Functional role of matrix metalloproteinases (MMPs) in mammary epithelial cell development.
The extracellular matrix (ECM) is an important regulator of mammary epithelial cell (MEC) function and is remodeled by matrix metalloproteinases (MMPs). To investigate the significance and regulation of MMP activity in normal MEC, we utilized a primary culture model in which rat MEC were grown three dimensionally within a reconstituted basement membrane (RBM) in defined serum-free medium. Zymograms of culture medium demonstrated that five major gelatinases of 97, 80, 74, 69, and 65 kDa were secreted by MEC and were distinct from gelatinases of RBM origin. Based on molecular weight, p-aminophenylmercuric acid activation, immunoblotting with MMP-specific antibodies, inhibition by EDTA, a peptide containing the prodomain sequence of MMP (TMRKPRCGNPDVAN) and two synthetic MMP inhibitors (BB-94 and CGS 27023A), these were classified as inactive and active forms of MMP-9 and MMP-2. The maximal MMP activities occurred when MEC were in a rapid proliferation and branching phase and declined after they underwent functional differentiation. Known regulators of MEC growth and differentiation were evaluated for their ability to modulate gelatinase activity in primary culture. Secretion of one or both MMPs was inhibited by EGF, TGFalpha, prolactin, and hydrocortisone and stimulated by progesterone. Furthermore, the functional significance of MMPs was demonstrated since three MMP inhibitors blocked branching morphogenesis elicited by the absence of hydrocortisone. Additionally, two synthetic MMP inhibitors not only inhibited epithelial cell growth but also inhibited normal alveolar development of the MEC. Finally, these drugs were found to enhance MMP secretion from MEC, although the activity of the secreted MMPs was inhibited as long as the drug was present.
Lee
P P
PP
Grace Center Drug Center, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263, USA.
Hwang
J J
JJ
Mead
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Ip
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MM
D000818:Animals; D048429:Cell Size / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D004815:Epidermal Growth Factor / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D018093:Gelatinases / Q000378:metabolism*; D006801:Humans; D006877:Hydroxamic Acids; D015151:Immunoblotting; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D051379:Mice; D009940:Organoids / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D010663:Phenylmercury Compounds / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D011719:Pyrazines; D051381:Rats; D017207:Rats, Sprague-Dawley; D013449:Sulfonamides; D013876:Thiophenes / Q000494:pharmacology; D016211:Transforming Growth Factor alpha / Q000378:metabolism
C097658:CGS 27023A; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D006877:Hydroxamic Acids; D061965:Matrix Metalloproteinase Inhibitors; D010455:Peptides; D010663:Phenylmercury Compounds; D011480:Protease Inhibitors; D011719:Pyrazines; D013449:Sulfonamides; D013876:Thiophenes; D016211:Transforming Growth Factor alpha; D010649:Phenylalanine; D004815:Epidermal Growth Factor; C080985:batimastat; D018093:Gelatinases; D020782:Matrix Metalloproteinases