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Loss of PTEN induces microtentacles through PI3K-independent activation of cofilin

Oncogene volume 32, pages 2200–2210 (2013)Cite this article

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Abstract

Loss of PTEN tumor suppressor enhances metastatic risk in breast cancer, although the underlying mechanisms are poorly defined. We report that homozygous deletion of PTEN in mammary epithelial cells induces tubulin-based microtentacles (McTNs) that facilitate cell reattachment and homotypic aggregation. Treatment with contractility-modulating drugs showed that McTNs in PTEN−/− cells are suppressible by controlling the actin cytoskeleton. Because outward microtubule extension is counteracted by actin cortical contraction, increased activity of actin-severing proteins could release constraints on McTN formation in PTEN−/− cells. One such actin-severing protein, cofilin, is activated in detached PTEN−/− cells that could weaken the actin cortex to promote McTNs. Expression of wild-type cofilin, an activated mutant (S3A), and an inactive mutant (S3E) demonstrated that altering cofilin phosphorylation directly affects McTNs formation. Chemical inhibition of PI3K did not reduce McTNs or inactivate cofilin in PTEN−/− cells. Additionally, knock-in expression of the two most common PI3K-activating mutations observed in human cancer patients did not increase McTNs or activate cofilin. PTEN loss and PI3K activation also caused differential activation of the cofilin regulators, LIM-kinase1 (LIMK) and Slingshot-1L (SSH). Furthermore, McTNs were suppressed and cofilin was inactivated by restoration of PTEN in the PTEN−/− cells, indicating that both the elevation of McTNs and the activation of cofilin are specific results arising from PTEN loss. These data identify a novel mechanism by which PTEN loss could remodel the cortical actin network to facilitate McTNs that promote tumor cell reattachment and aggregation. Using isogenic MCF-10A PTEN−/− and PIK3CA mutants, we have further demonstrated that there are clear differences in activation of cofilin, LIMK and SSH between PTEN loss and PI3K activation, providing a new evidence that these mutations yield distinct cytoskeletal phenotypes, which could have an impact on tumor biology.

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Acknowledgements

This work was supported by grants from the Ruth L Kirschstein-National Service Research Award, T32-HL07698 (MIV), National Cancer Institute, R01-CA124704 (SSM), Susan G Komen Foundation, KG100240 (SSM) and an Era of Hope Scholar award from the Department of Defense, BC100675 (SSM).

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Authors and Affiliations

  1. University of Maryland Marlene and Stewart Greenebaum NCI Cancer Center, Baltimore, MD, USA

    M I Vitolo, A E Boggs, R A Whipple, J R Yoon, K Thompson & S S Martin

  2. Program in Molecular Medicine, University of Maryland Graduate Program in Life Sciences, Baltimore, MD, USA

    A E Boggs, J R Yoon & S S Martin

  3. Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA, USA

    M A Matrone

  4. Department of Cell Biology, The Scripps Research Institute, La Jolla, CA, USA

    E H Cho

  5. Johns Hopkins Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA

    E M Balzer

  6. Department of Physiology, University of Maryland School of Medicine, Baltimore, MD, USA

    S S Martin

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Correspondence to S S Martin.

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The PTEN−/− cells are licensed by Horizon Discovery Ltd. (Cambridge, UK). Dr Vitolo receives compensation from the sale of these cells. The remaining authors declare no conflict of interest.

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Vitolo, M., Boggs, A., Whipple, R. et al. Loss of PTEN induces microtentacles through PI3K-independent activation of cofilin. Oncogene 32, 2200–2210 (2013). https://doi.org/10.1038/onc.2012.234

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