Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation
Abstract Background Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as “Large Oncosomes” (LO), have been identified in highly migratory and invasive PCa cells. We recently developed...
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| Format: | Article |
| Language: | English |
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BMC
2019-07-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
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| Online Access: | http://link.springer.com/article/10.1186/s13046-019-1317-6 |
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| author | Chiara Ciardiello Alessandra Leone Paola Lanuti Maria S. Roca Tania Moccia Valentina R. Minciacchi Michele Minopoli Vincenzo Gigantino Rossella De Cecio Massimo Rippa Lucia Petti Francesca Capone Carlo Vitagliano Maria R. Milone Biagio Pucci Rita Lombardi Federica Iannelli Elena Di Gennaro Francesca Bruzzese Marco Marchisio Maria V. Carriero Dolores Di Vizio Alfredo Budillon |
| author_facet | Chiara Ciardiello Alessandra Leone Paola Lanuti Maria S. Roca Tania Moccia Valentina R. Minciacchi Michele Minopoli Vincenzo Gigantino Rossella De Cecio Massimo Rippa Lucia Petti Francesca Capone Carlo Vitagliano Maria R. Milone Biagio Pucci Rita Lombardi Federica Iannelli Elena Di Gennaro Francesca Bruzzese Marco Marchisio Maria V. Carriero Dolores Di Vizio Alfredo Budillon |
| author_sort | Chiara Ciardiello |
| collection | DOAJ |
| description | Abstract Background Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as “Large Oncosomes” (LO), have been identified in highly migratory and invasive PCa cells. We recently developed and characterized the DU145R80 subline, selected from parental DU145 cells as resistant to inhibitors of mevalonate pathway. DU145R80 showed different proteomic profile compared to parental DU145 cells, along with altered cytoskeleton dynamics and a more aggressive phenotype. Methods Immunofluorescence staining and western blotting were used to identify blebbing and EVs protein cargo. EVs, purified by gradient ultra-centrifugations, were analyzed by tunable resistive pulse sensing and multi-parametric flow cytometry approach coupled with high-resolution imaging technologies. LO functional effects were tested in vitro by adhesion and invasion assays and in vivo xenograft model in nude mice. Xenograft and patient tumor tissues were analyzed by immunohistochemistry. Results We found spontaneous blebbing and increased shedding of LO from DU145R80 compared to DU145 cells. LO from DU145R80, compared to those from DU145, carried increased amounts of key-molecules involved in PCa progression including integrin alpha V (αV-integrin). By incubating DU145 cells with DU145R80-derived LO we demonstrated that αV-integrin on LO surface was functionally involved in the increased adhesion and invasion of recipient cells, via AKT. Indeed either the pre-incubation of LO with an αV-integrin blocking antibody, or a specific AKT inhibition in recipient cells are able to revert the LO-induced functional effects. Moreover, DU145R80-derived LO also increased DU145 tumor engraftment in a mice model. Finally, we identified αV-integrin positive LO-like structures in tumor xenografts as well as in PCa patient tissues. Increased αV-integrin tumor expression correlated with high Gleason score and lymph node status. Conclusions Overall, this study is the first to demonstrate the critical role of αV-integrin positive LO in PCa aggressive features, adding new insights in biological function of these large EVs and suggesting their potential use as PCa prognostic markers. |
| first_indexed | 2024-12-21T23:12:05Z |
| format | Article |
| id | doaj.art-df0b835cb5874271a1a89640e6d4f5e9 |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-12-21T23:12:05Z |
| publishDate | 2019-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-df0b835cb5874271a1a89640e6d4f5e92022-12-21T18:47:00ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662019-07-0138111610.1186/s13046-019-1317-6Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activationChiara Ciardiello0Alessandra Leone1Paola Lanuti2Maria S. Roca3Tania Moccia4Valentina R. Minciacchi5Michele Minopoli6Vincenzo Gigantino7Rossella De Cecio8Massimo Rippa9Lucia Petti10Francesca Capone11Carlo Vitagliano12Maria R. Milone13Biagio Pucci14Rita Lombardi15Federica Iannelli16Elena Di Gennaro17Francesca Bruzzese18Marco Marchisio19Maria V. Carriero20Dolores Di Vizio21Alfredo Budillon22Experimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleCentre on Aging Sciences and Translational Medicine (Ce.S.I.