Vascular endothelial cells (EC) and several cancer cells express B7h1, the ligand of the ICOS T cell costimulatory molecule2. We have shown that B7h triggering by a soluble form of ICOS (ICOS-Fc) inhibits the adhesion of polymorphonuclear and tumor cell lines to EC and alters dendritic cells behaviour modulating the cytokine secretion, inhibiting the adhesiveness to EC and the migratory response3,4. Since cancer cell migration is crucial for metastasis dissemination, the aim of this work was to evaluate the effect of ICOS-Fc on migration of cancer cells and EC. METHODS: To evaluate the ICOS-Fc effect on cell migration, the EC and the tumor cell lines were seeded in the upper chamber of a Boyden chamber in the presence and absence of ICOS-Fc and allowed to migrate toward the lower chamber containing medium supplemented or not with either VEGF-A or serum. To assess the effect of ICOS-Fc on epithelial to mesenchymal transition (EMT), HepG2 cells were treated with hepatocyte growth factor (HGF)5. To understand whether the treatment with ICOS-Fc modulated expression of phospho focal adhesion kinase (p-FAK) and beta-PIX6 their expression was analyzed by Western Blot. The effect of B7h triggering on metastatization capability in vivo was assessed by injecting NOD-SCID-IL2Rγnull (NSG) mice with CF-PAC1 Luciferase cells or C57BL/6 mice with B16-F10 cells. RESULTS: ICOS-Fc inhibited the migration of EC, and the HT29, HCT116, PC-3, HepG2, JR8, and M14 tumor cell lines, expressing high levels of B7h. Furthermore, ICOS-Fc down-modulated HGF-induced EMT, and the expression of p-FAK and β-Pix in both EC and tumor cell lines. Finally, the treatment with ICOS-Fc inhibited the development of lung metastases in NSG mice and C57BL/6 mice. DISCUSSION: This work has shown that ICOS binding to B7h influences the cancer progression by acting on both EC and tumor cells. CONCLUSIONS: A drug like ICOS-Fc may be a tool in cancer therapy acting on several aspects of tumor progression.
B7h triggering inhibits the migration of tumor cell lines x
DIANZANI, Chiara;OCCHIPINTI, SERGIO;GIOVARELLI, Mirella;Conti, L.;
2014-01-01
Abstract
Vascular endothelial cells (EC) and several cancer cells express B7h1, the ligand of the ICOS T cell costimulatory molecule2. We have shown that B7h triggering by a soluble form of ICOS (ICOS-Fc) inhibits the adhesion of polymorphonuclear and tumor cell lines to EC and alters dendritic cells behaviour modulating the cytokine secretion, inhibiting the adhesiveness to EC and the migratory response3,4. Since cancer cell migration is crucial for metastasis dissemination, the aim of this work was to evaluate the effect of ICOS-Fc on migration of cancer cells and EC. METHODS: To evaluate the ICOS-Fc effect on cell migration, the EC and the tumor cell lines were seeded in the upper chamber of a Boyden chamber in the presence and absence of ICOS-Fc and allowed to migrate toward the lower chamber containing medium supplemented or not with either VEGF-A or serum. To assess the effect of ICOS-Fc on epithelial to mesenchymal transition (EMT), HepG2 cells were treated with hepatocyte growth factor (HGF)5. To understand whether the treatment with ICOS-Fc modulated expression of phospho focal adhesion kinase (p-FAK) and beta-PIX6 their expression was analyzed by Western Blot. The effect of B7h triggering on metastatization capability in vivo was assessed by injecting NOD-SCID-IL2Rγnull (NSG) mice with CF-PAC1 Luciferase cells or C57BL/6 mice with B16-F10 cells. RESULTS: ICOS-Fc inhibited the migration of EC, and the HT29, HCT116, PC-3, HepG2, JR8, and M14 tumor cell lines, expressing high levels of B7h. Furthermore, ICOS-Fc down-modulated HGF-induced EMT, and the expression of p-FAK and β-Pix in both EC and tumor cell lines. Finally, the treatment with ICOS-Fc inhibited the development of lung metastases in NSG mice and C57BL/6 mice. DISCUSSION: This work has shown that ICOS binding to B7h influences the cancer progression by acting on both EC and tumor cells. CONCLUSIONS: A drug like ICOS-Fc may be a tool in cancer therapy acting on several aspects of tumor progression.
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