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BMI-1 suppresses contact inhibition and stabilizes YAP in Ewing sarcoma

Oncogene volume 30, pages 2077–2085 (2011)Cite this article

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Abstract

The polycomb group family protein BMI-1 is overexpressed by and functions as an oncogene in many different human cancers. We have previously shown that BMI-1 promotes the tumorigenicity of Ewing sarcoma family tumors (ESFTs) and that this is mediated independently of CDKN2A repression. In this study, we have discovered that high levels of BMI-1 confer resistance to contact inhibition in ESFT cells. Using stable retroviral transduction, we evaluated the consequences of BMI-1 knockdown on the growth of CDKN2A wild-type and mutant ESFT cells in subconfluent and confluent conditions. Although knockdown of BMI-1 had no effect on proliferation in low-density cultures, at high cell densities it resulted in cell cycle arrest and death. The normal cell contact inhibition response is mediated, in large part, by the recently described Hippo pathway which functions to inhibit cell proliferation and promote cell death by inactivating the Yes-Associated Protein (YAP). Significantly, we found that YAP levels, activity and expression did not diminish in confluent ESFT cells that expressed high levels of BMI-1. In contrast, YAP expression and nuclear localization were reduced in confluent BMI-1 knockdown cells suggesting that silencing of BMI-1 restored contact inhibition by restoring normal activation of the Hippo–YAP growth-suppressor pathway. Importantly, knockdown of YAP in ESFT cells resulted in profound inhibition of cell proliferation and anchorage-independent colony formation suggesting that stabilization and continued expression of YAP is critical for ESFT growth and tumorigenicity. Together, these studies reveal a previously unrecognized link between BMI-1, contact inhibition and the Hippo–YAP pathway and suggest that resistance to contact inhibition in BMI-1 overexpressing cancer cells may be in part a result of Hippo inhibition and aberrant stabilization of YAP.

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Acknowledgements

We gratefully acknowledge Long Hung and Asim Beg for technical support, members of the Lawlor lab for helpful discussion, Dr T Triche and the Children's Oncology Cell Bank (COGCELL.org) for cell lines. This work was initiated at Childrens Hospital Los Angeles (CHLA) with support from the TJ Martell Foundation. JHH was supported by CIRM training Grant T1-00004 and ERL by NIH/NCI 1R01CA134604-01. Grant support: California Institute for Regenerative Medicine Training Grant T1-00004 (JHH), 1R01CA134604-01 (ERL).

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Authors and Affiliations

  1. Genetics, Molecular and Cellular Biology Program, University of Southern California, Los Angeles, CA, USA

    J H Hsu

  2. Division of Hematology-Oncology, Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA

    J H Hsu & E R Lawlor

  3. Department of Pathology, University of Michigan, Ann Arbor, MI, USA

    E R Lawlor

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  1. J H Hsu
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Correspondence to E R Lawlor.

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Hsu, J., Lawlor, E. BMI-1 suppresses contact inhibition and stabilizes YAP in Ewing sarcoma. Oncogene 30, 2077–2085 (2011). https://doi.org/10.1038/onc.2010.571

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