Migration pattern, actin cytoskeleton organization and response to PI3K-, mTOR-, and Hsp90-inhibition of glioblastoma cells with different invasive capacities
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Simon Memmel1, Dmitri Sisario1,2, Caren Zöller1, Vanessa Fiedler1, Astrid Katzer1, Robin Heiden1, Nicholas Becker1, Lorenz Eing2, Fábio L.R. Ferreira1, Heiko Zimmermann3,4, Markus Sauer2, Michael Flentje1, Vladimir L. Sukhorukov2,* and Cholpon S. Djuzenova1,*
1Department of Radiation Oncology, University Hospital of Würzburg, Würzburg, Germany
2Department of Biotechnology and Biophysics, University of Würzburg, Würzburg, Germany
3Fraunhofer-Institut für Biomedizinische Technik, Sulzbach, Germany
4Molekulare und Zelluläre Biotechnologie/Nanotechnologie, Universität des Saarlandes, Saarbrücken, Germany
Cholpon S. Djuzenova, email: [email protected]
Vladimir L. Sukhorukov, email: [email protected]
Keywords: single-cell tracking, dSTORM, super-resolution microscopy, PTEN p53, wound healing
Received: January 12, 2017 Accepted: March 20, 2017 Published: April 05, 2017
High invasiveness and resistance to chemo- and radiotherapy of glioblastoma multiforme (GBM) make it the most lethal brain tumor. Therefore, new treatment strategies for preventing migration and invasion of GBM cells are needed. Using two different migration assays, Western blotting, conventional and super-resolution (dSTORM) fluorescence microscopy we examine the effects of the dual PI3K/mTOR-inhibitor PI-103 alone and in combination with the Hsp90 inhibitor NVP-AUY922 and/or irradiation on the migration, expression of marker proteins, focal adhesions and F-actin cytoskeleton in two GBM cell lines (DK-MG and SNB19) markedly differing in their invasive capacity. Both lines were found to be strikingly different in morphology and migration behavior. The less invasive DK-MG cells maintained a polarized morphology and migrated in a directionally persistent manner, whereas the highly invasive SNB19 cells showed a multipolar morphology and migrated randomly. Interestingly, a single dose of 2 Gy accelerated wound closure in both cell lines without affecting their migration measured by single-cell tracking. PI-103 inhibited migration of DK-MG (p53 wt, PTEN wt) but not of SNB19 (p53 mut, PTEN mut) cells probably due to aberrant reactivation of the PI3K pathway in SNB19 cells treated with PI-103. In contrast, NVP-AUY922 exerted strong anti-migratory effects in both cell lines. Inhibition of cell migration was associated with massive morphological changes and reorganization of the actin cytoskeleton. Our results showed a cell line-specific response to PI3K/mTOR inhibition in terms of GBM cell motility. We conclude that anti-migratory agents warrant further preclinical investigation as potential therapeutics for treatment of GBM.
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