In Vitro Methods to Study the Modulation of Migration and Invasion by Sphingosine-1-Phosphate

Methods Mol Biol. 2018;1697:117-131. doi: 10.1007/7651_2017_51.

Abstract

Sphingosine-1-phosphate (S1P) is a bioactive lipid that modulates migratory behavior of cells during embryonic development. In addition, S1P might promote tumor progression by enhancing migratory ability and invasiveness of tumor cells. Migration is a complex process that implies cytoskeletal reorganization and formation of structures that enable cell movement. Besides having similar requirements than migration, invasion also involves proteolytic degradation of extracellular matrix (ECM). Matrix metalloproteases (MMPs) have been identified to break down components of the ECM, allowing cancer cells to spread out of the primary tumor. In this chapter, we will describe different techniques to study migration and invasion induced by S1P. To this end, we include detailed protocols of end-point assays to study migration/invasion, and zymography assay to analyze MMP-2 and MMP-9 activity that were standardized in our laboratory in human melanoma cell lines.

Keywords: Boyden Chamber; Invasion; Melanoma; Migration; Sphingosine-1-phosphate; Transwell; Wound healing; Zymography.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • In Vitro Techniques
  • Lysophospholipids / pharmacology*
  • Matrix Metalloproteinases / metabolism*
  • Melanoma / metabolism*
  • Neoplasm Invasiveness
  • Sphingosine / analogs & derivatives*
  • Sphingosine / pharmacology

Substances

  • Lysophospholipids
  • sphingosine 1-phosphate
  • Matrix Metalloproteinases
  • Sphingosine