title: Elastin peptides regulate HT-1080 fibrosarcoma cell migration and invasion through an Hsp90-dependent mechanism.
abstract: BACKGROUND
  The elastin-derived peptides (EDPs) exert protumoural activities by potentiating the secretion of matrix metalloproteinases (MMP) and the plasminogen-plasmin activating system. In the present paper, we studied heat-shock protein 90 (Hsp90) involvement in this mechanism.
  
  
  METHODS
  HT-1080 fibrosarcoma cell migration and invasion were studied in artificial wound assay and modified Boyden chamber assay, respectively. Heat-shock protein 90 was studied by western blot and immunofluorescence. Matrix metalloproteinase-2 and urokinase plasminogen activator (uPA) were studied by gelatin ± plasminogen zymography and immunofluorescence. Heat-shock protein 90 partners were studied by immunoprecipitation. Messenger RNA expression was studied using real-time PCR. Small interfering RNAs were used to confirm the essential role of Hsp90.
  
  
  RESULTS
  We showed that kappa-elastin and VGVAPG elastin hexapeptide stimulated Hsp90, pro-MMP-2 and uPA secretion within 6 h, whereas AGVPGLGVG and GRKRK peptides had no effect. No increase of mRNA level was observed. Heat-shock protein 90-specific inhibitors inhibit EDP-stimulated HT-1080 cell-invasive capacity and restrained EDP-stimulated pro-MMP-2 and uPA secretions. The inhibitory effect was reproduced by using Hsp90-blocking antibody or Hsp90 knockdown by siRNA. Heat-shock protein 90 interacted with and stabilised uPA and pro-MMP-2 in conditioned culture media of HT-1080 fibrosarcoma cells.
  
  
  CONCLUSIONS
  Taken together, our results demonstrate that EDPs exert protumoural activities through an Hsp90-dependent mechanism involving pro-MMP-2 and uPA.
authors.affiliation: 1] Laboratoire de Biochimie Médicale et Biologie Moléculaire, UMR CNRS/URCA 7369 MEDyC, UFR Médecine, Université de Reims Champagne Ardenne, 51 Rue Cognacq Jay, 51095 Reims, France [2] CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire - MEDyC, 51 Rue Cognacq Jay, 51095 Reims, France.
authors.forename: M
authors.initials: M
authors.lastname: Donet
authors.affiliation: 1] Laboratoire de Biochimie Médicale et Biologie Moléculaire, UMR CNRS/URCA 7369 MEDyC, UFR Médecine, Université de Reims Champagne Ardenne, 51 Rue Cognacq Jay, 51095 Reims, France [2] CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire - MEDyC, 51 Rue Cognacq Jay, 51095 Reims, France.
authors.forename: S
authors.initials: S
authors.lastname: Brassart-Pasco
authors.affiliation: 1] Laboratoire de Biochimie Médicale et Biologie Moléculaire, UMR CNRS/URCA 7369 MEDyC, UFR Médecine, Université de Reims Champagne Ardenne, 51 Rue Cognacq Jay, 51095 Reims, France [2] CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire - MEDyC, 51 Rue Cognacq Jay, 51095 Reims, France.
authors.forename: S
authors.initials: S
authors.lastname: Salesse
authors.affiliation: 1] Laboratoire de Biochimie Médicale et Biologie Moléculaire, UMR CNRS/URCA 7369 MEDyC, UFR Médecine, Université de Reims Champagne Ardenne, 51 Rue Cognacq Jay, 51095 Reims, France [2] CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire - MEDyC, 51 Rue Cognacq Jay, 51095 Reims, France [3] CHU de Reims, Laboratoire Central de Biochimie, 51092 Reims, France.
authors.forename: F-X
authors.initials: FX
authors.lastname: Maquart
authors.affiliation: 1] Laboratoire de Biochimie Médicale et Biologie Moléculaire, UMR CNRS/URCA 7369 MEDyC, UFR Médecine, Université de Reims Champagne Ardenne, 51 Rue Cognacq Jay, 51095 Reims, France [2] CNRS UMR 7369, Matrice Extracellulaire et Dynamique Cellulaire - MEDyC, 51 Rue Cognacq Jay, 51095 Reims, France.
authors.forename: B
authors.initials: B
authors.lastname: Brassart
mesh_terms: D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D004549:Elastin / Q000494:pharmacology*; D005354:Fibrosarcoma / Q000378:metabolism* / Q000473:pathology*; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D018841:HSP90 Heat-Shock Proteins; D010446:Peptide Fragments; D004549:Elastin; D014568:Urokinase-Type Plasminogen Activator; C471083:Col-1 peptide, human; D020778:Matrix Metalloproteinase 2