Matrix Degradation Assay to Measure the Ability of Tumor Cells to Degrade Extracellular Matrix

Rosa Fontana; Jing Yang

doi:10.1007/978-1-0716-1350-4_11

Matrix Degradation Assay to Measure the Ability of Tumor Cells to Degrade Extracellular Matrix

Methods Mol Biol. 2021:2294:151-163. doi: 10.1007/978-1-0716-1350-4_11.

Authors

Rosa Fontana¹, Jing Yang^{2

3}

Affiliations

¹ Department of Pharmacology, University of California, San Diego, La Jolla, CA, USA.
² Department of Pharmacology, University of California, San Diego, La Jolla, CA, USA. jingyang@ucsd.edu.
³ Moores Cancer Center, Department of Pediatrics, University of California, San Diego, La Jolla, CA, USA. jingyang@ucsd.edu.

PMID: 33742400
DOI: 10.1007/978-1-0716-1350-4_11

Abstract

During the metastatic process, carcinoma cells form invadopodia, F-actin enriched protrusive structures, to degrade the extracellular matrix (ECM) in order to invade the surrounding stroma and intravasate into the circulatory system. In this chapter, we describe the 2D-fluorescent matrix degradation assay, a highly sensitive and reproducible in vitro method used to measure invadopodia-mediated ECM degradation. We provide a detailed protocol on how to prepare the glass coverslips with fluorescent gelatin matrix and a standardized method to quantify gelatin degradation and invadopodia formation in order to evaluate cell invasion.

Keywords: Cancer metastasis; ECM; Fluorescent gelatin degradation assay; ImageJ; Invadopodia; Invadosome; Invasion; Podosome; Rosettes.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Cell Migration Assays / methods*
Cell Movement
Extracellular Matrix / metabolism*
Extracellular Matrix / pathology
Humans
Neoplasm Invasiveness / pathology*
Podosomes / metabolism
Podosomes / physiology

Abstract

Publication types

MeSH terms

Grants and funding