Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation

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 MarchisioMaria V. Carriero, Dolores Di Vizio, Alfredo Budillon

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Article number317
JournalJournal of Experimental and Clinical Cancer Research
Volume38
DOIs
Publication statusPublished - Jan 1 2019

Fingerprint

Integrin alphaV
Prostatic Neoplasms
Integrins
Heterografts
Blister
Neoplasms
Mevalonic Acid
Blocking Antibodies
Neoplasm Grading
Cytoskeleton
Centrifugation
Nude Mice
Proteomics
Fluorescent Antibody Technique
Flow Cytometry
Lymph Nodes
Western Blotting
Immunohistochemistry
Staining and Labeling
Technology

Keywords

  • AKT
  • Alpha-V integrin
  • Extracellular vesicles
  • Oncosomes
  • Prostate cancer

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

@article{74115126ff0a49a4a01f5db59e3ee4cf,
title = "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 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.",
keywords = "AKT, Alpha-V integrin, Extracellular vesicles, Oncosomes, Prostate cancer",
author = "Chiara Ciardiello and Alessandra Leone and Paola Lanuti and Roca, {Maria S.} and Tania Moccia and Minciacchi, {Valentina R.} and Michele Minopoli and Vincenzo Gigantino and {De Cecio}, Rossella and Massimo Rippa and Lucia Petti and Francesca Capone and Carlo Vitagliano and Milone, {Maria R.} and Biagio Pucci and Rita Lombardi and Federica Iannelli and {Di Gennaro}, Elena and Francesca Bruzzese and Marco Marchisio and Carriero, {Maria V.} and {Di Vizio}, Dolores and Alfredo Budillon",
year = "2019",
month = "1",
day = "1",
doi = "10.1186/s13046-019-1317-6",
language = "English",
volume = "38",
journal = "Journal of Experimental and Clinical Cancer Research",
issn = "0392-9078",
publisher = "BioMed Central Ltd.",

}

TY - JOUR

T1 - Large oncosomes overexpressing integrin alpha-V promote prostate cancer adhesion and invasion via AKT activation

AU - Ciardiello, Chiara

AU - Leone, Alessandra

AU - Lanuti, Paola

AU - Roca, Maria S.

AU - Moccia, Tania

AU - Minciacchi, Valentina R.

AU - Minopoli, Michele

AU - Gigantino, Vincenzo

AU - De Cecio, Rossella

AU - Rippa, Massimo

AU - Petti, Lucia

AU - Capone, Francesca

AU - Vitagliano, Carlo

AU - Milone, Maria R.

AU - Pucci, Biagio

AU - Lombardi, Rita

AU - Iannelli, Federica

AU - Di Gennaro, Elena

AU - Bruzzese, Francesca

AU - Marchisio, Marco

AU - Carriero, Maria V.

AU - Di Vizio, Dolores

AU - Budillon, Alfredo

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

KW - AKT

KW - Alpha-V integrin

KW - Extracellular vesicles

KW - Oncosomes

KW - Prostate cancer

UR - http://www.scopus.com/inward/record.url?scp=85070093334&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85070093334&partnerID=8YFLogxK

U2 - 10.1186/s13046-019-1317-6

DO - 10.1186/s13046-019-1317-6

M3 - Article

C2 - 31319863

AN - SCOPUS:85070093334

VL - 38

JO - Journal of Experimental and Clinical Cancer Research

JF - Journal of Experimental and Clinical Cancer Research

SN - 0392-9078

M1 - 317

ER -