ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity
Abstract Background The treatment of metastatic osteosarcoma (OS) remains a challenge for oncologists, and novel therapeutic strategies are urgently needed. An understanding of the pathways that regulate OS dissemination is required for the design of novel treatment approaches. We recently identifie...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13046-019-1506-3 |
| _version_ | 1818616252765044736 |
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| author | Cinzia Zucchini Maria Cristina Manara Camilla Cristalli Marianna Carrabotta Sara Greco Rosa Simona Pinca Cristina Ferrari Lorena Landuzzi Michela Pasello Pier-Luigi Lollini Marco Gambarotti Davide Maria Donati Katia Scotlandi |
| author_facet | Cinzia Zucchini Maria Cristina Manara Camilla Cristalli Marianna Carrabotta Sara Greco Rosa Simona Pinca Cristina Ferrari Lorena Landuzzi Michela Pasello Pier-Luigi Lollini Marco Gambarotti Davide Maria Donati Katia Scotlandi |
| author_sort | Cinzia Zucchini |
| collection | DOAJ |
| description | Abstract Background The treatment of metastatic osteosarcoma (OS) remains a challenge for oncologists, and novel therapeutic strategies are urgently needed. An understanding of the pathways that regulate OS dissemination is required for the design of novel treatment approaches. We recently identified Rho-associated coiled-coil containing protein kinase 2 (ROCK2) as a crucial driver of OS cell migration. In this study, we explored the impact of ROCK2 disruption on the metastatic capabilities of OS cells and analyzed its functional relationship with Yes-associated protein-1 (YAP), the main transcriptional mediator of mechanotransduction signaling. Methods The effects of ROCK2 depletion on metastasis were studied in NOD Scid gamma (NSG) mice injected with U-2OS cells in which ROCK2 expression had been stably silenced. Functional studies were performed in vitro in human U-2OS cells and in three novel cell lines derived from patient-derived xenografts (PDXs) by using standard methods to evaluate malignancy parameters and signaling transduction. The nuclear immunostaining of YAP and the evaluation of its downstream targets Cysteine Rich Angiogenic Inducer 6, Connective Tissue Growth Factor and Cyclin D1 by quantitative PCR were performed to analyze YAP activity. The effect of the expression and activity of ROCK2 and YAP on tumor progression was analyzed in 175 OS primary tumors. Results The silencing of ROCK2 markedly reduced tumor growth and completely abolished the metastatic ability of U-2OS cells. The depletion of ROCK2, either by pharmacological inhibition or silencing, induced a dose- and time-dependent reduction in the nuclear expression and transcriptional activity of YAP. The nuclear expression of YAP was observed in 80/175 (46%) tumor samples and was significantly correlated with worse patient prognosis and a higher likelihood of metastasis and death. The use of verteporfin, a molecule that specifically inhibits the TEAD–YAP association, remarkably impaired the growth and migration of OS cells in vitro. Moreover to inhibiting YAP activity, our findings indicate that verteporfin also affects the ROCK2 protein and its functions. Conclusions We describe the functional connection between ROCK2 and YAP in the regulation of OS cell migration and metastasis formation. These data provide support for the use of verteporfin as a possible therapeutic option to prevent OS cell dissemination. |
| first_indexed | 2024-12-16T16:46:51Z |
| format | Article |
| id | doaj.art-5cd4ab50637442cbb9538299c71ce2ac |
| institution | Directory Open Access Journal |
| issn | 1756-9966 |
| language | English |
| last_indexed | 2024-12-16T16:46:51Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj.art-5cd4ab50637442cbb9538299c71ce2ac2022-12-21T22:24:10ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662019-12-0138111410.1186/s13046-019-1506-3ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activityCinzia Zucchini0Maria Cristina Manara1Camilla Cristalli2Marianna Carrabotta3Sara Greco4Rosa Simona Pinca5Cristina Ferrari6Lorena Landuzzi7Michela Pasello8Pier-Luigi Lollini9Marco Gambarotti10Davide Maria Donati11Katia Scotlandi12Department of Experimental, Diagnostic and Specialty Medicine, (DIMES), University of BolognaExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliDepartment of Experimental, Diagnostic and Specialty Medicine, (DIMES), University of BolognaDepartment of Pathology, IRCCS Istituto Ortopedico RizzoliClinica Ortopedica III, IRCCS Istituto Ortopedico RizzoliExperimental Oncology Laboratory, IRCCS Istituto Ortopedico RizzoliAbstract Background The treatment of metastatic osteosarcoma (OS) remains a challenge for oncologists, and novel therapeutic strategies are urgently needed. An understanding of the pathways that regulate OS dissemination is required for the design of novel treatment approaches. We recently identified Rho-associated coiled-coil containing protein kinase 2 (ROCK2) as a crucial driver of OS cell migration. In this study, we explored the impact of ROCK2 disruption on the metastatic capabilities of OS cells and analyzed its functional relationship with Yes-associated protein-1 (YAP), the main transcriptional mediator of mechanotransduction signaling. Methods The effects of ROCK2 depletion on metastasis were studied in NOD Scid gamma (NSG) mice injected with U-2OS cells in which ROCK2 expression had been stably silenced. Functional studies were performed in vitro in human U-2OS cells and in three novel cell lines derived from patient-derived xenografts (PDXs) by using standard methods to evaluate malignancy parameters and signaling transduction. The nuclear immunostaining of YAP and the evaluation of its downstream targets Cysteine Rich Angiogenic Inducer 6, Connective Tissue Growth Factor and Cyclin D1 by quantitative PCR were performed to analyze YAP activity. The effect of the expression and activity of ROCK2 and YAP on tumor progression was analyzed in 175 OS primary tumors. Results The silencing of ROCK2 markedly reduced tumor growth and completely abolished the metastatic ability of U-2OS cells. The depletion of ROCK2, either by pharmacological inhibition or silencing, induced a dose- and time-dependent reduction in the nuclear expression and transcriptional activity of YAP. The nuclear expression of YAP was observed in 80/175 (46%) tumor samples and was significantly correlated with worse patient prognosis and a higher likelihood of metastasis and death. The use of verteporfin, a molecule that specifically inhibits the TEAD–YAP association, remarkably impaired the growth and migration of OS cells in vitro. Moreover to inhibiting YAP activity, our findings indicate that verteporfin also affects the ROCK2 protein and its functions. Conclusions We describe the functional connection between ROCK2 and YAP in the regulation of OS cell migration and metastasis formation. These data provide support for the use of verteporfin as a possible therapeutic option to prevent OS cell dissemination.https://doi.org/10.1186/s13046-019-1506-3OsteosarcomaROCK2YAPVerteporfinMetastasis |
| spellingShingle | Cinzia Zucchini Maria Cristina Manara Camilla Cristalli Marianna Carrabotta Sara Greco Rosa Simona Pinca Cristina Ferrari Lorena Landuzzi Michela Pasello Pier-Luigi Lollini Marco Gambarotti Davide Maria Donati Katia Scotlandi ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity Journal of Experimental & Clinical Cancer Research Osteosarcoma ROCK2 YAP Verteporfin Metastasis |
| title | ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity |
| title_full | ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity |
| title_fullStr | ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity |
| title_full_unstemmed | ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity |
| title_short | ROCK2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of YAP activity |
| title_sort | rock2 deprivation leads to the inhibition of tumor growth and metastatic potential in osteosarcoma cells through the modulation of yap activity |
| topic | Osteosarcoma ROCK2 YAP Verteporfin Metastasis |
| url | https://doi.org/10.1186/s13046-019-1506-3 |
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