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CD82 suppresses CD44 alternative splicing-dependent melanoma metastasis by mediating U2AF2 ubiquitination and degradation

Oncogene volume 35, pages 5056–5069 (2016)Cite this article

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Abstract

Melanoma is one of the most lethal forms of skin cancer because of its early metastatic spread. The variant form of CD44 (CD44v), a cell surface glycoprotein, is highly expressed on metastatic melanoma. The mechanisms of regulation of CD44 alternative splicing in melanoma and its pathogenic contributions are so far poorly understood. Here, we investigated the expression level of CD44 in a large set of melanocytic lesions at different stages. We found that the expression of CD44v8–10 and a splicing factor, U2AF2, is significantly increased during melanoma progression, whereas CD82/KAI1, a tetraspanin family of tumor suppressor, is reduced in metastatic melanoma. CD44v8–10 and U2AF2 expression levels, which are negatively correlated with CD82 levels, are markedly elevated in primary melanoma compared with dysplastic nevi and further increased in metastatic melanoma. We also showed that patients with higher CD44v8–10 and U2AF2 expression levels tended to have shorter survival. By using both in vivo and in vitro assays, we demonstrated that CD82 inhibits the production of CD44v8–10 on melanoma. Mechanistically, U2AF2 is a downstream target of CD82 and in malignant melanoma facilitates CD44v8–10 alternative splicing. U2AF2-mediated CD44 isoform switch is required for melanoma migration in vitro and lung and liver metastasis in vivo. Notably, overexpression of CD82 suppresses U2AF2 activity by inducing U2AF2 ubiquitination. In addition, our data suggested that enhancement of melanoma migration by U2AF2-dependent CD44v8–10 splicing is mediated by Src/focal adhesion kinase/RhoA activation and formation of stress fibers, as well as CD44-E-selectin binding reinforcement. These findings uncovered a hitherto unappreciated function of CD82 in severing the linkage between U2AF2-mediated CD44 alternative splicing and cancer aggressiveness, with potential prognostic and therapeutic implications in melanoma.

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Acknowledgements

We thank Go J Yoshida (Tokyo Medical and Dental University) for helpful discussion. This study was funded by Fundamental Research Funds for the Central Universities (XDJK2014C176 to PZ, XDJK2012B010 to HFZ, XDJK2015C154 to QR and XDJK2015C155 to FS), the Start-up Foundation of Southwest University (SWU114017 to PZ), National Natural Science Foundation of China (NSFC) (NSFC-81402393 to PZ, NSFC-81572678 to PZ, NSFC-81272433 to GTL, NSFC-81472732 to GTL and NSFC-81073084 to HFZ) and National Science Foundation grants (CBET-0729091 to CD).

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Author notes

  1. P Zhang, S Feng, G Liu and H Wang: These authors contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Luminescence and Real-Time Analytical Chemistry (Southwest University), Ministry of Education, College of Pharmaceutical Sciences, Southwest University, Chongqing, China

    P Zhang, S Feng, A Fu, H Zhu, Q Ren, X Xu & H Bai

  2. Department of Physiology, University of Alberta, Edmonton, Alberta, Canada

    P Zhang

  3. Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China

    G Liu & H Wang

  4. College of Bioengineering, Chongqing University, Chongqing, China

    B Wang

  5. Department of Biomedical Engineering, Pennsylvania State University, University Park, PA, USA

    C Dong

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Zhang, P., Feng, S., Liu, G. et al. CD82 suppresses CD44 alternative splicing-dependent melanoma metastasis by mediating U2AF2 ubiquitination and degradation. Oncogene 35, 5056–5069 (2016). https://doi.org/10.1038/onc.2016.67

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