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Histone deacetylase inhibitors induce cell death and enhance the apoptosis-inducing activity of TRAIL in Ewing’s sarcoma cells

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Abstract

Purpose

The present in vitro study was conducted to evaluate the effects of the histone deacetylase inhibitors (HDIs) suberoyl anilide hydroxamic acid (SAHA), sodium butyrate (NaB) and MS-275 applied as single agents or in combination with TRAIL in Ewing’s sarcoma.

Methods

Cytotoxic activities were assessed by cytofluorometric analysis of propidium iodide uptake, DNA fragmentation and mitochondrial depolarisation as well as by measuring caspase-9 and -3 activities. Cell-surface expression of TRAIL receptors was determined by cytofluorometry, and histone H4 acetylation was assessed by western blot.

Results

All three HDIs potently induced cell death in the two cell lines explored, SK-ES-1 and WE-68. However, they seemed to differ in their modes of action. SAHA and NaB induced mitochondrial depolarisation as well as caspase-9 and -3 activities, and their cytotoxic effects could be significantly reduced by the pan-caspase inhibitor z-VAD-fmk. MS-275 was a much weaker inducer of caspase-9 and -3 activities as well as mitochondrial injury; consistently, z-VAD-fmk had little effect on MS-275-mediated activities. Combined treatment of HDIs and TRAIL led to an additive effect in SK-ES-1 cells and a supra-additive effect in WE-68 cells. Yet, HDIs did not increase cell-surface expression of TRAIL receptor 2, but rather decreased it. Selective inhibition of caspase-8 in WE-68 cells and HDI treatment of CADO-ES-1 cells, a Ewing's sarcoma cell line deficient in caspase-8 expression, revealed that caspase-8 was not required for HDI-mediated apoptosis.

Conclusions

These results suggest that HDIs may be considered as a novel treatment strategy for Ewing’s sarcoma either applied as monotherapy or in combination with TRAIL.

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Acknowledgments

We thank J. Gänge and A. Plath for their excellent technical assistance. This work was supported by grants from the Deutsche Krebshilfe and the Alfried Krupp von Bohlen und Halbach-Stiftung.

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Authors and Affiliations

  1. Research Center of Pharmacology and Experimental Therapeutics, Ernst Moritz Arndt University, Greifswald, Germany

    Jürgen Sonnemann, Linn Dreyer & Chithra D. Palani

  2. Department of Paediatric Oncology/Haematology, Ernst Moritz Arndt University, Greifswald, Germany

    Maite Hartwig & James F. Beck

  3. Laboratory for Functional Genomics, Ernst Moritz Arndt University, Greifswald, Germany

    Le Thi Thu Hong & Uwe Völker

  4. Department of Immunology, Ernst Moritz Arndt University, Greifswald, Germany

    Ulrike Klier & Barbara Bröker

  5. Abteilung für Pädiatrische Onkologie und Hämatologie, Zentrum für Kinder- und Jugendmedizin, Soldmannstrasse 15, 17475, Greifswald, Germany

    James F. Beck

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  1. Jürgen Sonnemann
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  2. Linn Dreyer
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Correspondence to James F. Beck.

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Sonnemann, J., Dreyer, L., Hartwig, M. et al. Histone deacetylase inhibitors induce cell death and enhance the apoptosis-inducing activity of TRAIL in Ewing’s sarcoma cells. J Cancer Res Clin Oncol 133, 847–858 (2007). https://doi.org/10.1007/s00432-007-0227-8

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  • DOI: https://doi.org/10.1007/s00432-007-0227-8

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