Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells

The Sprouty (SPRY) proteins are evolutionary conserved modulators of receptor tyrosine kinase (RTK) signaling. SPRY2 inhibits fibroblast growth factor (FGF) signaling, whereas it enhances epidermal growth factor (EGF) signaling through inhibition of EGF receptor (EGFR) endocytosis, ubiquitination, a...

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Main Authors: Barbara Hausott, Lena Pircher, Michaela Kind, Jong-Whi Park, Peter Claus, Petra Obexer, Lars Klimaschewski
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/13/23/1967
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author Barbara Hausott
Lena Pircher
Michaela Kind
Jong-Whi Park
Peter Claus
Petra Obexer
Lars Klimaschewski
author_facet Barbara Hausott
Lena Pircher
Michaela Kind
Jong-Whi Park
Peter Claus
Petra Obexer
Lars Klimaschewski
author_sort Barbara Hausott
collection DOAJ
description The Sprouty (SPRY) proteins are evolutionary conserved modulators of receptor tyrosine kinase (RTK) signaling. SPRY2 inhibits fibroblast growth factor (FGF) signaling, whereas it enhances epidermal growth factor (EGF) signaling through inhibition of EGF receptor (EGFR) endocytosis, ubiquitination, and degradation. In this study, we analyzed the effects of SPRY2 on endocytosis and degradation of FGF receptor 1 (FGFR1) using two human glioblastoma (GBM) cell lines with different endogenous SPRY2 levels. SPRY2 overexpression (SPRY2-OE) inhibited clathrin- and caveolae-mediated endocytosis of FGFR1, reduced the number of caveolin-1 vesicles and the uptake of transferrin. Furthermore, FGFR1 protein was decreased by SPRY2-OE, whereas EGFR protein was increased. SPRY2-OE enhanced FGFR1 degradation by increased c-casitas b-lineage lymphoma (c-CBL)-mediated ubiquitination, but it diminished binding of phospholipase Cγ1 (PLCγ1) to FGFR1. Consequently, SPRY2-OE inhibited FGF2-induced activation of PLCγ1, whereas it enhanced EGF-induced PLCγ1 activation. Despite the reduction of FGFR1 protein and the inhibition of FGF signaling, SPRY2-OE increased cell viability, and knockdown of SPRY2 enhanced the sensitivity to cisplatin. These results demonstrate that the inhibitory effect of SPRY2-OE on FGF signaling is at least in part due to the reduction in FGFR1 levels and the decreased binding of PLCγ1 to the receptor.
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spelling doaj-art-7a01ff78ab784b5fb43dd2dd22fafc7d2024-12-13T16:24:10ZengMDPI AGCells2073-44092024-11-011323196710.3390/cells13231967Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma CellsBarbara Hausott0Lena Pircher1Michaela Kind2Jong-Whi Park3Peter Claus4Petra Obexer5Lars Klimaschewski6Institute of Neuroanatomy, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Neuroanatomy, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Neuroanatomy, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Neuroanatomy, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Functional and Applied Anatomy, Hannover Medical School, 30625 Hannover, GermanyDepartment of Pediatrics II, Medical University of Innsbruck, 6020 Innsbruck, AustriaInstitute of Neuroanatomy, Medical University of Innsbruck, 6020 Innsbruck, AustriaThe Sprouty (SPRY) proteins are evolutionary conserved modulators of receptor tyrosine kinase (RTK) signaling. SPRY2 inhibits fibroblast growth factor (FGF) signaling, whereas it enhances epidermal growth factor (EGF) signaling through inhibition of EGF receptor (EGFR) endocytosis, ubiquitination, and degradation. In this study, we analyzed the effects of SPRY2 on endocytosis and degradation of FGF receptor 1 (FGFR1) using two human glioblastoma (GBM) cell lines with different endogenous SPRY2 levels. SPRY2 overexpression (SPRY2-OE) inhibited clathrin- and caveolae-mediated endocytosis of FGFR1, reduced the number of caveolin-1 vesicles and the uptake of transferrin. Furthermore, FGFR1 protein was decreased by SPRY2-OE, whereas EGFR protein was increased. SPRY2-OE enhanced FGFR1 degradation by increased c-casitas b-lineage lymphoma (c-CBL)-mediated ubiquitination, but it diminished binding of phospholipase Cγ1 (PLCγ1) to FGFR1. Consequently, SPRY2-OE inhibited FGF2-induced activation of PLCγ1, whereas it enhanced EGF-induced PLCγ1 activation. Despite the reduction of FGFR1 protein and the inhibition of FGF signaling, SPRY2-OE increased cell viability, and knockdown of SPRY2 enhanced the sensitivity to cisplatin. These results demonstrate that the inhibitory effect of SPRY2-OE on FGF signaling is at least in part due to the reduction in FGFR1 levels and the decreased binding of PLCγ1 to the receptor.https://www.mdpi.com/2073-4409/13/23/1967caveolin-1c-casitas b-lineage lymphomaclathrinextracellular signal-regulated kinasephospholipase Cγ1ubiquitin
spellingShingle Barbara Hausott
Lena Pircher
Michaela Kind
Jong-Whi Park
Peter Claus
Petra Obexer
Lars Klimaschewski
Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
Cells
caveolin-1
c-casitas b-lineage lymphoma
clathrin
extracellular signal-regulated kinase
phospholipase Cγ1
ubiquitin
title Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
title_full Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
title_fullStr Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
title_full_unstemmed Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
title_short Sprouty2 Regulates Endocytosis and Degradation of Fibroblast Growth Factor Receptor 1 in Glioblastoma Cells
title_sort sprouty2 regulates endocytosis and degradation of fibroblast growth factor receptor 1 in glioblastoma cells
topic caveolin-1
c-casitas b-lineage lymphoma
clathrin
extracellular signal-regulated kinase
phospholipase Cγ1
ubiquitin
url https://www.mdpi.com/2073-4409/13/23/1967
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