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Downregulation of ceramide synthase-6 during epithelial-to-mesenchymal transition reduces plasma membrane fluidity and cancer cell motility

Oncogene volume 34, pages 996–1005 (2015)Cite this article

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Abstract

Epithelial-to-mesenchymal transition (EMT) promotes cell motility, which is important for the metastasis of malignant cells, and blocks CD95-mediated apoptotic signaling triggered by immune cells and chemotherapeutic regimens. CD95L, the cognate ligand of CD95, can be cleaved by metalloproteases and released as a soluble molecule (cl-CD95L). Unlike transmembrane CD95L, cl-CD95L does not induce apoptosis but triggers cell motility. Electron paramagnetic resonance was used to show that EMT and cl-CD95L treatment both led to augmentation of plasma membrane fluidity that was instrumental in inducing cell migration. Compaction of the plasma membrane is modulated, among other factors, by the ratio of certain lipids such as sphingolipids in the membrane. An integrative analysis of gene expression in NCI tumor cell lines revealed that expression of ceramide synthase-6 (CerS6) decreased during EMT. Furthermore, pharmacological and genetic approaches established that modulation of CerS6 expression/activity in cancer cells altered the level of C16-ceramide, which in turn influenced plasma membrane fluidity and cell motility. Therefore, this study identifies CerS6 as a novel EMT-regulated gene that has a pivotal role in the regulation of cell migration.

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Acknowledgements

We are grateful to the L3 and microscopy facilities of the SFR Biosit (Université de Rennes-1, Rennes, France) for their technical assistance. We thank Nicole Therville and Julia Rochotte for their technical assistance. We also acknowledge the assistance of Dr Pauline Le Faouder and Dr Justine Bertrand-Michel from the lipidomic facilities (INSERM U1048, Toulouse, France). This work was supported by grants from the French Government managed by the French National Research Agency (ANR) and under the program ‘Investissements d’Avenir’ with reference ANR-11-LABX-0021-01-LipSTIC Labex, INCa, ARC, Cancéropole Grand Ouest, Région Bretagne, Région Bourgogne, Rennes Métropole, Ligue Contre le Cancer (Comités d’Ille-et-Vilaine/du Morbihan/des Côtes d’Armor/du Maine et Loire et les comités du Doubs/de l'Yonne), Conseil Général d'Ille-et-Vilaine, project Membratox (Grant number 32508) and the European union (FEDER).

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Author notes

  1. V Edmond, F Dufour, G Poiroux and K Shoji: These authors contributed equally to this work.

Authors and Affiliations

  1. Université de Rennes-1, Rennes, France

    V Edmond, K Shoji, M Malleter, A Fouqué, S Tauzin, O Sergent, A Penna, N Theret & P Legembre

  2. INSERM UMR 1085, IRSET, Rennes, France

    V Edmond, K Shoji, M Malleter, A Fouqué, S Tauzin, O Sergent, A Penna, N Theret & P Legembre

  3. Equipe Labellisée Ligue Contre Le Cancer, Rennes, France

    V Edmond, K Shoji, M Malleter, A Fouqué, S Tauzin, A Penna & P Legembre

  4. INSERM UMR 866, Dijon, France

    F Dufour & O Micheau

  5. INSERM UMR 1037, Toulouse, France

    G Poiroux, T Levade & B Ségui

  6. Université Toulouse III—Paul Sabatier, Toulouse, France

    G Poiroux, T Levade & B Ségui

  7. Equipe Labellisée Ligue Contre Le Cancer, Toulouse, France

    G Poiroux, T Levade & B Ségui

  8. Inserm U1052, CNRS UMR 5286, Université de Lyon, Centre de Recherche en Cancérologie de Lyon, Centre Léon Bérard Lyon, Lyon, France,

    R Rimokh

  9. INSERM U1048, Plateau Metatoul Lipidomique, Toulouse, France

    A Dupuy

  10. Centre Eugène Marquis, Rennes, France

    P Legembre

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Edmond, V., Dufour, F., Poiroux, G. et al. Downregulation of ceramide synthase-6 during epithelial-to-mesenchymal transition reduces plasma membrane fluidity and cancer cell motility. Oncogene 34, 996–1005 (2015). https://doi.org/10.1038/onc.2014.55

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