A Combined Phagocytosis and Fluorescent Substrate Degradation Assay to Simultaneously Assess Cell Migration and Substrate Degradation

Methods Mol Biol. 2016:1406:195-202. doi: 10.1007/978-1-4939-3444-7_17.

Abstract

In order to more rapidly define the mechanism by which certain drugs and compounds can influence cancer cell invasion, we have combined a traditional phagokinetic gold migration assay with a fluorescent substrate. The purpose of this dual assay is to provide a platform by which to simultaneously monitor proteolytic activity and cancer cell migratory ability, both of which are required for the crucial step of cancer cell invasion during metastasis. This assay allows for delineation of potential mechanisms of action a compound of interest has, as one can determine whether or not a cancer cell that is being treated with the potential drug has changes in proteolytic activity and/or migratory ability at the same time.

Keywords: Dual assay; Fluorescent substrate; Migration; Phagokinetic; Substrate degradation.

MeSH terms

  • Cell Line
  • Cell Migration Assays / methods*
  • Fluorescent Dyes / metabolism*
  • Gelatin / chemistry
  • Gold / chemistry
  • Phagocytosis*
  • Proteolysis
  • Spectrometry, Fluorescence
  • Time Factors
  • Xanthenes / chemistry

Substances

  • Fluorescent Dyes
  • Xanthenes
  • Gold
  • Texas red
  • Gelatin