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Originally published In Press as doi:10.1074/jbc.M000237200 on March 20, 2000

J. Biol. Chem., Vol. 275, Issue 22, 17094-17099, June 2, 2000
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Stimulation of DNA Synthesis and Cell Proliferation of Human Mammary Myoepithelial-like Cells by Hepatocyte Growth Factor/Scatter Factor Depends on Heparan Sulfate Proteoglycans and Sustained Phosphorylation of Mitogen-activated Protein Kinases p42/44*

Nicolas SergeantDagger , Malcolm Lyon§, Philip S. RudlandDagger , David G. FernigDagger , and Maryse DeleheddeDagger

From the Dagger  School of Biological Sciences, University of Liverpool, Liverpool L69 7ZB and the § Cancer Research Campaign Department of Medical Oncology, University of Manchester, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, United Kingdom

Hepatocyte growth factor/scatter factor (HGF/SF) is a heparan/dermatan sulfate-binding growth factor produced by stromal cells that acts as a paracrine effector on neighboring epithelia. HGF/SF stimulated DNA synthesis in human mammary (Huma) 109 myoepithelial-like cells grown on collagen I and fibronectin substrata but not when grown on plastic. Dual phosphorylation of mitogen-activated protein kinases (p42/44MAPK) was required for this stimulation of DNA synthesis. In Huma 109 cells cultured on plastic, HGF/SF stimulated a transient phosphorylation of p42/44MAPK, which reached a maximum at 10 min after addition of the growth factor and returned to near basal levels after 20 min. In contrast, the phosphorylation of p42/44MAPK stimulated by HGF/SF in cells cultured on collagen I or fibronectin was sustained over 45 min. In Huma 109 cells deficient in sulfated glycosaminoglycans, HGF/SF failed to stimulate p42/44MAPK phosphorylation or DNA synthesis on any substratum, even when soluble heparan sulfate proteoglycans purified from the cells or from the culture medium were added. However, HGF/SF stimulated DNA synthesis and a sustained phosphorylation of p42/44MAPK in sulfated glycosaminoglycan-deficient Huma 109 cells plated on a substratum of medium HSPGs but not cell HSPGs. The HGF/SF-induced proliferation is thus highly dependent on heparan sulfate proteoglycans in myoepithelial-like cells.

* This work was supported by the Association for International Cancer Research, the Cancer Research Campaign, the Cancer and Polio Research Fund, and the North West Cancer Research Fund.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: School of Biological Sciences, Life Science Bldg., Crown St., University of Liverpool, Liverpool L69 7BZ, UK. Tel.: 44-151-794-4388; Fax: 44-151-794-4349; E-mail: dgfernig@liv.ac.uk.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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