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Induction of apoptosis by enediyne antibiotic calicheamicin ϑII proceeds through a caspase-mediated mitochondrial amplification loop in an entirely Bax-dependent manner

Oncogene volume 22, pages 9107–9120 (2003)Cite this article

Abstract

Calicheamicin ϑII is a member of the enediyne class of antitumor antibiotics that bind to DNA and induce apoptosis. These compounds differ, however, from conventional anticancer drugs as they bind in a sequence-specific manner noncovalently to DNA and cause sequence-selective oxidation of deoxyriboses and bending of the DNA helix. Calicheamicin is clinically employed as immunoconjugate to antibodies directed against, for example, CD33 in the case of gemtuzumab ozogamicin. Here, we show by the use of the unconjugated drug that calicheamicin-induced apoptosis is independent from death-receptor/FADD-mediated signals. Moreover, calicheamicin triggers apoptosis in a p53-independent manner as shown by the use of p53 knockout cells. Cell death proceeds via activation of mitochondrial permeability transition, cytochrome c release and activation of caspase-9 and -3. The overexpression of Bcl-xL or Bcl-2 strongly inhibited calicheamicin-induced apoptosis. Knockout of Bax abrogated cell death after calicheamicin treatment. Thus, the activation of mitochondria and execution of cell death occur through a fully Bax-dependent mechanism. Interestingly, caspase inhibition by the pancaspase-inhibitor zVAD-fmk interfered with mitochondrial activation by calicheamicin. This places caspase activation upstream of the mitochondria and indicates that calicheamicin-triggered apoptosis is enhanced through death receptor-independent activation of the caspase cascade, that is, an amplification loop that is required for full activation of the mitochondrial pathway.

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Acknowledgements

This work was supported by grants from the Deutsche José Carreras Leukämie-Stiftung, the Verein zur Förderung der Tagesklinik and the Deutsche Krebshilfe. We thank B Vogelstein (Johns Hopkins University School of Medicine, Baltimore, MD, USA) for kindly providing HCT116 wild-type, p53 −/− and Bax −/− cells and the congeneic controls, S Fulda (University of Ulm, Germany) for kindly providing BJAB cells overexpressing Bcl-xL and K Schulze-Osthoff (University of Düsseldorf, Germany) for kindly providing Jurkat cells overexpressing Bcl-2. The excellent technical assistance of A Richter is gratefully acknowledged.

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Author notes

  1. Florian Lang & Thomas Wieder

    Present address: Department of Physiology I, Eberhard-Karls University Tubingen, Tübingen, 72076, Germany

  2. Aram Prokop and Wolf Wrasidlo: These authors contributed equally to the present study

  3. Thomas Wieder and Peter T Daniel: These authors share senior authorship

Authors and Affiliations

  1. Department of Pediatric Oncology/Hematology, University Medical Center Charité, Humboldt University of Berlin, Berlin, 13353, Germany

    Aram Prokop, Ralf Herold & Günter Henze

  2. Department of General Pediatrics, University Medical Center Charité, Humboldt University of Berlin, Berlin, 13353, Germany

    Wolf Wrasidlo & Holger Lode

  3. Department of Hematology, Oncology and Tumor Immunology, University Medical Center Charité, Humboldt University of Berlin, Berlin, 13125, Germany

    Florian Lang, Bernd Dörken, Thomas Wieder & Peter T Daniel

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Prokop, A., Wrasidlo, W., Lode, H. et al. Induction of apoptosis by enediyne antibiotic calicheamicin ϑII proceeds through a caspase-mediated mitochondrial amplification loop in an entirely Bax-dependent manner. Oncogene 22, 9107–9120 (2003). https://doi.org/10.1038/sj.onc.1207196

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