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Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL- and CD95L-mediated apoptosis

Oncogene volume 27, pages 3211–3220 (2008)Cite this article

Abstract

Death ligands such as tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and certain forms of CD95L are attractive therapeutic options for metastatic melanoma. Since knowledge about the regulation of death receptor sensitivity in melanoma is sparse, we have analysed these signaling pathways in detail. The loss of CD95 or TRAIL-R1, but not of TRAIL-R2, surface expression correlated with apoptosis sensitivity in a panel of melanoma cell lines. In contrast, the expression of proteins of the apical apoptosis signaling cascade (FADD, initiator caspases-8 and cFLIP) did not predict apoptosis sensitivity. Since both TRAIL-R1 and -R2 transmit apoptotic signals, we asked whether cFLIP, highly expressed in several of the cell lines tested, is sufficient to maintain resistance to TRAIL-R2-mediated apoptosis. Downregulation of cFLIP in TRAIL-R2-positive, TRAIL-resistant IGR cells dramatically increased TRAIL sensitivity. Conversely ectopic expression of cFLIP in TRAIL-sensitive, TRAIL-R2-expressing RPM-EP melanoma cells inhibited TRAIL- and CD95L-mediated cell death. Thus, modulation of cFLIP is sufficient to sensitize TRAIL-R2-expressing cells for TRAIL. Taken together, albeit expressing all proteins necessary for death receptor-mediated apoptosis, TRAIL-R1 negative melanoma cells cannot undergo TRAIL- or CD95L-induced apoptosis due to expression of cFLIP. Hence, cFLIP represents an attractive therapeutic target for melanoma treatment, especially in combination with TRAIL receptor agonists.

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Acknowledgements

We thank R Byers, A Schwaaf and EB Bröcker for providing melanoma cell lines and H Wajant for helpful suggestions and critical reading of the manuscript. We are grateful to PH Krammer for mAbs to caspase-8 (C-15), cFLIP (NF-6) and Apo-1 (IgG1), DW Nicholson for CPP32 (Caspase-3), and X Wang for Bid antiserum. MR Sprick is supported by an EMBO Long Term fellowship (ALTF 1063-2005). We thank M Möckel for excellent technical assistance. Part of this study was funded by grants of the Wilhelm-Sander-Stiftung (2000.092.2), Deutsche Krebshilfe (106849), Exzellenzförderung Sachsen-Anhalt (N2_OGU, TP6) and DFG (Le 953/5-1) to M Leverkus. C Drewniok was supported by a NBL-3 Start-Up project (BMBF 01ZZ0407).

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  1. P Geserick and C Drewniok: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Dermatology and Venereology, Laboratory for Experimental Dermatology, Otto-von-Guericke-University Magdeburg, Magdeburg, Germany

    P Geserick, C Drewniok, M Hupe, P Diessenbacher, H Gollnick & M Leverkus

  2. Department of Apoptosis Regulation, German Cancer Research Center, Heidelberg, Germany

    T L Haas, M R Sprick & H Walczak

  3. Laboratory for Experimental Oncology and Radiation, Academisch Medisch Centrum UVA, Amsterdam, Netherlands

    M R Sprick

  4. Rudolf Virchow Center, DFG Research Center for Experimental Biomedicine University of Würzburg, Würzburg, Germany

    M P Schön

  5. Department of Dermatology, Venereology and Allergology, University of Würzburg, Würzburg, Germany

    M P Schön

  6. Medical Policlinic, University of Würzburg, Würzburg, Germany

    F Henkler

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  1. P Geserick
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Correspondence to M Leverkus.

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Geserick, P., Drewniok, C., Hupe, M. et al. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL- and CD95L-mediated apoptosis. Oncogene 27, 3211–3220 (2008). https://doi.org/10.1038/sj.onc.1210985

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