Summary
Proteolytic degradation of extracellular matrix (ECM) is a critical step during cell invasion and tissue transmigration that is required for many physiological and pathological processes. Cellular structures that mediate cell adhesion to, degradation of, and invasion into ECM are invadopodia of transformed and tumor cells and podosomes of normal monocytic, endothelial, and smooth muscle cells. Detecting the ability of the cell to form invadopodia and podosomes and to degrade ECM is required for studying the invasive capability of the cell. We have developed ~50 nm thick fluorescent gelatin matrices that provide a rapid, sensitive, and reliable in vitro system for detection of invadopodia and podosomes, and measurements of the extent of ECM degradation. In this chapter, we provide a detailed protocol for preparation of thin fluorescent gelatin matrices and for evaluation of the results from this degradation assay.
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References
Linder, S. (2007) The matrix corroded: podosomes and invadopodia in extracellular matrix degradation. Trends. Cell. Biol. 17(3), 107–117
Coopman, P. J., Do, M. T., Thompson, E. W., and Mueller, S. C. (1998) Phagocytosis of cross-linked gelatin matrix by human breast carcinoma cells correlates with their invasive capacity. Clin. Cancer. Res. 4(2), 507–515
Thompson, E. W. Paik, S. Brunner, N. Sommers, C. L. Zugmaier, G. Clarke, R. Shima, T. B. Torri, J. Donahue, S. Lippman, M. E., et al. (1992) Association of increased basement membrane invasiveness with absence of estrogen receptor and expression of vimentin in human breast cancer cell lines. J. Cell Physiol. 150(3), 534–544
Chen, W. T. Olden, K. Bernard, B. A. Chu, F. F. (1984) Expression of transformation-associated protease(s) that degrade fibronectin at cell contact sites. J. Cell Biol. 98(4), 1546–1555
Chen, W. T. (1989) Proteolytic activity of specialized surface protrusions formed at rosette contact sites of transformed cells. J. Exp. Zool. 251(2), 167–185
Artym, V. V., Zhang, Y., Seillier-Moiseiwitsch, F., Yamada, K. M., Mueller, S. C. (2006) Dynamic interactions of cortactin and membrane type 1 matrix metalloproteinase at invadopodia: defining the stages of invadopodia formation and function. Cancer. Res. 66(6), 3034–3043
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Artym, V., Yamada, K., Mueller, S. (2009). ECM Degradation Assays for Analyzing Local Cell Invasion. In: Even-Ram, S., Artym, V. (eds) Extracellular Matrix Protocols. Methods in Molecular Biology, vol 522. Humana Press. https://doi.org/10.1007/978-1-59745-413-1_15
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DOI: https://doi.org/10.1007/978-1-59745-413-1_15
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Publisher Name: Humana Press
Print ISBN: 978-1-58829-984-0
Online ISBN: 978-1-59745-413-1
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