Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration
Extracellular vesicles (EVs) released from tumor cells are actively investigated, since molecules therein contained and likely transferred to neighboring cells, supplying them with oncogenic information/functions, may represent cancer biomarkers and/or druggable targets. Here, we characterized by a...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-02-01
|
| Series: | Biomedicines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2227-9059/9/2/146 |
| _version_ | 1797415980224217088 |
|---|---|
| author | Fabrizio Di Giuseppe Marzia Carluccio Mariachiara Zuccarini Patricia Giuliani Lucia Ricci-Vitiani Roberto Pallini Paolo De Sanctis Roberta Di Pietro Renata Ciccarelli Stefania Angelucci |
| author_facet | Fabrizio Di Giuseppe Marzia Carluccio Mariachiara Zuccarini Patricia Giuliani Lucia Ricci-Vitiani Roberto Pallini Paolo De Sanctis Roberta Di Pietro Renata Ciccarelli Stefania Angelucci |
| author_sort | Fabrizio Di Giuseppe |
| collection | DOAJ |
| description | Extracellular vesicles (EVs) released from tumor cells are actively investigated, since molecules therein contained and likely transferred to neighboring cells, supplying them with oncogenic information/functions, may represent cancer biomarkers and/or druggable targets. Here, we characterized by a proteomic point of view two EV subtypes isolated by sequential centrifugal ultrafiltration technique from culture medium of glioblastoma (GBM)-derived stem-like cells (GSCs) obtained from surgical specimens of human GBM, the most aggressive and lethal primary brain tumor. Electron microscopy and western blot analysis distinguished them into microvesicles (MVs) and exosomes (Exos). Two-dimensional electrophoresis followed by MALDI TOF analysis allowed us to identify, besides a common pool, sets of proteins specific for each EV subtypes with peculiar differences in their molecular/biological functions. Such a diversity was confirmed by identification of some top proteins selected in MVs and Exos. They were mainly chaperone or metabolic enzymes in MVs, whereas, in Exos, molecules are involved in cell–matrix adhesion, cell migration/aggressiveness, and chemotherapy resistance. These proteins, identified by EVs from primary GSCs and not GBM cell lines, could be regarded as new possible prognostic markers/druggable targets of the human tumor, although data need to be confirmed in EVs isolated from a greater GSC number. |
| first_indexed | 2024-03-09T05:57:18Z |
| format | Article |
| id | doaj.art-6f07a6cf2bd7443e968f12d17f0acbcf |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T05:57:18Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-6f07a6cf2bd7443e968f12d17f0acbcf2023-12-03T12:12:52ZengMDPI AGBiomedicines2227-90592021-02-019214610.3390/biomedicines9020146Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal UltrafiltrationFabrizio Di Giuseppe0Marzia Carluccio1Mariachiara Zuccarini2Patricia Giuliani3Lucia Ricci-Vitiani4Roberto Pallini5Paolo De Sanctis6Roberta Di Pietro7Renata Ciccarelli8Stefania Angelucci9Department of Innovative Technologies in Medicine and Dentistry, ‘G. d’Annunzio’ University of Chieti-Pescara, Via Vestini 31, 66100 Chieti, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyDepartment of Oncology and Molecular Medicine, Istituto Superiore di Sanità, Via Regina Elena 299, 00161 Rome, ItalyInstitute of Neurosurgery, Università Cattolica del Sacro Cuore, Largo Agostino Gemelli 8, 00168 Rome, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyCenter for Advanced Studies and Technology (CAST), ‘G. d’Annunzio’ University of Chieti-Pescara, Via L Polacchi 13, 66100 Chieti, ItalyDepartment of Innovative Technologies in Medicine and Dentistry, ‘G. d’Annunzio’ University of Chieti-Pescara, Via Vestini 31, 66100 Chieti, ItalyExtracellular vesicles (EVs) released from tumor cells are actively investigated, since molecules therein contained and likely transferred to neighboring cells, supplying them with oncogenic information/functions, may represent cancer biomarkers and/or druggable targets. Here, we characterized by a proteomic point of view two EV subtypes isolated by sequential centrifugal ultrafiltration technique from culture medium of glioblastoma (GBM)-derived stem-like cells (GSCs) obtained from surgical specimens of human GBM, the most aggressive and lethal primary brain tumor. Electron microscopy and western blot analysis distinguished them into microvesicles (MVs) and exosomes (Exos). Two-dimensional electrophoresis followed by MALDI TOF analysis allowed us to identify, besides a common pool, sets of proteins specific for each EV subtypes with peculiar differences in their molecular/biological functions. Such a diversity was confirmed by identification of some top proteins selected in MVs and Exos. They were mainly chaperone or metabolic enzymes in MVs, whereas, in Exos, molecules are involved in cell–matrix adhesion, cell migration/aggressiveness, and chemotherapy resistance. These proteins, identified by EVs from primary GSCs and not GBM cell lines, could be regarded as new possible prognostic markers/druggable targets of the human tumor, although data need to be confirmed in EVs isolated from a greater GSC number.https://www.mdpi.com/2227-9059/9/2/146glioblastoma (GBM)glioblastoma-derived stem-like cells (GSCs)sequential centrifugal ultrafiltration (SCUF)microvesiclesoncosomesexosomes |
| spellingShingle | Fabrizio Di Giuseppe Marzia Carluccio Mariachiara Zuccarini Patricia Giuliani Lucia Ricci-Vitiani Roberto Pallini Paolo De Sanctis Roberta Di Pietro Renata Ciccarelli Stefania Angelucci Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration Biomedicines glioblastoma (GBM) glioblastoma-derived stem-like cells (GSCs) sequential centrifugal ultrafiltration (SCUF) microvesicles oncosomes exosomes |
| title | Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration |
| title_full | Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration |
| title_fullStr | Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration |
| title_full_unstemmed | Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration |
| title_short | Proteomic Characterization of Two Extracellular Vesicle Subtypes Isolated from Human Glioblastoma Stem Cell Secretome by Sequential Centrifugal Ultrafiltration |
| title_sort | proteomic characterization of two extracellular vesicle subtypes isolated from human glioblastoma stem cell secretome by sequential centrifugal ultrafiltration |
| topic | glioblastoma (GBM) glioblastoma-derived stem-like cells (GSCs) sequential centrifugal ultrafiltration (SCUF) microvesicles oncosomes exosomes |
| url | https://www.mdpi.com/2227-9059/9/2/146 |
| work_keys_str_mv | AT fabriziodigiuseppe proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT marziacarluccio proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT mariachiarazuccarini proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT patriciagiuliani proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT luciariccivitiani proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT robertopallini proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT paolodesanctis proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT robertadipietro proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT renataciccarelli proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration AT stefaniaangelucci proteomiccharacterizationoftwoextracellularvesiclesubtypesisolatedfromhumanglioblastomastemcellsecretomebysequentialcentrifugalultrafiltration |