-Me.T.), University “G.d’Annunzio”Experimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleGeorg-Speyer-Haus Institute for Tumor biology and Experimental TherapyNeoplastic Progression Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascalePathology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascalePathology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleInstitute of Applied Sciences and Intelligent Systems ‘E. Caianiello’ of CNRInstitute of Applied Sciences and Intelligent Systems ‘E. Caianiello’ of CNRExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleCentre on Aging Sciences and Translational Medicine (Ce.S.I.-Me.T.), University “G.d’Annunzio”Neoplastic Progression Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleDepartments of Surgery, Pathology & Lab Medicine, and Biochemical Science, Cedars-Sinai Medical CenterExperimental Pharmacology Unit, Istituto Nazionale Tumori – IRCCS- Fondazione G. PascaleAbstract Background Molecular markers for prostate cancer (PCa) are required to improve the early definition of patient outcomes. Atypically large extracellular vesicles (EVs), referred as “Large Oncosomes” (LO), have been identified in highly migratory and invasive PCa cells. We recently developed and characterized the DU145R80 subline, selected from parental DU145 cells as resistant to inhibitors of mevalonate pathway. DU145R80 showed different proteomic profile compared to parental DU145 cells, along with altered cytoskeleton dynamics and a more aggressive phenotype. Methods Immunofluorescence staining and western blotting were used to identify blebbing and EVs protein cargo. EVs, purified by gradient ultra-centrifugations, were analyzed by tunable resistive pulse sensing and multi-parametric flow cytometry approach coupled with high-resolution imaging technologies. LO functional effects were tested in vitro by adhesion and invasion assays and in vivo xenograft model in nude mice. Xenograft and patient tumor tissues were analyzed by immunohistochemistry. Results We found spontaneous blebbing and increased shedding of LO from DU145R80 compared to DU145 cells. LO from DU145R80, compared to those from DU145, carried increased amounts of key-molecules involved in PCa progression including integrin alpha V (αV-integrin). By incubating DU145 cells with DU145R80-derived LO we demonstrated that αV-integrin on LO surface was functionally involved in the increased adhesion and invasion of recipient cells, via AKT. Indeed either the pre-incubation of LO with an αV-integrin blocking antibody, or a specific AKT inhibition in recipient cells are able to revert the LO-induced functional effects. Moreover, DU145R80-derived LO also increased DU145 tumor engraftment in a mice model. Finally, we identified αV-integrin positive LO-like structures in tumor xenografts as well as in PCa patient tissues. Increased αV-integrin tumor expression correlated with high Gleason score and lymph node status. Conclusions Overall, this study is the first to demonstrate the critical role of αV-integrin positive LO in PCa aggressive features, adding new insights in biological function of these large EVs and suggesting their potential use as PCa prognostic markers.http://link.springer.com/article/10.1186/s13046-019-1317-6Extracellular vesiclesOncosomesProstate cancerAlpha-V integrinAKT |
| spellingShingle | Chiara Ciardiello Alessandra Leone Paola Lanuti Maria S. Roca Tania Moccia Valentina R. Minciacchi Michele Minopoli Vincenzo Gigantino Rossella De Cecio Massimo Rippa Lucia Petti Francesca Capone Carlo Vitagliano Maria R. Milone Biagio Pucci Rita Lombardi Federica Iannelli Elena Di Gennaro Francesca Bruzzese Marco Marchisio Maria V. Carriero Dolores Di Vizio Alfredo Budillon Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation Journal of Experimental & Clinical Cancer Research Extracellular vesicles Oncosomes Prostate cancer Alpha-V integrin AKT |
| title | Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation |
| title_full | Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation |
| title_fullStr | Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation |
| title_full_unstemmed | Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation |
| title_short | Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation |
| title_sort | large oncosomes overexpressing integrin alpha v promote prostate cancer adhesion and invasion via akt activation |
| topic | Extracellular vesicles Oncosomes Prostate cancer Alpha-V integrin AKT |
| url | http://link.springer.com/article/10.1186/s13046-019-1317-6 |
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