The Blockade of plasminogen-alpha-enolase interaction inhibits THE invasion OF PANCREATIC CANCER CELLS

Patrizia Ceruti, Michelle Samuel Chattaragada, Paola Cappello, Moitza Principe, Jorg Hamm, Simona Rolla, Franco Novelli

Center for Experimental Research and Medical Studies(CeRMS), San Giovanni Battista Hospital and Department of Medicine and Experimental Oncology,UniversityofTurin,Turin,Italy

 Pancreatic Ductal Adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by rapid progression, invasiveness and resistance to treatment. In our laboratory it has been demonstrated that most of PDAC patients have circulating antibodies against the glycolytic enzyme alpha-enolase. Alpha-enolase is not only a cytoplasmic enzyme but it is also expressed on cell surface where it acts as a plasminogen receptor. By promoting plasminogen activation into plasmin, a serine-protease involved in the extracellular matrix degradation, alpha- enolase could play a crucial role in cell invasion and metastatization. By cytofluorimetry we observed that alpha-enolase is expressed on most of PDAC cell lines. The aim of this study was to investigate its role in PDAC cell invasion. We found that treatment of the PDAC cell line CF-PAC-1 with a mouse monoclonal antibody against human alpha-enolase inhibited plasminogen-dependent migration through Matrigel, without  influencing in vitro cell growth. To validate these results in vivo, SCID-beige mice were injected in the tail vein with luciferase expressing CF-PAC-1 cells and treated with anti-alpha- enolase monoclonal antibody or irrelevant mouse IgG as control. Mice treated with anti-alpha-enolase antibody displayed a reduced number of tumor mass compared to control IgG-treated mice. These data indicate that alpha-enolase is involved in the PDAC invasion suggesting that interference with enolase-plasminogen interaction could be useful to inhibit this process.

Targeting Cancer Invasion and Metastasis, February 21-24, 2010,Miami Beach,Florida,USA

The Blockade of plasminogen-alpha-enolase interaction inhibits THE invasion OF PANCREATIC CANCER CELLS

Patrizia Ceruti, Michelle Samuel Chattaragada, Paola Cappello, Moitza Principe, Jorg Hamm, Simona Rolla, Franco Novelli

Center for Experimental Research and Medical Studies (CeRMS), San Giovanni Battista Hospital and Department of Medicine and Experimental Oncology,UniversityofTurin,Turin,Italy

Introduction: Pancreatic Ductal Adenocarcinoma (PDAC) is a malignancy characterized by rapid progression, invasiveness and resistance to treatment. It has been demonstrated that 70% of PDAC patients have circulating antibodies against the glycolitic enzyme alpha-enolase and this correlates with a longer time free of disease. Alpha-enolase is also expressed on the surface of some cell types where it acts as a plasminogen receptor. By promoting plasminogen activation into plasmin, a serine-protease involved in the extracellular matrix degradation, alpha-enolase could play a crucial role in cell invasion and metastatization. Objective: The aim of this study was to investigate the alpha-enolase role in PDAC cell invasion. Methods: We analyzed surface alpha-enolase expression on PDAC cell lines by flow-cytometry. To asses its role in invasion we performed an in vitro matrigel assay with the PDAC cell line CF-PAC-1 inpresence of plasminogen and anti-alpha-enolase monoclonal antibody. Cell proliferation was assessed in the same conditions by a MTT test. Moreover, SCID-beige mice were injected in the tail vein with luciferase expressing CF-PAC-1 cells and treated with anti-alpha-enolase or control antibodies. Results: We observed that alpha-enolase is expressed on cell surface of most of PDAC lines and the treatment  with the specific antibody inhibited their plasminogen-dependent invasion, without influencing in vitro cell growth. Moreover, mice treated with the anti-alpha-enolase antibody displayed a reduced number of tumor mass compared to control mice. Conclusion: These data indicate that alpha-enolase is involved in the PDAC invasion suggesting that interference with enolase-plasminogen interaction could be of therapeutic value.

Pancreatology 2010; 10:259-400 42^nd European Pancreatic Club (EPC) Meeting June 16-19, 2010,Stockholm,Sweden

Targeting Cancer Invasion and Metastasis, February 21-24, 2010,Miami Beach,Florida,USA

Pancreatology 2010; 10:259-400 42^nd European Pancreatic Club (EPC) Meeting June 16-19, 2010,Stockholm,Sweden