title: Boyden Chamber Assay to Study of Cell Migration Induced by Metalloprotease Cleaved-CD95L.
abstract: CD95 receptor, also called Fas or Apo-1, is a member of the tumor necrosis factor receptors (TNF-R) superfamily (Itoh and Nagata, J Biol Chem 268:10932-10937, 1993). Its cognate ligand, CD95L, is a transmembrane cytokine, which can be cleaved by metalloproteases (Matsuno et al., J Rheumatol 28:22-28, 2001; Vargo-Gogola et al., Arch Biochem Biophys 408:155-161, 2002; Kiaei et al., Exp Neurol 205:74-81, 2007; Schulte et al., Cell Death Differ 14:1040-1049, 2007) releasing a soluble ligand into the bloodstream. Recent work has shown that this metalloprotease-cleaved CD95L (cl-CD95L) is involved in carcinogenesis (Malleter et al., Cancer Res 73:6711-6721, 2013). Cl-CD95L also fuels the inflammatory process in patients affected by systemic lupus erythematosus by promoting the accumulation of activated T lymphocytes in enflamed organs (Tauzin et al., PLoS Biol 9:e1001090, 2011). This chapter aims at describing the methodology used to measure the chemoattractive effect of cl-CD95L on human cancer cells and lymphocytes.
authors.affiliation: Centre Eugène Marquis, rue Bataille Flandres Dunkerque, 35042, Rennes, France.|INSERM ERL440-OSS, Equipe Labellisée Ligue Contre Le Cancer, Rennes, France.|Université de Rennes-1, 2 av. du Prof. Léon Bernard, 35043, Rennes, France.
authors.forename: Amanda
authors.initials: A
authors.lastname: Poissonnier
authors.affiliation: Centre Eugène Marquis, rue Bataille Flandres Dunkerque, 35042, Rennes, France. patrick.legembre@inserm.fr.|INSERM ERL440-OSS, Equipe Labellisée Ligue Contre Le Cancer, Rennes, France. patrick.legembre@inserm.fr.|Université de Rennes-1, 2 av. du Prof. Léon Bernard, 35043, Rennes, France. patrick.legembre@inserm.fr.
authors.forename: Patrick
authors.initials: P
authors.lastname: Legembre
mesh_terms: D017209:Apoptosis; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D002465:Cell Movement; D042783:Endothelial Cells; D053222:Fas Ligand Protein / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes; D045726:Metalloproteases / Q000378:metabolism*; D011485:Protein Binding; D059748:Proteolysis; D057705:Transendothelial and Transepithelial Migration; D019014:fas Receptor / Q000378:metabolism
chemical_list: D053222:Fas Ligand Protein; D019014:fas Receptor; D045726:Metalloproteases
keywords: Boyden chamber; Cell migration; Cell motility; Fas; FasL; Metastasis; PI3K