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Endocrinology Vol. 141, No. 10 3764-3773
Copyright © 2000 by The Endocrine Society

ARTICLES

Functional Significance of MMP-9 in Tumor Necrosis Factor-Induced Proliferation and Branching Morphogenesis of Mammary Epithelial Cells1

Ping-Ping H. Lee, Jiuan-Jiuan Hwang, Gillian Murphy and Margot M. Ip

Department of Pharmacology and Therapeutics, Grace Center Drug Center, Roswell Park Cancer Institute (P.-P.H.L., M.M.I.), Buffalo, New York 14263; Institute of Physiology, National Yang-Ming University (J.J.H.), Taipei, Taiwan; and School of Biological Sciences, University of East Anglia (G.M.), Norwich, Norfolk, United Kingdom

Address all correspondence and requests for reprints to: Dr. Margot M. Ip, Grace Cancer Drug Center, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, New York 14263. E-mail: margot.ip{at}RoswellPark.org

Tissue remodeling is a key process involved in normal mammary gland development, with matrix metalloproteinases (MMPs) playing an important role in this process. Our laboratory has demonstrated that tumor necrosis factor (TNF) stimulates branching morphogenesis of mammary epithelial cells (MEC) within a reconstituted basement membrane. Studies were therefore undertaken to determine whether MMPs might mediate the effects of TNF. Using a primary culture model in which rat MEC grow three-dimensionally within a reconstituted basement membrane, we found that TNF stimulated secretion of MMP-9 but not MMP-2. To determine whether MMP-9 was involved in TNF-induced proliferation and branching morphogenesis, we used a peptide containing the prodomain sequence of MMPs and two MMP inhibitors. Both the prodomain peptide (5 x 10-4–10-3 M), as well as BB-94 (10-8–10-5 M) and CGS 27023A (10-6–10-5 M), inhibited TNF-induced proliferation and branching morphogenesis in a concentration-dependent manner. Finally, to verify the specific requirement for MMP-9, we demonstrated that an MMP-9 neutralizing antibody blocked TNF-induced proliferation and branching morphogenesis. Together, these data suggest that TNF-regulated MMP-9 may play a role in the controlled invasion of the fad pad that occurs during normal mammary gland development and that misregulation of MMP-9 may contribute to the invasiveness of breast cancer.




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