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α-Tocopheryl succinate causes mitochondrial permeabilization by preferential formation of Bak channels

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An Erratum to this article was published on 22 October 2013

Abstract

Mitocans are drugs selectively killing cancer cells by destabilizing mitochondria and many induce apoptosis via generation of reactive oxygen species (ROS). However, the molecular events by which ROS production leads to apoptosis has not been clearly defined. In this study with the mitocan α-tocopheryl succinate (α-TOS) the role of the Bcl-2 family proteins in the mechanism of malignant cell apoptosis has been determined. Exposure of several different cancer cell lines to α-TOS increased expression of the Noxa protein, but none of the other proteins of the Bcl-2 family, an event that was independent of the cellular p53 status. α-TOS caused a profound conformational change in the pro-apoptotic protein, Bak, involving oligomerization in all cell types, and this also applied to the Bax protein, but only in non-small cell lung cancer cells. Immunoprecipitation studies indicated that α-TOS activates the two BH1-3 proteins, Bak or Bax, to form high molecular weight complexes in the mitochondria. RNAi knockdown revealed that Noxa and Bak are required for α-TOS-induced apoptosis, and the role of Bak was confirmed using Bak- and/or Bax-deficient cells. We conclude that the major events induced by α-TOS in cancer cells downstream of ROS production leading to mitochondrial apoptosis involve the Noxa-Bak axis. It is proposed that this represents a common mechanism for mitochondrial destabilization activated by a variety of mitocans that induce accumulation of ROS in the early phases of apoptosis.

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Abbreviations

BH3:

Bcl-2 homology domain-3

CII:

Complex II

Cox IV:

Cytochrome c oxidase subunit IV

DHE:

Dihydroethidium

ECL:

Enhanced chemiluminescence

MOM:

Mitochondrial outer membrane

RNAi:

RNA interference

ROS:

Reactive oxygen species

Q-PCR:

Quantitative real-time PCR

siRNA:

Short interfering RNA

STS:

Staurosporin

α-TEA:

α-Tocopheryloxyacetic acid

α-TOS:

α-Tocopheryl succinate

TRAIL:

TNF-related apoptosis-inducing ligand

VE:

Vitamin E

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Acknowledgements

Jurkat Bax−/Bak− and Jurkat Bax− cells were provided by H. Rabinowicz (University of Pittsburgh, Pittsburgh, PA, USA), H1299 and MCF7DD9 cells by Dr. B. Vojtesek (Masaryk Memorial Institute, Brno, Czech Republic), and human recombinant TRAIL by Dr. L. Andera (Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic). This work was supported in part by grants from the Australian Research Council (to J.N. and P.K.W.), the Queensland Cancer Fund, the National Breast Cancer Foundation, the Grant Agency of the Academy of Sciences of the Czech Republic KAN200520703, IAA5005220602 and IAA5005200602 to J.N, by Concept Grant AV0Z50520514 awarded by the Academy of Sciences of the Czech Republic, by a grant from Ministry of Agriculture of the Czech Republic (Grant No. MZE 0002716202) to J.T. and by a grant from the Grant agency of the Czech Republic 204/09/P632 to L.P. L.P. was supported in part by the Apoptosis Research Group (Griffith University).

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Authors and Affiliations

  1. Veterinary Research Institute, Hudcova 70, 621 00, Brno, Czech Republic

    Lubomir Prochazka & Jaroslav Turanek

  2. Apoptosis Research Group, School of Medical Science and the Griffith Institute of Health and Medical Research, Griffith University, Parklands Avenue, Southport, QLD, 4222, Australia

    Lan-Feng Dong, Ruth Freeman & Jiri Neuzil

  3. Molecular Therapy Group, Institute of Biotechnology, Czech Academy of Sciences, Prague, Czech Republic

    Karel Valis & Jiri Neuzil

  4. Genomic Research Centre, School of Medical Science, Griffith University, Southport, QLD, Australia

    Stephen J. Ralph

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  1. Lubomir Prochazka
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  2. Lan-Feng Dong
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  7. Jiri Neuzil
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Correspondence to Lubomir Prochazka or Jiri Neuzil.

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An erratum to this article is available at http://dx.doi.org/10.1007/s10495-013-0912-9.

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Prochazka, L., Dong, LF., Valis, K. et al. α-Tocopheryl succinate causes mitochondrial permeabilization by preferential formation of Bak channels. Apoptosis 15, 782–794 (2010). https://doi.org/10.1007/s10495-010-0482-z

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