Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells

BACKGROUND: Glioblastoma is one of the most aggressive cancers of the brain. Malignant traits of glioblastoma cells include elevated migration, proliferation and survival capabilities. Galectins are unconventionally secreted glycan-binding proteins that modulate processes of cell adhesion, migration...

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Autores principales:	Metz,Claudia, Döger,Remziye, Riquelme,Elizabeth, Cortés,Priscilla, Holmes,Christopher, Shaughnessy,Ronan, Oyanadel,Claudia, Grabowski,Catalina, González,Alfonso, Soza,Andrea
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2016
Materias:	Galectin-8 Glioblastoma Cancer Cell cycle Apoptosis Proliferation Migration
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602016000100033
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spelling	oai:scielo:S0716-976020160001000332016-12-01Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cellsMetz,ClaudiaDöger,RemziyeRiquelme,ElizabethCortés,PriscillaHolmes,ChristopherShaughnessy,RonanOyanadel,ClaudiaGrabowski,CatalinaGonzález,AlfonsoSoza,Andrea Galectin-8 Glioblastoma Cancer Cell cycle Apoptosis Proliferation Migration BACKGROUND: Glioblastoma is one of the most aggressive cancers of the brain. Malignant traits of glioblastoma cells include elevated migration, proliferation and survival capabilities. Galectins are unconventionally secreted glycan-binding proteins that modulate processes of cell adhesion, migration, proliferation and apoptosis by interacting with beta-galactosides of cell surface glycoproteins and the extracellular matrix. Galectin-8 is one of the galectins highly expressed in glioblastoma cells. It has a unique selectivity for terminally sialylated glycans recently found enhanced in these highly malignant cells. A previous study in glioblastoma cell lines reported that Gal-8 coating a plastic surface stimulates two-dimensional motility. Because in other cells Gal-8 arrests proliferation and induces apoptosis, here we extend its study by analyzing all of these processes in a U87 glioblastoma cell mode.l METHODS: We used immunoblot and RT-PCR for Gal-8 expression analysis, recombinant Gal-8 produced in a bacteria system for Gal-8 treatment of the cells, and shRNA in lentivirus transduction for Gal-8 silencing. Cell migration as assessed in transwell filters. Cell proliferation, cell cycle and apoptosis were analyzed by FACS. RESULTS: Gal-8 as a soluble stimulus triggered chemotactic migration of U87 cells across the polycarbonate filter of transwell chambers, almost as intensively as fetal bovine serum. Unexpectedly, Gal-8 also enhanced U87 cell growth. Co-incubation of Gal-8 with lactose, which blocks galectin-glycan interactions, abrogated both effects. Immunoblot showed Gal-8 in conditioned media reflecting its secretion. U87 cells transduced with silencing shRNA in a lentiviral vector expressed and secreted 30-40 % of their normal Gal-8 levels. These cells maintained their migratory capabilities, but decreased their proliferation rate and underwent higher levels of apoptosis, as revealed by flow cytometry analysis of cell cycle, CFSE and activated caspase-3 staining. Proliferation seemed to be more sensitive than migration to Gal-8 expression levels. CONCLUSIONS: Gal-8, either secreted or exogenously enriched in the media, and acting through extracellular glycan interactions, constitutes a strong stimulus of directional migration in glioblastoma U87 cells and for the first time emerges as a factor that promotes proliferation and prevents apoptosis in cancerous cells. These properties could potentially contribute to the exaggerated malignancy of glioblastoma cells.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.49 20162016-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602016000100033en10.1186/s40659-016-0091-6
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Galectin-8 Glioblastoma Cancer Cell cycle Apoptosis Proliferation Migration
spellingShingle	Galectin-8 Glioblastoma Cancer Cell cycle Apoptosis Proliferation Migration Metz,Claudia Döger,Remziye Riquelme,Elizabeth Cortés,Priscilla Holmes,Christopher Shaughnessy,Ronan Oyanadel,Claudia Grabowski,Catalina González,Alfonso Soza,Andrea Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
description	BACKGROUND: Glioblastoma is one of the most aggressive cancers of the brain. Malignant traits of glioblastoma cells include elevated migration, proliferation and survival capabilities. Galectins are unconventionally secreted glycan-binding proteins that modulate processes of cell adhesion, migration, proliferation and apoptosis by interacting with beta-galactosides of cell surface glycoproteins and the extracellular matrix. Galectin-8 is one of the galectins highly expressed in glioblastoma cells. It has a unique selectivity for terminally sialylated glycans recently found enhanced in these highly malignant cells. A previous study in glioblastoma cell lines reported that Gal-8 coating a plastic surface stimulates two-dimensional motility. Because in other cells Gal-8 arrests proliferation and induces apoptosis, here we extend its study by analyzing all of these processes in a U87 glioblastoma cell mode.l METHODS: We used immunoblot and RT-PCR for Gal-8 expression analysis, recombinant Gal-8 produced in a bacteria system for Gal-8 treatment of the cells, and shRNA in lentivirus transduction for Gal-8 silencing. Cell migration as assessed in transwell filters. Cell proliferation, cell cycle and apoptosis were analyzed by FACS. RESULTS: Gal-8 as a soluble stimulus triggered chemotactic migration of U87 cells across the polycarbonate filter of transwell chambers, almost as intensively as fetal bovine serum. Unexpectedly, Gal-8 also enhanced U87 cell growth. Co-incubation of Gal-8 with lactose, which blocks galectin-glycan interactions, abrogated both effects. Immunoblot showed Gal-8 in conditioned media reflecting its secretion. U87 cells transduced with silencing shRNA in a lentiviral vector expressed and secreted 30-40 % of their normal Gal-8 levels. These cells maintained their migratory capabilities, but decreased their proliferation rate and underwent higher levels of apoptosis, as revealed by flow cytometry analysis of cell cycle, CFSE and activated caspase-3 staining. Proliferation seemed to be more sensitive than migration to Gal-8 expression levels. CONCLUSIONS: Gal-8, either secreted or exogenously enriched in the media, and acting through extracellular glycan interactions, constitutes a strong stimulus of directional migration in glioblastoma U87 cells and for the first time emerges as a factor that promotes proliferation and prevents apoptosis in cancerous cells. These properties could potentially contribute to the exaggerated malignancy of glioblastoma cells.
author	Metz,Claudia Döger,Remziye Riquelme,Elizabeth Cortés,Priscilla Holmes,Christopher Shaughnessy,Ronan Oyanadel,Claudia Grabowski,Catalina González,Alfonso Soza,Andrea
author_facet	Metz,Claudia Döger,Remziye Riquelme,Elizabeth Cortés,Priscilla Holmes,Christopher Shaughnessy,Ronan Oyanadel,Claudia Grabowski,Catalina González,Alfonso Soza,Andrea
author_sort	Metz,Claudia
title	Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
title_short	Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
title_full	Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
title_fullStr	Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
title_full_unstemmed	Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells
title_sort	galectin-8 promotes migration and proliferation and prevents apoptosis in u87 glioblastoma cells
publisher	Sociedad de Biología de Chile
publishDate	2016
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602016000100033
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Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells

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