Journal of Cell Science, Vol 97, Issue 2 373-383, Copyright © 1990 by Company of Biologists

JOURNAL ARTICLES

The pattern of metalloproteinase expression by corneal fibroblasts is altered by passage in cell culture

ME Fini and MT Girard
Eye Research Institute, Harvard Medical School, Boston, Massachusetts 02114.

We have examined the pattern of expression of four different matrix metalloproteinases (MMPs), collagenase, stromelysin, 92 kD gelatinase, and 72 kD gelatinase, by primary and passaged cultures of rabbit corneal fibroblasts. Primary cultures of this cell type have previously been shown to reproduce the normal tissue regulation of collagenase expression. We demonstrate qualitative and quantitative changes in the pattern of MMP expression as the cells are passaged in culture. Only a single MMP, 72 kD gelatinase, is constitutively expressed by primary fibroblast cultures. Phorbol myristate acetate (PMA) treatment upregulates expression of 72 kD gelatinase and turns on the expression of collagenase and stromelysin, as well as 92 kD gelatinase. However, the degree to which MMP expression is induced is minimal. Cells subcultured but a single time constitutively produce not only 72 kD gelatinase, but also collagenase and stromelysin. In addition, PMA treatment upregulates expression of collagenase, stromelysin and 92 kD gelatinase to high levels. In contrast, the expression of 72 kD gelatinase is repressed by treatment of passaged cell cultures with PMA. Our data indicate that the cell does not simply turn the MMP genes on or off, as a group, in response to various agents, but that it has the capacity for fine control over which MMPs are expressed and the degree to which each is expressed. Changes in MMP protein expression induced by PMA treatment are correlated with changes in specific mRNA levels in passaged cultures. The kinetics of mRNA accumulation suggest that the MMP genes can respond to multiple intracellular signals initiated in a temporal cascade by PMA. It is the combined effects of the individual signals on the accumulation of specific mRNAs that must determine the ultimate pattern of MMP protein expression. The distinct patterns of MMP expression produced by primary and passaged cell cultures may be analogous to patterns of expression that might occur under particular in vivo conditions.

This article has been cited by other articles:

				Y. Lu, K. Fukuda, Y. Liu, N. Kumagai, and T. Nishida Dexamethasone Inhibition of IL-1-Induced Collagen Degradation by Corneal Fibroblasts in Three-Dimensional Culture Invest. Ophthalmol. Vis. Sci., September 1, 2004; 45(9): 2998 - 3004. [Abstract] [Full Text] [PDF]

				Y. Lu, K. Fukuda, K. Seki, Y. Nakamura, N. Kumagai, and T. Nishida Inhibition by Triptolide of IL-1-Induced Collagen Degradation by Corneal Fibroblasts Invest. Ophthalmol. Vis. Sci., December 1, 2003; 44(12): 5082 - 5088. [Abstract] [Full Text] [PDF]

				Q. Li, K. Fukuda, Y. Lu, Y. Nakamura, T.-i. Chikama, N. Kumagai, and T. Nishida Enhancement by neutrophils of collagen degradation by corneal fibroblasts J. Leukoc. Biol., September 1, 2003; 74(3): 412 - 419. [Abstract] [Full Text] [PDF]

				C. Ngamkitidechakul, J. M. Burke, W. J. O'Brien, and S. S. Twining Maspin: Synthesis by Human Cornea and Regulation of In Vitro Stromal Cell Adhesion to Extracellular Matrix Invest. Ophthalmol. Vis. Sci., December 1, 2001; 42(13): 3135 - 3141. [Abstract] [Full Text] [PDF]

				O. Maltseva, P. Folger, D. Zekaria, S. Petridou, and S. K. Masur Fibroblast Growth Factor Reversal of the Corneal Myofibroblast Phenotype Invest. Ophthalmol. Vis. Sci., October 1, 2001; 42(11): 2490 - 2495. [Abstract] [Full Text] [PDF]

				T. Nagano, J.-L. Hao, M. Nakamura, N. Kumagai, M. Abe, T. Nakazawa, and T. Nishida Stimulatory Effect of Pseudomonal Elastase on Collagen Degradation by Cultured Keratocytes Invest. Ophthalmol. Vis. Sci., May 1, 2001; 42(6): 1247 - 1253. [Abstract] [Full Text]

				D.-Q. Li, S.-B. Lee, Z. Gunja-Smith, Y. Liu, A. Solomon, D. Meller, and S. C. G. Tseng Overexpression of Collagenase (MMP-1) and Stromelysin (MMP-3) by Pterygium Head Fibroblasts Arch Ophthalmol, January 1, 2001; 119(1): 71 - 80. [Abstract] [Full Text] [PDF]

				C. J. Long, M. R. Roth, E. S. Tasheva, M. Funderburgh, R. Smit, G. W. Conrad, and J. L. Funderburgh Fibroblast Growth Factor-2 Promotes Keratan Sulfate Proteoglycan Expression by Keratocytes in Vitro J. Biol. Chem., April 28, 2000; 275(18): 13918 - 13923. [Abstract] [Full Text] [PDF]

				R. Mohan, J. Sivak, P. Ashton, L. A. Russo, B. Q. Pham, N. Kasahara, M. B. Raizman, and M. E. Fini Curcuminoids Inhibit the Angiogenic Response Stimulated by Fibroblast Growth Factor-2, Including Expression of Matrix Metalloproteinase Gelatinase B J. Biol. Chem., March 31, 2000; 275(14): 10405 - 10412. [Abstract] [Full Text] [PDF]

				K. S. Spirin, A. V. Ljubimov, R. Castellon, O. Wiedoeft, M. Marano, D. Sheppard, M. C. Kenney, and D. J. Brown Analysis of Gene Expression in Human Bullous Keratopathy Corneas Containing Limiting Amounts of RNA Invest. Ophthalmol. Vis. Sci., December 1, 1999; 40(13): 3108 - 3115. [Abstract] [Full Text] [PDF]

				J. R. Cook, M. K. Mody, and M. E. Fini Failure to Activate Transcription Factor NF-{kappa}B in Corneal Stromal Cells (Keratocytes) Invest. Ophthalmol. Vis. Sci., December 1, 1999; 40(13): 3122 - 3131. [Abstract] [Full Text] [PDF]

				H. Schnaper, J. Kopp, A. Poncelet, S. Hubchak, W. Stetler-Stevenson, P. Klotman, and H. Kleinman Increased expression of extracellular matrix proteins and decreased expression of matrix proteases after serial passage of glomerular mesangial cells J. Cell Sci., January 10, 1996; 109(10): 2521 - 2528. [Abstract] [PDF]

				B. Himelstein, R Canete-Soler, E. Bernhard, and R. Muschel Induction of fibroblast 92 kDa gelatinase/type IV collagenase expression by direct contact with metastatic tumor cells J. Cell Sci., January 2, 1994; 107(2): 477 - 486. [Abstract] [PDF]