Summary
We previously demonstrated that human keratinocyte cultures proliferate in the absence of polypeptide growth factors (autonomous growth) and that this autonomous growth is blocked by interaction of heparin with a human keratinocyte-derived autocrine factor (KAF) which we identified as amphiregulin (AR). In the present study, we demonstrate that sulfated polysaccharides other than heparin (low and high molecular weight dextran sulfates) also inhibit the AR-mediated autonomous proliferation of human keratinocytes. Furthermore, sulfated polysaccharides such as high and low molecular weight dextran sulfates, heparan sulfate and, to a lesser extent, chondroitin sulfates B and C were also shown to be inhibitors of human keratinocyte-derived AR (k-d AR)-stimulated DNA synthesis in quiescent murine AKR-2B cell cultures. Our results demonstrate that sulfation of polysaccharides is required for AR inhibitory activity, and that several sulfated polysaccharides (other than heparin) can act as inhibitors of AR-mediated autonomous proliferation in human epidermal keratinocytes and as inhibitors of k-d AR-mediated mitogenic activity in AKR-2B cells.
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References
Ciardiello, F.; Kim, N.; Saeki, T., et al. Differential expression of epidermal growth factor-related proteins in human colorectal tumors. Proc. Natl. Acad. Sci. USA 88:7792–7796; 1991.
Ciardiello, F.; Donno, R.; Kim, N., et al. Expression of cripto, a novel gene of the epidermal growth factor gene family, leads to in vitro transformation of a normal mouse mammary epithelial cell line. Cancer Res. 51:1051–1054; 1991.
Ciccodicola, A.; Donno, R.; Obici, S., et al. Molecular characterization of a gene of the ‘EGF family’ expressed in undifferentiated human NTERA2 teratocarcinoma cells. EMBO J. 8:1987–1991; 1989.
Clowes, A. W.; Karnovsky, M. J. Suppression by heparin of smooth muscle proliferation in injured arteries. Nature 265:625–626; 1977.
Cook, P. W.; Mattox, P. A.; Keeble, W. W., et al. A heparin sulfateregulated human keratinocyte autocrine factor is similar or identical to amphiregulin. Mol. Cell. Biol. 11:2547–2557; 1991.
Cook, P. W.; Pittelkow, M. R.; Shipley, G. D. Growth factor-independent proliferation of normal human neonatal keratinocytes and production of autocrine- and paracrine-acting mitogenic factors. J. Cell. Physiol. 146:277–289; 1991.
Higashiyama, S.; Abraham, J. A.; Miller, J., et al. A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science 251:936–939; 1991.
Hook, M.; Kjellen, L.; Johansson, S. Cell surface glycosaminoglycan. Ann. Rev. Biochem. 53:847–869; 1984.
Imamura, T.; Mitsui, Y. Heparan sulfate and heparin as a potentiator or a suppressor of growth of normal and transformed vascular endothelial cells. Exp. Cell Res. 172:92–100; 1987.
Majesky, M. W.; Schwartz, S. M.; Clowes, M. M., et al. Heparin regulates smooth muscle S phase entry in the injured rat carotid artery. Circ. Res. 61:296–300; 1987.
Plowman, G. D.; Green, J. M.; McDonald, V. L., et al. The amphiregulin gene encodes a novel epidermal growth factor-related protein with tumor-inhibitory activity. Mol. Cell. Biol. 10:1969–1981; 1990.
Robertson, P. L.; Goldstein, G. W. Heparin inhibits the growth of astrocytes in vitro. Brain Res. 447:341–345; 1988.
Rosen, E. M.; Goldberg, I. D.; Kacinski, B. M., et al. Smooth muscle releases an epithelial cell scatter factor which binds to heparin. In Vitro Cell. Dev. Biol. 25:163–173; 1989.
Shipley, G. D.; Childs, C. B.; Volkenant, M. E., et al. Differential effects of epidermal growth factor, transforming growth factor, and insulin on DNA and protein synthesis and morphology in serum-free cultures of AKR-2B cells. Cancer Res. 44:710–716; 1984.
Shipley, G. D.; Ham, R. G. Improved medium and culture conditions for clonal growth with minimal serum protein and for enhanced serum-free survival of swiss 3T3 cells. In Vitro Cell. Dev. Biol. 17:656–670; 1981.
Shipley, G. D.; Keeble, W. W.; Hendrickson, J. E., et al. Growth of normal human keratinocytes and fibroblasts in serum-free medium is stimulated by acidic and basic fibroblast growth factor. J. Cell. Physiol. 138:511–518; 1989.
Shipley, G. D.; Pittelkow, M. R. Control of growth and differentiation in vitro of human prokeratinocytes cultured in serum-free medium. Arch. Dermatol. 123:1541a-1544a; 1987.
Shoyab, M.; McDonald, V. M.; Bradley, J. G., et al. Amphiregulin: a bifunctional growth-modulating glycoprotein produced by the phorbol 12-myristate 13-acetate-treated human breast adenocarcinoma cell line MCF-7. Proc. Natl. Acad. Sci. USA 85:6528–6532; 1988.
Shoyab, M.; Plowman, G. D.; McDonald, V. M., et al. Structure and function of human amphiregulin: a member of the epidermal growth factor family. Science 243:1074–1076; 1989.
Wille, J. J. Jr.; Pittelkow, M. R.; Shipley, G. D., et al. Integrated control of growth and differentiation of normal human prokeratinocytes cultured in serum free medium clonal analysis, growth kinetics, and cell cycle studies. J. Cell. Physiol. 121:31–44; 1984.
Wright, T. C. Jr.; Castellot, J. J. Jr.; Petitou, M., et al. Structural determinants of heparings growth inhibitory activity. J. Biol. Chem. 264:1534–1542; 1989.
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Cook, P.W., Mattox, P.A., Keeble, W.W. et al. Inhibition of autonomous human keratinocyte proliferation and amphiregulin mitogenic activity by sulfated polysaccharides. In Vitro Cell Dev Biol - Animal 28, 218–222 (1992). https://doi.org/10.1007/BF02631096
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DOI: https://doi.org/10.1007/BF02631096