EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo
Despite a well-established role for the epidermal growth factor receptor (EGFR) in tumorigenesis, EGFR activities and endocytosis in tumors in vivo have not been studied. We labeled endogenous EGFR with GFP by genome-editing of human oral squamous cell carcinoma cells, which were used to examine EGF...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2017-12-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/31993 |
| _version_ | 1828166466282717184 |
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| author | Itziar Pinilla-Macua Alexandre Grassart Umamaheswar Duvvuri Simon C Watkins Alexander Sorkin |
| author_facet | Itziar Pinilla-Macua Alexandre Grassart Umamaheswar Duvvuri Simon C Watkins Alexander Sorkin |
| author_sort | Itziar Pinilla-Macua |
| collection | DOAJ |
| description | Despite a well-established role for the epidermal growth factor receptor (EGFR) in tumorigenesis, EGFR activities and endocytosis in tumors in vivo have not been studied. We labeled endogenous EGFR with GFP by genome-editing of human oral squamous cell carcinoma cells, which were used to examine EGFR-GFP behavior in mouse tumor xenografts in vivo. Intravital multiphoton imaging, confocal imaging of cryosections and biochemical analysis revealed that localization and trafficking patterns, as well as levels of phosphorylation and ubiquitylation of EGFR in tumors in vivo closely resemble patterns and levels observed in the same cells treated with 20–200 pM EGF in vitro. Consistent with the prediction of low ligand concentrations in tumors, EGFR endocytosis was kinase-dependent and blocked by inhibitors of clathrin-mediated internalization; and EGFR activity was insensitive to Cbl overexpression. Collectively, our data suggest that a small pool of active EGFRs is sufficient to drive tumorigenesis by signaling primarily through the Ras-MAPK pathway. |
| first_indexed | 2024-04-12T02:01:50Z |
| format | Article |
| id | doaj.art-e5bff5934d244e2b87f6a671a0cbc5cc |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:01:50Z |
| publishDate | 2017-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-e5bff5934d244e2b87f6a671a0cbc5cc2022-12-22T03:52:39ZengeLife Sciences Publications LtdeLife2050-084X2017-12-01610.7554/eLife.31993EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivoItziar Pinilla-Macua0Alexandre Grassart1Umamaheswar Duvvuri2Simon C Watkins3Alexander Sorkin4https://orcid.org/0000-0002-4446-1920Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDepartment of Molecular Microbial Pathogenesis, Institute Pasteur, Paris, FranceDepartment of Otolaryngology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDepartment of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, United StatesDespite a well-established role for the epidermal growth factor receptor (EGFR) in tumorigenesis, EGFR activities and endocytosis in tumors in vivo have not been studied. We labeled endogenous EGFR with GFP by genome-editing of human oral squamous cell carcinoma cells, which were used to examine EGFR-GFP behavior in mouse tumor xenografts in vivo. Intravital multiphoton imaging, confocal imaging of cryosections and biochemical analysis revealed that localization and trafficking patterns, as well as levels of phosphorylation and ubiquitylation of EGFR in tumors in vivo closely resemble patterns and levels observed in the same cells treated with 20–200 pM EGF in vitro. Consistent with the prediction of low ligand concentrations in tumors, EGFR endocytosis was kinase-dependent and blocked by inhibitors of clathrin-mediated internalization; and EGFR activity was insensitive to Cbl overexpression. Collectively, our data suggest that a small pool of active EGFRs is sufficient to drive tumorigenesis by signaling primarily through the Ras-MAPK pathway.https://elifesciences.org/articles/31993EGF receptortumourendocytosis |
| spellingShingle | Itziar Pinilla-Macua Alexandre Grassart Umamaheswar Duvvuri Simon C Watkins Alexander Sorkin EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo eLife EGF receptor tumour endocytosis |
| title | EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo |
| title_full | EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo |
| title_fullStr | EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo |
| title_full_unstemmed | EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo |
| title_short | EGF receptor signaling, phosphorylation, ubiquitylation and endocytosis in tumors in vivo |
| title_sort | egf receptor signaling phosphorylation ubiquitylation and endocytosis in tumors in vivo |
| topic | EGF receptor tumour endocytosis |
| url | https://elifesciences.org/articles/31993 |
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