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CD99 suppresses osteosarcoma cell migration through inhibition of ROCK2 activity

Oncogene volume 33, pages 1912–1921 (2014)Cite this article

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Abstract

CD99, a transmembrane protein encoded by MIC2 gene is involved in multiple cellular events including cell adhesion and migration, apoptosis, cell differentiation and regulation of protein trafficking either in physiological or pathological conditions. In osteosarcoma, CD99 is expressed at low levels and functions as a tumour suppressor. The full-length protein (CD99wt) and the short-form harbouring a deletion in the intracytoplasmic domain (CD99sh) have been associated with distinct functional outcomes with respect to tumour malignancy. In this study, we especially evaluated modulation of cell–cell contacts, reorganisation of the actin cytoskeleton and modulation of signalling pathways by comparing osteosarcoma cells characterised by different metastasis capabilities and CD99 expression, to identify molecular mechanisms responsible for metastasis. Our data indicate that forced expression of CD99wt induces recruitment of N-cadherin and β-catenin to adherens junctions. In addition, transfection of CD99wt inhibits the expression of several molecules crucial to the remodelling of the actin cytoskeleton, such as ACTR2, ARPC1A, Rho-associated, coiled–coil containing protein kinase 2 (ROCK2) as well as ezrin, an ezrin/radixin/moesin family member that has been clearly associated with tumour progression and metastatic spread in osteosarcoma. Functional studies point to ROCK2 as a crucial intracellular mediator regulating osteosarcoma migration. By maintaining c-Src in an inactive conformation, CD99wt inhibits ROCK2 signalling and this leads to ezrin decrease at cell membrane while N-cadherin and β-catenin translocate to the plasma membrane and function as main molecular bridges for actin cytoskeleton. Taken together, we propose that the re-expression of CD99wt, which is generally present in osteoblasts but lost in osteosarcoma, through inhibition of c-Src and ROCK2 activity, manages to increase contact strength and reactivate stop-migration signals that counteract the otherwise dominant promigratory action of ezrin in osteosarcoma cells.

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Acknowledgements

We are in debt to Cristina Ghinelli for editing the manuscript and to Anming Meng, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China, for kindly providing the plasmid pCMV5-HA3-ROCK2. This work was supported by the Italian Association for Cancer Research (AIRC; IG10452 to K Scotlandi), the Liddy Shriver Sarcoma Initiative (international grant to K Scotlandi) and Ricerca Fondamentale Orientata (RFO 2010 to C Zucchini). Rosa Simona Pinca is a recipient of a fellowship from the Associazione Onlus ‘il Pensatore: Matteo Amitrano’ and ‘Liberi di Vivere Luca Righi.’

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Authors and Affiliations

  1. Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy

    C Zucchini, P De Sanctis, P-L Lollini & L Valvassori

  2. CRS Development of Biomolecular Therapies, Bologna, Italy

    M C Manara, R S Pinca, M Sciandra & K Scotlandi

  3. Experimental Oncology Laboratory, Istituto Ortopedico Rizzoli, Bologna, Italy

    M C Manara, R S Pinca, C Guerzoni, P Picci & K Scotlandi

  4. RIT Department, PROMETEO Laboratory, STB, Istituto Ortopedico Rizzoli, Bologna, Italy

    C Guerzoni & K Scotlandi

  5. Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy

    G Cenacchi

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  1. C Zucchini
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Zucchini, C., Manara, M., Pinca, R. et al. CD99 suppresses osteosarcoma cell migration through inhibition of ROCK2 activity. Oncogene 33, 1912–1921 (2014). https://doi.org/10.1038/onc.2013.152

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