title: Single cell tracking assay reveals an opposite effect of selective small non-peptidic α5β1 or αvβ3/β5 integrin antagonists in U87MG glioma cells.
abstract: BACKGROUND
  Integrins are extracellular matrix receptors involved in several pathologies. Despite homologies between the RGD-binding α5β1 and αvβ3 integrins, selective small antagonists for each heterodimer have been proposed. Herein, we evaluated the effects of such small antagonists in a cellular context, the U87MG cell line, which express both integrins. The aim of the study was to determine if fibronectin-binding integrin antagonists are able to impact on cell adhesion and migration in relationships with their defined affinity and selectivity for α5β1 and αvβ3/β5 purified integrins.
  
  
  METHODS
  Small antagonists were either selective for α5β1 integrin, for αvβ3/β5 integrin or non-selective. U87MG cell adhesion was evaluated on fibronectin or vitronectin. Migration assays included wound healing recovery and single cell tracking experiments. U87MG cells stably manipulated for the expression of α5 integrin subunit were used to explore the impact of α5β1 integrin in the biological assays.
  
  
  RESULTS
  U87MG cell adhesion on fibronectin or vitronectin was respectively dependent on α5β1 or αvβ3/β5 integrin. Wound healing migration was dependent on both integrins. However U87MG single cell migration was highly dependent on α5β1 integrin and was inhibited selectively by α5β1 integrin antagonists but increased by αvβ3/β5 integrin antagonists.
  
  
  CONCLUSIONS
  We provide a rationale for testing new integrin ligands in a cell-based assay to characterize more directly their potential inhibitory effects on integrin cellular functions.
  
  
  CONCLUSIONS
  Our data highlight a single cell tracking assay as a powerful cell-based test which may help to characterize true functional integrin antagonists that block α5β1 integrin-dependent cell migration.
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France; CNRS UMR 7200, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Ray
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Schaffner
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Hana
authors.initials: H
authors.lastname: Janouskova
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Noulet
authors.affiliation: CNRS UMR 7200, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Rognan
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Lelong-Rebel
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Choulier
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Anne-Florence
authors.initials: AF
authors.lastname: Blandin
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Lehmann
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Martin
authors.affiliation: Institute for Advanced Study, Technische Universität München, Department Chemie, Lichtenbergerstrasse 4, Garching, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Kapp
authors.affiliation: Institute for Advanced Study, Technische Universität München, Department Chemie, Lichtenbergerstrasse 4, Garching, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Neubauer
authors.affiliation: Institute for Advanced Study, Technische Universität München, Department Chemie, Lichtenbergerstrasse 4, Garching, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Rechenmacher
authors.affiliation: Institute for Advanced Study, Technische Universität München, Department Chemie, Lichtenbergerstrasse 4, Garching, Germany; Chemistry Department, Faculty of Science, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia.
authors.forename: Horst
authors.initials: H
authors.lastname: Kessler
authors.affiliation: CNRS UMR 7213, Université de Strasbourg, Faculté de Pharmacie, 74 route du Rhin, 67401 Illkirch, France. Electronic address: monique.dontenwill@unistra.fr.
authors.forename: Monique
authors.initials: M
authors.lastname: Dontenwill
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D004354:Drug Screening Assays, Antitumor; D005910:Glioma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D039081:Integrin alpha5beta1 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D039302:Integrin alphaVbeta3 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D039641:Integrin beta Chains; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D039081:Integrin alpha5beta1; D039302:Integrin alphaVbeta3; D039641:Integrin beta Chains; D009363:Neoplasm Proteins; C084344:integrin beta5
keywords: Cell adhesion; Cell migration; Small antagonist; α5β1 integrin; αvβ3/β5 integrin