A Fluorescent Gelatin Degradation Assay to Study Melanoma Breakdown of Extracellular Matrix

Methods Mol Biol. 2021:2265:47-63. doi: 10.1007/978-1-0716-1205-7_3.

Abstract

In order to protrude within a dense tissue, tumor cells have to develop the ability to digest the extracellular matrix (ECM). Melanoma cells, similarly to other types of tumor cells, form invadopodia, membranous invaginations rich in filamentous actin and several other proteins including matrix metalloproteinases (MMPs). MMPs degrade ECM structural proteins such as collagens, fibronectin, or laminin. Here we describe an assay that allows the detection of gelatinase activity exhibited by tumor cells under 2D conditions and methods to present obtained data in both a quantitative and a qualitative manner.

Keywords: Extracellular matrix (ECM) degradation; Gelatin-fluorescein degradation assay; Invadopodia; Invasion; Matrix metalloproteinases (MMPs); Melanoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Cell Culture Techniques / methods
  • Cell Line, Tumor
  • Extracellular Matrix / enzymology*
  • Extracellular Matrix / metabolism
  • Extracellular Matrix / pathology
  • Fluorescence
  • Gelatin / metabolism*
  • Gelatinases / metabolism
  • Humans
  • Matrix Metalloproteinases / metabolism
  • Melanoma / enzymology*
  • Melanoma / pathology
  • Microscopy, Confocal / methods*
  • Optical Imaging
  • Podosomes / enzymology
  • Podosomes / metabolism
  • Podosomes / pathology

Substances

  • Actins
  • Gelatin
  • Gelatinases
  • Matrix Metalloproteinases