title: Alpha-enolase (ENO1) controls alpha v/beta 3 integrin expression and regulates pancreatic cancer adhesion, invasion, and metastasis.
abstract: We have previously shown that in pancreatic ductal adenocarcinoma (PDA) cells, the glycolytic enzyme alpha-enolase (ENO1) also acts as a plasminogen receptor and promotes invasion and metastasis formation. Moreover, ENO1 silencing in PDA cells induces oxidative stress, senescence and profoundly modifies PDA cell metabolism. Although anti-ENO1 antibody inhibits PDA cell migration and invasion, little is known about the role of ENO1 in regulating cell-cell and cell-matrix contacts. We therefore investigated the effect of ENO1 silencing on the modulation of cell morphology, adhesion to matrix substrates, cell invasiveness, and metastatic ability.
  
  
  
  The membrane and cytoskeleton modifications that occurred in ENO1-silenced (shENO1) PDA cells were investigated by a combination of confocal microscopy and atomic force microscopy (AFM). The effect of ENO1 silencing was then evaluated by phenotypic and functional experiments to identify the role of ENO1 in adhesion, migration, and invasion, as well as in senescence and apoptosis. The experimental results were then validated in a mouse model.
  
  
  
  We observed a significant increase in the roughness of the cell membrane due to ENO1 silencing, a feature associated with an impaired ability to migrate and invade, along with a significant downregulation of proteins involved in cell-cell and cell-matrix adhesion, including alpha v/beta 3 integrin in shENO1 PDA cells. These changes impaired the ability of shENO1 cells to adhere to Collagen I and IV and Fibronectin and caused an increase in RGD-independent adhesion to vitronectin (VN) via urokinase plasminogen activator receptor (uPAR). Binding of uPAR to VN triggers integrin-mediated signals, which result in ERK1-2 and RAC activation, accumulation of ROS, and senescence. In shENO1 cancer cells, the use of an anti-uPAR antibody caused significant reduction of ROS production and senescence. Overall, a decrease of in vitro and in vivo cell migration and invasion of shENO1 PDA cells was observed.
  
  
  
  These data demonstrate that ENO1 promotes PDA survival, migration, and metastasis through cooperation with integrins and uPAR.
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Moitza
authors.initials: M
authors.lastname: Principe
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Borgoni
authors.affiliation: Dipartimento di Matematica e Fisica "Ennio De Giorgi", Università del Salento, Lecce, Italy.|Euromediterranean Center for Nanomaterial Modelling and Technology (ECMT) of the Consiglio Nazionale delle Ricerche, Istituto Nanoscienze, Lecce, Italy.
authors.forename: Mariafrancesca
authors.initials: M
authors.lastname: Cascione
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Michelle Samuel
authors.initials: MS
authors.lastname: Chattaragada
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Sammy
authors.initials: S
authors.lastname: Ferri-Borgogno
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Capello
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Bulfamante
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Molecular Biotechnology Center, University of Turin, Turin, Italy.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Chapelle
authors.affiliation: Regina Elena National Cancer Institute, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Di Modugno
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Molecular Biotechnology Center, University of Turin, Turin, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Defilippi
authors.affiliation: Regina Elena National Cancer Institute, Rome, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Nisticò
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy.|Molecular Biotechnology Center, University of Turin, Turin, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cappello
authors.affiliation: Department of Oncology, University of Turin, Turin, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Riganti
authors.affiliation: CNR Nantotec-Istituto di Nanotecnologia, Polo di Nanotecnologia c/o Campus Ecoteckne, Lecce, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Leporatti
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy. franco.novelli@unito.it.|Center for Experimental Research and Medical Studies (CeRMS), Azienda Universitaria Ospedaliera Città della Salute e della Scienza di Torino, Via Santena 5, 10126, Turin, Italy. franco.novelli@unito.it.|Molecular Biotechnology Center, University of Turin, Turin, Italy. franco.novelli@unito.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Novelli
mesh_terms: D000818:Animals; D017209:Apoptosis; D014408:Biomarkers, Tumor / Q000235:genetics / Q000502:physiology*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D048430:Cell Shape; D016922:Cellular Senescence; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D015870:Gene Expression; D020868:Gene Silencing; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000235:genetics* / Q000378:metabolism; D016023:Integrins / Q000378:metabolism / Q000502:physiology; D051379:Mice; D009361:Neoplasm Invasiveness / Q000473:pathology*; D009362:Neoplasm Metastasis / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000502:physiology*; D055293:Receptors, Urokinase Plasminogen Activator / Q000378:metabolism / Q000502:physiology; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D039302:Integrin alphaVbeta3; D016023:Integrins; D055293:Receptors, Urokinase Plasminogen Activator; D025521:Tumor Suppressor Proteins; C095157:ENO1 protein, human; D010751:Phosphopyruvate Hydratase
keywords: Atomic force microscopy; ENO1; Integrin; Invasion; Pancreatic cancer