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DAPK plays an important role in panobinostat-induced autophagy and commits cells to apoptosis under autophagy deficient conditions

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An Erratum to this article was published on 10 March 2016

Abstract

The histone deacetylase inhibitor (HDACi) LBH589 has been verified as an effective anticancer agent. The identification and characterization of new targets for LBH589 action would further enhance our understanding of the molecular mechanisms involved in HDACi therapy. The role of the tumor suppressor death-associated protein kinase (DAPK) in LBH589-induced cytotoxicity has not been investigated to date. Stable DAPK knockdown (shRNA) and DAPK overexpressing (DAPK+++) cell lines were generated from HCT116 wildtype colon cancer cells. LBH589 inhibited cell proliferation, reduced the long-term survival, and up-regulated and activated DAPK in colorectal cancer cells. Moreover, LBH589 significantly suppressed the growth of colon tumor xenografts and in accordance with the in vitro studies, increased DAPK levels were detected immunohistochemically. LBH589 induced a DAPK-dependent autophagy as assessed by punctuate accumulation of LC3-II, the formation of acidic vesicular organelles, and degradation of p62 protein. LBH589-induced autophagy seems to be predominantly caused by DAPK protein interactions than by its kinase activity. Caspase inhibitor zVAD increased autophagosome formation, decreased the cleavage of caspase 3 and PARP but didn’t rescue the cells from LBH589-induced cell death in crystal violet staining suggesting both caspase-dependent as well as caspase-independent apoptosis pathways. Pre-treatment with the autophagy inhibitor Bafilomycin A1 caused caspase 3-mediated apoptosis in a DAPK-dependent manner. Altogether our data suggest that DAPK induces autophagy in response to HDACi-treatment. In autophagy deficient cells, DAPK plays an essential role in committing cells to HDACi-induced apoptosis.

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Acknowledgments

The authors would like to thank Mr. Rudolf Jung and Mrs. Maria Leidenberger for excellent technical assistance and Mrs. Gabriele Krumholz for assisting in conducting animal experiments.

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Authors and Affiliations

  1. Experimental Tumorpathology, Department of Pathology, University of Erlangen-Nürnberg, Universitätsstr. 22, 91054, Erlangen, Germany

    Muktheshwar Gandesiri, Saritha Chakilam, Jelena Ivanovska, Natalya Benderska, Arndt Hartmann & Regine Schneider-Stock

  2. Institute for Surgical Research, Philipps University Marburg, Marburg, Germany

    Matthias Ocker & Pietro Di Fazio

  3. Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergology, Medical Faculty Mannheim, University of Heidelberg, Heidelberg, Germany

    Maria Feoktistova & Martin Leverkus

  4. Department of Biology, American University of Beirut, Beirut, Lebanon

    Hala Gali-Muhtasib

  5. Department of Radiation Oncology, University Hospital Goettingen, Goettingen, Germany

    Margret Rave-Fränk & Hans Christiansen

  6. Radiation Therapy and Oncology, Medical School Hanover, Hanover, Germany

    Hans Christiansen

  7. Laboratory of Molecular Imaging, Clinic of Nuclear Medicine, Erlangen, Germany

    Olaf Prante

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  1. Muktheshwar Gandesiri
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Correspondence to Regine Schneider-Stock.

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An erratum to this article is available at http://dx.doi.org/10.1007/s10495-016-1224-7.

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Gandesiri, M., Chakilam, S., Ivanovska, J. et al. DAPK plays an important role in panobinostat-induced autophagy and commits cells to apoptosis under autophagy deficient conditions. Apoptosis 17, 1300–1315 (2012). https://doi.org/10.1007/s10495-012-0757-7

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