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Loss of giant obscurins from breast epithelium promotes epithelial-to-mesenchymal transition, tumorigenicity and metastasis

Oncogene volume 34, pages 4248–4259 (2015)Cite this article

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Abstract

Obscurins, encoded by the single OBSCN gene, are giant cytoskeletal proteins with structural and regulatory roles. The OBSCN gene is highly mutated in different types of cancers. Loss of giant obscurins from breast epithelial cells confers them with a survival and growth advantage, following exposure to DNA-damaging agents. Here we demonstrate that the expression levels and subcellular distribution of giant obscurins are altered in human breast cancer biopsies compared with matched normal samples. Stable clones of non-tumorigenic MCF10A cells lacking giant obscurins fail to form adhesion junctions, undergo epithelial-to-mesenchymal transition and generate >100-μm mammospheres bearing markers of cancer-initiating cells. Obscurin-knockdown MCF10A cells display markedly increased motility as a sheet in 2-dimensional (2D) substrata and individually in confined spaces and invasion in 3D matrices. In line with these observations, actin filaments redistribute to extending filopodia where they exhibit increased dynamics. MCF10A cells that stably express the K-Ras oncogene and obscurin short hairpin RNA (shRNA), but not scramble control shRNA, exhibit increased primary tumor formation and lung colonization after subcutaneous and tail vein injections, respectively. Collectively, our findings reveal that loss of giant obscurins from breast epithelium results in disruption of the cell–cell contacts and acquisition of a mesenchymal phenotype that leads to enhanced tumorigenesis, migration and invasiveness in vitro and in vivo.

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Acknowledgements

This work was supported by a Pilot Grant from The Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins Institute for NanoBioTechnology (U54CA143868 to AK-K) and by awards from the National Science Foundation (Award NSF-1159823 to KK), the National Cancer Institute (Awards U54-CA143868 to KK; R01-CA186286 to KK; T32-CA130840 to KMS; F32-CA177756 to KMS; R01-CA154624 to SM and K01-CA166576 to MIV) and the Kleberg Foundation (to KK). Portions of this work are included in a United States patent pending, 14/221,755, filed on 21 March 2014.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, USA

    M Shriver & A Kontrogianni-Konstantopoulos

  2. Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA

    K M Stroka & K Konstantopoulos

  3. Johns Hopkins Institute for NanoBioTechnology, The Johns Hopkins University, Baltimore, MD, USA

    K M Stroka & K Konstantopoulos

  4. Johns Hopkins Physical Sciences-Oncology Center, The Johns Hopkins University, Baltimore, MD, USA

    K M Stroka & K Konstantopoulos

  5. Marlene and Stewart Greenebaum National Cancer Institute Cancer Center, University of Maryland School of Medicine, Baltimore, MD, USA

    M I Vitolo, S Martin & A Kontrogianni-Konstantopoulos

  6. Department of Molecular and Comparative Pathobiology, The Johns Hopkins University and The Sidney Kimmel Comprehensive Cancer Center, Baltimore, MD, USA

    D L Huso

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Correspondence to A Kontrogianni-Konstantopoulos.

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Shriver, M., Stroka, K., Vitolo, M. et al. Loss of giant obscurins from breast epithelium promotes epithelial-to-mesenchymal transition, tumorigenicity and metastasis. Oncogene 34, 4248–4259 (2015). https://doi.org/10.1038/onc.2014.358

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