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Ionizing radiation modulates the TRAIL death-inducing signaling complex, allowing bypass of the mitochondrial apoptosis pathway

Oncogene volume 27, pages 574–584 (2008)Cite this article

Abstract

In many tumor cell types, ionizing radiation (IR) or DNA-damaging anticancer drugs enhance sensitivity to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, which is of great clinical interest. We have investigated the molecular mechanism underlying the response to combined modality treatment in p53-mutant Jurkat T leukemic cells overexpressing Bcl-2. These cells are largely resistant to individual treatment with TRAIL or IR, but sensitive to combined treatment, in vitro as well as in vivo. We demonstrate that IR and DNA-damaging anticancer drugs enable TRAIL receptor-2 and CD95/Fas to bypass the mitochondrial pathway for effector caspase activation. This was validated by RNA interference for Bax and Bak and by overexpression of dominant-negative Caspase-9. Improved effector caspase activation was neither caused by altered expression of proapoptotic components nor by impaired activity of inhibitor of apoptosis proteins or nuclear factor-κB signaling. Rather, we found that pretreatment of cells with IR caused quantitative and qualitative changes in death receptor signaling. It strongly improved the capacity of ligand-bound receptors to recruit FADD and activate Caspase-8 and -10 in the death-inducing signaling complex, while c-FLIPL levels were unaffected.

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Abbreviations

Cyt c:

cytochrome c

DISC:

death-inducing signaling complex

dn:

dominant negative

ECL:

enhanced chemiluminescence

5-FU:

5-fluorouracil

GFP:

green fluorescent protein

IAPs:

inhibitor of apoptosis proteins

IR:

ionizing radiation

IRES:

internal ribosomal entry sequence

IZ:

isoleucine zippered

KD, knock down; L:

ligand

PI:

propidium iodide

R:

receptor

RNAi:

RNA interference

TNF:

tumor necrosis factor

TRAIL:

TNF-related apoptosis-inducing ligand

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Acknowledgements

We thank Victor Peperzak for help with microarrays and personnel of the flow cytometry and microarray facilities of The Netherlands Cancer Institute for expert technical assistance. This work was financially supported by the Dutch Cancer Society (project NKI 2004-3079).

Author information

Author notes

  1. S W G Tait

    Present address: 4Current address: Department of Immunology, St Jude Children's Research Hospital, Memphis, TN 38105-2794, USA.,

  2. E H J Wissink

    Present address: 5Current address: Division of Virology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.,

Authors and Affiliations

  1. Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    I Verbrugge, E de Vries, S W G Tait, E H J Wissink & J Borst

  2. Division of Apoptosis Regulation, German Cancer Research Center (DFKZ), Heidelberg, Germany

    H Walczak

  3. Division of Radiotherapy, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    M Verheij

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  1. I Verbrugge
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Correspondence to J Borst.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Verbrugge, I., de Vries, E., Tait, S. et al. Ionizing radiation modulates the TRAIL death-inducing signaling complex, allowing bypass of the mitochondrial apoptosis pathway. Oncogene 27, 574–584 (2008). https://doi.org/10.1038/sj.onc.1210696

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