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MycN sensitizes neuroblastoma cells for drug-induced apoptosis

Oncogene volume 18, pages 1479–1486 (1999)Cite this article

Abstract

Amplification of the MYCN gene is found in a large proportion of neuroblastoma and considered as an adverse prognostic factor. To investigate the effect of ectopic MycN expression on the susceptibility of neuroblastoma cells to cytotoxic drugs we used a human neuroblastoma cell line harboring tetracycline-controlled expression of MycN. Neither conditional expression of MycN alone nor low drug concentrations triggered apoptosis. However, when acting in concert, MycN and cytotoxic drugs efficiently induced cell death. Apoptosis depended on mitochondrial permeability transition and activation of caspases, since the mitochondrion-specific inhibitor bongkrekic acid and the caspase inhibitor ZVAD-fmk almost completely abrogated apoptosis. Loss of mitochondrial transmembrane potential and release of cytochrome c from mitochondria preceded activation of caspase-8 and caspase-3 and cleavage of PARP. CD95 expression was upregulated by treatment with cytotoxic drugs, while MycN cooperated with cytotoxic drugs to increase sensitivity to CD95-induced apoptosis and enhancing CD95-L expression. MycN overexpression and cytotoxic drugs also synergized to induce p53 and Bax protein expression, while Bcl-2 and Bcl-XL protein levels remained unchanged. Since amplification of MYCN is usually associated with a poor prognosis, these findings suggest that dysfunctions in apoptosis pathways may be a mechanism by which MycN-induced apoptosis of neuroblastoma cells is inhibited.

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Abbreviations

BA:

bongkrekic acid

CD95-L:

CD95 ligand

DiOC6(3):

3,3′-dihexyloxacarbocyanide iodide

Doxo:

Doxorubicin

ECL:

enhanced chemiluminescence

FACS:

fluorescence-activated cell-sorting

ICE:

interleukin 1β-converting enzyme

ΔΨm:

mitochondrial transmembrane potential

PARP:

poly(ADP-ribose) polymerase

Z-VAD.fmk:

benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone

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Acknowledgements

This work has been partially supported by grants from the Deutsche Forschungsgemeinschaft, the Bundesministerium für Forschung and Technologie, Bonn, the Tumor Center Heidelberg/Mannheim, The Deutsche Leukämieforschungshilfe (to K-MD), the Dr Mildred Scheel Stiftung and the Fördergemeinschaft Kinderkrebs-Neuroblastom-Forschung e.V. (to MS).

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Authors and Affiliations

  1. University Children's Hospital, Prittwitzstr. 43, D-89075, Ulm, Germany

    Simone Fulda & Klaus-Michael Debatin

  2. Department of Cytogenetics, German Cancer Research Center, Im Neuenheimer Feld 280, Heidelberg, D-69120, Germany

    Werner Lutz & Manfred Schwab

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  1. Simone Fulda
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  2. Werner Lutz
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  4. Klaus-Michael Debatin
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Fulda, S., Lutz, W., Schwab, M. et al. MycN sensitizes neuroblastoma cells for drug-induced apoptosis. Oncogene 18, 1479–1486 (1999). https://doi.org/10.1038/sj.onc.1202435

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