Summary
Matrix invasion by a tumor cell requires the degradation of components in the extracellular matrix (ECM) as one of the initial steps in the metastatic process. Tumors cells achieve ECM invasion primarily through the overexpression of matrix metalloproteinases (MMPs), a family of enzymes that function to degrade ECM proteins. In this chapter, an in vitro collagen degradation assay and a modified collagen invasion assay system are described. The collagen degradation assay is a simple method to measure the ability of tumor cells to degrade type I collagen, the main constituent of the stromal compartment, in a 3-D matrix environment. The modified collagen invasion assay system enables researchers to study the effects of transient overexpression and/or targeted knockdown (as with siRNAs) of a given gene on collagen invasion of tumor cells in a real-time format.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Sternlicht MD, Werb Z. (2001) How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol 17, 463–516
Puente XS, Sanchez LM, Overall CM, Lopez-Otin C. (2003) Human and mouse proteases: a comparative genomic approach. Nat Rev Genet 4(7), 544–558
Egeblad M, Werb Z. (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2(3), 161–174
McCawley LJ, Matrisian LM. (2001) Matrix metalloproteinases: they're not just for matrix anymore! Curr Opin Cell Biol 13(5), 534–540
Folgueras AR, Pendas AM, Sanchez LM, Lopez-Otin C. (2004) Matrix metalloproteinases in cancer: from new functions to improved inhibition strategies. Int J Dev Biol 48(5–6), 411–424
Seiki M, Koshikawa N, Yana I. (2003) Role of pericellular proteolysis by membrane-type 1 matrix metalloproteinase in cancer invasion and angiogenesis. Cancer Metastasis Rev 22(2–3), 129–143
Overall CM, Lopez-Otin C. (2002) Strategies for MMP inhibition in cancer: innovations for the post-trial era. Nat Rev Cancer 2(9), 657–672
Butler TP, Gullino PM. (1975) Quantitation of cell shedding into efferent blood of mammary adenocarcinoma. Cancer Res 35(3), 512–516
Bogenrieder T, Herlyn M. (2003) Axis of evil: molecular mechanisms of cancer metastasis. Oncogene 22(42), 6524–6536
Chambers AF, Groom AC, MacDonald IC. (2002) Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer 2(8), 563–572
Fidler IJ. (2002) Critical determinants of metastasis. Semin Cancer Biol 12(2), 89–96
van der Rest M, Garrone R. (1991) Collagen family of proteins. Faseb J 5(13), 2814–2823
Gross J, Nagai Y. (1965) Specific degradation of the collagen molecule by tadpole col-lagenolytic enzyme. Proc Natl Acad Sci USA 54(4), 1197–1204
Lauer-Fields JL, Fields GB. (2002) Triple-helical peptide analysis of collagenolytic protease activity. Biol Chem 383(7–8), 1095–1105
Gross J, Kirk D. (1958) The heat precipitation of collagen from neutral salt solutions: some rate-regulating factors. J Biol Chem 233(2), 355–360
Gross J, Lapiere CM. (1962) Collageno-lytic activity in amphibian tissues: a tissue culture assay. Proc Natl Acad Sci USA 48, 1014–1022
Petrella BL, Brinckerhoff CE. (2006) Tumor cell invasion of von Hippel Lindau renal cell carcinoma cells is mediated by membrane type-1 matrix metalloproteinase. Mol Cancer 5, 66
Wyatt CA, Geoghegan JC, Brinckerhoff CE. (2005) Short hairpin RNA-mediated inhibition of matrix metalloproteinase-1 in MDA-231 cells: effects on matrix destruction and tumor growth. Cancer Res 65(23), 11101– 11108
Barrett AJ, Starkey PM. (1973) The interaction of alpha 2-macroglobulin with pro-teinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism. Biochem J 133(4), 709–724
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Petrella, B.L. (2009). Assessment of Local Proteolytic Milieu as a Factor in Tumor Invasiveness and Metastasis Formation: In Vitro Collagen Degradation and Invasion Assays. In: Kozlov, S.V. (eds) Inflammation and Cancer. Methods in Molecular Biology™, vol 511. Humana Press. https://doi.org/10.1007/978-1-59745-447-6_3
Download citation
DOI: https://doi.org/10.1007/978-1-59745-447-6_3
Publisher Name: Humana Press
Print ISBN: 978-1-934115-14-5
Online ISBN: 978-1-59745-447-6
eBook Packages: Springer Protocols