Abstract
Analysing cell migration and invasion is of interest to many investigators as they mimic a part of physiological or pathological events. In this chapter, methods to analyse MMP-dependent 2D and 3D cell migration are described in detail. To study 2D cell migration, the phagokinetic track motility assay can be used. To study 3D cell migration, Matrigel invasion assay or microcarrier beads invasion assay can be used to obtain quantitative results.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Egeblad, M. and Werb, Z. (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2, 161–174.
Itoh, Y. (2006) MT1-MMP: a key regulator of cell migration in tissue. IUBMB Life 58, 589–596.
Wolf, K., Wu, Y. I., Liu, Y., Geiger, J., Tam, E., Overall, C., Stack, M. S., and Friedl, P. (2007) Multi-step pericellular proteolysis controls the transition from individual to collective cancer cell invasion. Nat Cell Biol 9, 893–904.
Sato, H., Takino, T., Okada, Y., Cao, J., Shinagawa, A., Yamamoto, E., and Seiki, M. (1994) A matrix metalloproteinase expressed on the surface of invasive tumour cells. Nature 370, 61–65.
Itoh, Y. and Seiki, M. (2006) MT1-MMP: a potent modifier of pericellular microenvironment. J Cell Physiol 206, 1–8.
Albrecht-Buehler, G. (1977) The phagokinetic tracks of 3T3 cells. Cell 11, 395–404.
Kajita, M., Itoh, Y., Chiba, T., Mori, H., Okada, A., Kinoh, H., and Seiki, M. (2001) Membrane-type 1 matrix metalloproteinase cleaves CD44 and promotes cell migration. J Cell Biol 153, 893–904.
Dietrich, F. and Lelkes, P. I. (2006) Fine-tuning of a three-dimensional microcarrier-based angiogenesis assay for the analysis of endothelial-mesenchymal cell co-cultures in fibrin and collagen gels. Angiogenesis 9, 111–125.
Naor, D., Sionov, R. V., and Ish-Shalom, D. (1997) CD44: structure, function, and association with the malignant process. Adv Cancer Res 71, 241–319.
Weber, G. F., Bronson, R. T., Ilagan, J., Cantor, H., Schmits, R., and Mak, T. W. (2002) Absence of the CD44 gene prevents sarcoma metastasis. Cancer Res 62, 2281–2286.
Nagano, O., Murakami, D., Hartmann, D., De Strooper, B., Saftig, P., Iwatsubo, T., Nakajima, M., Shinohara, M., and Saya, H. (2004) Cell-matrix interaction via CD44 is independently regulated by different metalloproteinases activated in response to extracellular Ca(2+) influx and PKC activation. J Cell Biol 165, 893–902.
Anilkumar, N., Uekita, T., Couchman, J. R., Nagase, H., Seiki, M., and Itoh, Y. (2005) Palmitoylation at Cys574 is essential for MT1-MMP to promote cell migration. FASEB J 19, 1326–1328.
Uekita, T., Itoh, Y., Yana, I., Ohno, H., and Seiki, M. (2001) Cytoplasmic tail-dependent internalization of membrane-type 1 matrix metalloproteinase is important for its invasion-promoting activity. J Cell Biol 155, 1345–1356.
Bafetti, L. M., Young, T. N., Itoh, Y., and Stack, M. S. (1998) Intact vitronectin induces matrix metalloproteinase-2 and tissue inhibitor of metalloproteinases-2 expression and enhanced cellular invasion by melanoma cells. J Biol Chem 273, 143–149.
Itoh, Y., Takamura, A., Ito, N., Maru, Y., Sato, H., Suenaga, N., Aoki, T., and Seiki, M. (2001) Homophilic complex formation of MT1-MMP facilitates proMMP-2 activation on the cell surface and promotes tumor cell invasion. Embo J 20, 4782–4793.
Melchiori, A., Albini, A., Ray, J. M., and Stetler-Stevenson, W. G. (1992) Inhibition of tumor cell invasion by a highly conserved peptide sequence from the matrix metalloproteinase enzyme prosegment. Cancer Res 52, 2353–2356.
Sato, K., Ebihara, T., Adachi, E., Kawashima, S., Hattori, S., and Irie, S. (2000) Possible involvement of aminotelopeptide in self-assembly and thermal stability of collagen I as revealed by its removal with proteases. J Biol Chem 275, 25870–25875.
Acknowledgments
This work is supported by Cancer research UK project grant C1507/A2670. We thank Rob Visse for critical reading of the manuscript.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Palmisano, R., Itoh, Y. (2010). Analysis of MMP-Dependent Cell Migration and Invasion. In: Clark, I. (eds) Matrix Metalloproteinase Protocols. Methods in Molecular Biology, vol 622. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-299-5_23
Download citation
DOI: https://doi.org/10.1007/978-1-60327-299-5_23
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-298-8
Online ISBN: 978-1-60327-299-5
eBook Packages: Springer Protocols