title: Silencing of Eps8 inhibits in vitro angiogenesis.
abstract: OBJECTIVE
  Eps8 is an actin-binding protein which has been proposed as a regulator of cancer cell motility and invasion. However, nothing much is known about its contribution to the invasive properties of endothelial cells (ECs), and more generally to angiogenesis.
  
  
  METHODS
  Expression and silencing of Eps8 were evaluated by western blot analysis. The effect of Eps8 silencing on cell number and VEGF-induced signaling was tested with standard methods. Migration was evaluated by scratch wound assay and morphogenesis with 2-dimensional (2-D) tube formation and 3-dimensional (3-D) sprouting assays. Actin cytoskeleton was visualized by immunofluorescence.
  
  
  RESULTS
  We found that silencing of Eps8 profoundly affected the ability of human ECs to migrate and to undergo tube formation and sprouting in 2-D and 3-D in vitro assays, respectively. Notably, capillary-like outgrowth was strictly depending on Eps8 expression also in human tumor-derived ECs.
  
  
  CONCLUSIONS
  Our data demonstrate for the first time the involvement of Eps8 in the morphological processes required for in vitro angiogenesis, and suggest that this protein might represent a common target for the design of new anticancer drugs, acting at the same time on both tumor and endothelial cells.
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, Via Vanvitelli 32, 20129 Milano, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Cappellini
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, Via Vanvitelli 32, 20129 Milano, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Vanetti
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, Via Vanvitelli 32, 20129 Milano, Italy.
authors.forename: Lucia M
authors.initials: LM
authors.lastname: Vicentini
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, Università degli Studi di Milano, Via Vanvitelli 32, 20129 Milano, Italy. Electronic address: mgrazia.cattaneo@unimi.it.
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Cattaneo
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D015153:Blotting, Western; D002465:Cell Movement; D005455:Fluorescent Antibody Technique; D020868:Gene Silencing; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism*; D006801:Humans; D009389:Neovascularization, Pathologic / Q000235:genetics* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C489989:EPS8 protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Endothelial cells; Eps8; In vitro angiogenesis; Tumor-derived endothelial cells