochemical Journal, 213, 289–296.
Pauli, B. U., Schwartz, D. E., Thonar, E. J., and Kuettner, K. E., 1983, Tumor invasion and host extracellular matrix. Cancer Metastasis Review, 2, 129–152.
Piepkorn, M., Carney, H., and Linker, A., 1985, Growth-related variations in the glycosaminoglycan synthesis of ultraviolet light-induced murine cutaneous fibrosarcoma cells. Laboratory Investigations, 53, 187–193.
Reddan, J. R., Dziedzic, D. C., McGee, S. J., Dehart, D. J., and Sackman, J. E., 1982, Influence of extracellular matrix, laminin and fibronectin on the growth of cultured mammalian lens epithelial cells. Extracellular Matrix, edited by S. Hawkes and J. L. Wang (New York: Academic Press), pp. 171–176.
Redini, F., Moczar, E., and Poupon, M.-F., 1986, Cell surface glycosaminoglycans of rat rhabdomyosarcoma lines with different metastatic potentials and of non-malignant rat myoblasts. Biochimica et Biophysica Acta, 883, 98–105.
Rollins, B. J., and Culp, L. A., 1979, Preliminary characterization of the proteoglycan in the substrate adhesion sites of normal and virus-transformed murine cells. Biochemistry, 18, 5621–5628.
Bong, G. G., Alessandri, G., and Sindelar, W. F., 1986, Inhibition of tumor angiogenesis by hexuronyl hexosaminoglycan sulfate. Cancer, 57, 586–590.
Ruoslahti, E., Engvall, E., and Hayman, E. G., 1981, Fibronectin: current concepts of its structure and functions. Collagen Related Research, 1, 95–128.
Sweeney, F. L., Pot-Deprun, J., Poupon, M.-F., and Chouroulinkov, I., 1982, Heterogeneity of the growth and metastatic behavior of cloned cell lines derived from a primary rhabdomyosarcoma. Cancer Research, 42, 3776–3800.
Tassin, J., Jacquemin, E., and Courtois, Y., 1983, Interaction of bovine epithelial lens cells with extracellular matrix and eye derived growth factor. I. Effects on short term adhesion and on long term organization of the culture. Experimental Cell Research, 149, 69–84.
Timpl, R., Rhode, H., Ristelli, L., Ott, Y., Robey, P. J., and Martin, G. R., 1982, Laminin. Methods of Enzymology, 82, 831–838.
Turley, E. A., 1984, Proteoglycans and cell adhesion. Cancer and Metastasis Review, 3, 325–339.
Vlodavsky, I., Folkman, J., Sullivan, R., Fridman, R., Ishai-Michaeli, R., Sasse, J., and Klagsbrun, M., 1987, Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. Proceedings of the National Academy of Sciences, U.S.A., 84, 2292–2296.
Weiss, R. E., and Reddi, A. H., 1981, Appearance of fibronectin during the differentiation of cartilage, bone and bone marrow. Journal of Cell Biology, 88, 630–636.
Wright, T. C., Johnstone, T. V., Castellot, J. J., and Karnovsky, M. J., 1985, Inhibition of rat cervical epithelial cell growth by heparin and its reversal by EGF. Journal of Cell Physiology, 125, 499–506.
Yaffe, D., 1968, Retention of differentiation potentialities during prolonged cultivation of myogenic cells. Proceedings of the National Academy of Sciences, U.S.A., 61, 477–481.
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Redini, F., Moczar, E. & Poupon, M.F. Effects of glycosaminoglycans and extracellular matrix components on metastatic rat rhabdomyosarcoma tumor and myoblast cell proliferation. Clin Exp Metast 8, 491–502 (1990). https://doi.org/10.1007/BF00058158
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DOI: https://doi.org/10.1007/BF00058158