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cAMP–PKA inhibition of SK3 channel reduced both Ca2+ entry and cancer cell migration by regulation of SK3–Orai1 complex

  • Ion channels, receptors and transporters
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Abstract

SK3 channel mediates the migration of various cancer cells. When expressed in breast cancer cells, SK3 channel forms a complex with Orai1, a voltage-independent Ca2+ channel. This SK3–Orai1 complex associates within lipid rafts where it controls a constitutive Ca2+ entry leading to cancer cell migration and bone metastases development. Since cAMP was found to modulate breast cancer cell migration, we hypothesized that this could be explained by a modulation of SK3 channel activity. Herein, we study the regulation of SK3 channel by the cAMP–PKA pathway and the consequences for SK3-dependent Ca2+ entry and cancer cell migration. We established that the beta-adrenergic receptor agonist, isoprenaline, or the direct adenylyl cyclase activator forskolin alone or in combination with the PDE4 inhibitor, CI-1044, decreased SK3 channel activity without modifying the expression of SK3 protein at the plasma membrane. Forskolin and CI-1044 reduced the SK3-dependent constitutive Ca2+ entry and the SK3-dependent migration of MDA-MB-435s cells. PKA inhibition with KT 5720 reduced: (1) the effect of forskolin and CI-1044 by 50 % on Ca2+ entry and (2) SK3 activity by inhibiting the serine phosphorylation of SK3. These cAMP-elevating agents displaced Orai1 protein outside lipid rafts in contrast to SK3, which remained in the lipid rafts fractions. All together, these results show that activation of the cAMP–PKA pathway decreases SK3 channel and SK3–Orai1 complex activities, leading to a decrease in both Ca2+ entry and cancer cell migration. This work supports the potential use of cAMP-elevating agents to reduce cancer cell migration and may provide novel opportunities to address/prevent bone metastasis.

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Acknowledgments

This work was funded by “University of Tours”, “Région Centre”, “INSERM”, “Ligue Contre le Cancer”, “Cancéropôle Grand Ouest” , Tours’ Hospital oncology association ACORT, CANCEN and ANR grant 2010 BLAN 1139-01 (to GV). Lucie Clarysse held fellowships from the “Région Centre” and “INSERM” and Maxime Gueguinou from “Région Centre”. We thanks Pr Gunther Weber and Audrey Gambade for assistance in performing lipid rafts experiments. We thank Aurore Douaud-Lecaille and Isabelle Domingo for technical assistance and Catherine Leroy for secretarial support.

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Authors and Affiliations

  1. Inserm, UMR1069 “Nutrition, Croissance et Cancer”, Tours, 37032, France

    Lucie Clarysse, Maxime Guéguinou, Marie Potier-Cartereau, Philippe Bougnoux, Stephan Chevalier, Aurélie Chantôme & Christophe Vandier

  2. Université François Rabelais, Tours, 37032, France

    Lucie Clarysse, Maxime Guéguinou, Marie Potier-Cartereau, Philippe Bougnoux, Stephan Chevalier, Aurélie Chantôme & Christophe Vandier

  3. Inserm UMR769, LabEx LERMIT, DHU TORINO, Châtenay-Malabry, 92296, France

    Grégoire Vandecasteele

  4. Faculté de Pharmacie, Université Paris Sud, Châtenay-Malabry, 92296, France

    Grégoire Vandecasteele

  5. Centre HS Kaplan, CHRU Tours, Tours, 37032, France

    Philippe Bougnoux

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  1. Lucie Clarysse
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Correspondence to Christophe Vandier.

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Lucie Clarysse, Maxime Guéguinou, Aurélie Chantôme, and Christophe Vandier contributed equally to this work.

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Clarysse, L., Guéguinou, M., Potier-Cartereau, M. et al. cAMP–PKA inhibition of SK3 channel reduced both Ca2+ entry and cancer cell migration by regulation of SK3–Orai1 complex. Pflugers Arch - Eur J Physiol 466, 1921–1932 (2014). https://doi.org/10.1007/s00424-013-1435-5

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