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Identification of Key Signaling Molecules Involved in the Activation of the Swelling-Activated Chloride Current in Human Glioblastoma Cells

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Abstract

The swelling-activated chloride current (I Cl,Vol) is abundantly expressed in glioblastoma (GBM) cells, where it controls cell volume and invasive migration. The transduction pathway mediating I Cl,Vol activation in GBM cells is, however, poorly understood. By means of pharmacological and electrophysiological approaches, on GL-15 human GBM cells we found that I Cl,Vol activation by hypotonic swelling required the activity of a U73122-sensitive phospholipase C (PLC). I Cl,Vol activation could also be induced by the membrane-permeable diacylglycerol (DAG) analog OAG. In contrast, neither calcium (Ca2+) chelation by BAPTA-AM nor changes in PKC activity were able to affect I Cl,Vol activation by hypotonic swelling. We further found that R59022, an inhibitor of diacylglycerol kinase (DGK), reverted I Cl,Vol activation, suggesting the involvement of phosphatidic acid. In addition, I Cl,Vol activation required the activity of a EHT1864-sensitive Rac1 small GTPase and the resulting actin polymerization, as I Cl,Vol activation was prevented by cytochalasin B. We finally show that I Cl,Vol can be activated by the promigratory fetal calf serum in a PLC- and DGK-dependent manner. This observation is potentially relevant because blood serum can likely come in contact with glioblastoma cells in vivo as a result of the tumor-related partial breakdown of the blood–brain barrier. Given the relevance of I Cl,Vol in GBM cell volume regulation and invasiveness, the several key signaling molecules found in this study to be involved in the activation of the I Cl,Vol may represent potential therapeutic targets against this lethal cancer.

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Acknowledgments

This work was supported by grants from Fondazione Cassa di Risparmio Perugia.

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Authors and Affiliations

  1. Dipartimento di Biologia Cellulare e Ambientale, Universita’ di Perugia, Via Pascoli 1, 06123, Perugia, Italy

    Luigi Catacuzzeno, Antonio Michelucci, Luigi Sforna, Francesco Aiello, Bernard Fioretti, Emilia Castigli & Fabio Franciolini

  2. IRCCS Neuromed, Venafro, Italy

    Miriam Sciaccaluga

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  1. Luigi Catacuzzeno
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  2. Antonio Michelucci
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  3. Luigi Sforna
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  4. Francesco Aiello
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Correspondence to Luigi Catacuzzeno.

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Luigi Catacuzzeno and Antonio Michelucci have equally contributed to this work.

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Catacuzzeno, L., Michelucci, A., Sforna, L. et al. Identification of Key Signaling Molecules Involved in the Activation of the Swelling-Activated Chloride Current in Human Glioblastoma Cells. J Membrane Biol 247, 45–55 (2014). https://doi.org/10.1007/s00232-013-9609-9

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