Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia
Tissue hypoxia is commonly observed in head and neck squamous cell carcinomas (HNSCCs), resulting in molecular and functional alterations of the tumor cells. The aim of this study was to characterize tumor-derived small extracellular vesicles (sEVs) released under hypoxic vs. normoxic conditions and...
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MDPI AG
2021-08-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/13/16/4176 |
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| author | Alicja Głuszko Mirosław J. Szczepański Theresa L. Whiteside Torsten E. Reichert Jacek Siewiera Nils Ludwig |
| author_facet | Alicja Głuszko Mirosław J. Szczepański Theresa L. Whiteside Torsten E. Reichert Jacek Siewiera Nils Ludwig |
| author_sort | Alicja Głuszko |
| collection | DOAJ |
| description | Tissue hypoxia is commonly observed in head and neck squamous cell carcinomas (HNSCCs), resulting in molecular and functional alterations of the tumor cells. The aim of this study was to characterize tumor-derived small extracellular vesicles (sEVs) released under hypoxic vs. normoxic conditions and analyze their proteomic content. HNSCC cells (FaDu, PCI-30, SCC-25) and HaCaT keratinocytes were cultured in 21, 10, 5, and 1% O<sub>2</sub>. sEVs were isolated from supernatants using size exclusion chromatography (SEC) and characterized by nanoparticle tracking analysis, electron microscopy, immunoblotting, and high-resolution mass spectrometry. Isolated sEVs ranged in size from 125–135 nm and contained CD63 and CD9 but not Grp94. sEVs reflected the hypoxic profile of HNSCC parent cells: about 15% of the total detected proteins were unique for hypoxic cells. Hypoxic sEVs expressed a common signature of seven hypoxia-related proteins (KT33B, DYSF, STON2, MLX, LIPA3, NEK5, P12L1) and were enriched in pro-angiogenic proteins. Protein profiles of sEVs reflected the degree of tumor hypoxia and could serve as potential sEV-based biomarkers for hypoxic conditions. Adaptation of HNSCC cells to hypoxia is associated with increased release of sEVs, which are enriched in a unique protein profile. Thus, tumor-derived sEVs can potentially be useful for evaluating levels of hypoxia in HNSCC. |
| first_indexed | 2024-03-10T08:57:02Z |
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| id | doaj.art-fa9eb61eb7b040f4b4fbeaf374f40664 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T08:57:02Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-fa9eb61eb7b040f4b4fbeaf374f406642023-11-22T07:04:55ZengMDPI AGCancers2072-66942021-08-011316417610.3390/cancers13164176Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor HypoxiaAlicja Głuszko0Mirosław J. Szczepański1Theresa L. Whiteside2Torsten E. Reichert3Jacek Siewiera4Nils Ludwig5Chair and Department of Biochemistry, Medical University of Warsaw, 1 Banacha St., 02-097 Warsaw, PolandChair and Department of Biochemistry, Medical University of Warsaw, 1 Banacha St., 02-097 Warsaw, PolandDepartment of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USADepartment of Oral and Maxillofacial Surgery, University Hospital Regensburg, 93053 Regensburg, GermanyDepartment of Hyperbaric Medicine, Military Institute of Medicine, 04-141 Warsaw, PolandDepartment of Oral and Maxillofacial Surgery, University Hospital Regensburg, 93053 Regensburg, GermanyTissue hypoxia is commonly observed in head and neck squamous cell carcinomas (HNSCCs), resulting in molecular and functional alterations of the tumor cells. The aim of this study was to characterize tumor-derived small extracellular vesicles (sEVs) released under hypoxic vs. normoxic conditions and analyze their proteomic content. HNSCC cells (FaDu, PCI-30, SCC-25) and HaCaT keratinocytes were cultured in 21, 10, 5, and 1% O<sub>2</sub>. sEVs were isolated from supernatants using size exclusion chromatography (SEC) and characterized by nanoparticle tracking analysis, electron microscopy, immunoblotting, and high-resolution mass spectrometry. Isolated sEVs ranged in size from 125–135 nm and contained CD63 and CD9 but not Grp94. sEVs reflected the hypoxic profile of HNSCC parent cells: about 15% of the total detected proteins were unique for hypoxic cells. Hypoxic sEVs expressed a common signature of seven hypoxia-related proteins (KT33B, DYSF, STON2, MLX, LIPA3, NEK5, P12L1) and were enriched in pro-angiogenic proteins. Protein profiles of sEVs reflected the degree of tumor hypoxia and could serve as potential sEV-based biomarkers for hypoxic conditions. Adaptation of HNSCC cells to hypoxia is associated with increased release of sEVs, which are enriched in a unique protein profile. Thus, tumor-derived sEVs can potentially be useful for evaluating levels of hypoxia in HNSCC.https://www.mdpi.com/2072-6694/13/16/4176small extracellular vesiclesexosomeshypoxiaproteomicsHNSCC |
| spellingShingle | Alicja Głuszko Mirosław J. Szczepański Theresa L. Whiteside Torsten E. Reichert Jacek Siewiera Nils Ludwig Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia Cancers small extracellular vesicles exosomes hypoxia proteomics HNSCC |
| title | Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia |
| title_full | Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia |
| title_fullStr | Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia |
| title_full_unstemmed | Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia |
| title_short | Small Extracellular Vesicles from Head and Neck Squamous Cell Carcinoma Cells Carry a Proteomic Signature for Tumor Hypoxia |
| title_sort | small extracellular vesicles from head and neck squamous cell carcinoma cells carry a proteomic signature for tumor hypoxia |
| topic | small extracellular vesicles exosomes hypoxia proteomics HNSCC |
| url | https://www.mdpi.com/2072-6694/13/16/4176 |
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