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  • Original Article
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Hierarchical involvement of Bak, VDAC1 and Bax in cisplatin-induced cell death

Oncogene volume 27, pages 4221–4232 (2008)Cite this article

Abstract

Following the screening of a battery of distinct small-interfering RNAs that target various components of the apoptotic machinery, we found that knockdown of the voltage-dependent anion channel 1 (VDAC1) was particularly efficient in preventing cell death induced by cisplatin (CDDP) in non-small cell lung cancer cells. Both the downregulation of VDAC1 and its chemical inhibition with 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid reduced the apoptosis-associated modifications induced by CDDP, including mitochondrial transmembrane potential dissipation and plasma membrane permeabilization. VDAC1 inhibition strongly reduced the CDDP-induced conformational activation of Bax, yet had no discernible effect on the activation of Bak, suggesting that VDAC1 acts downstream of Bak and upstream of Bax. Accordingly, knockdown of Bak abolished the activation of Bax, whereas Bax downregulation had no effect on Bak activation. In VDAC1-depleted cells, the failure of CDDP to activate Bax could be reversed by means of the Bcl-2/Bcl-XL antagonist ABT-737, which concomitantly restored CDDP cytotoxicity. Altogether, these results delineate a novel pathway for the induction of mitochondrial membrane permeabilization (MMP) in the course of CDDP-induced cell death that involves a hierarchical contribution of Bak, VDAC1 and Bax. Moreover, our data suggest that VDAC1 may act as a facultative regulator/effector of MMP, depending on the initial cytotoxic event.

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Abbreviations

ΔΨm:

mitochondrial transmembrane potential

BH3:

Bcl-2 homology domain 3

BRCA1:

breast cancer 1, early onset

CDDP:

cisplatin

Cyt c:

cytochrome c

DAPI:

4′,6-diamidino-2-phenylindole dihydrochloride

DIDS:

4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid

DiOC6(3):

3,3′dihexiloxalocarbocyanine iodide

ERCC1:

excision repair cross-complementing rodent repair deficiency, complementation group 1

H2O2:

hydrogen peroxide

HE:

hydroethidine

IMS:

intermembrane space

LDH:

lactate dehydrogenase

MMP:

mitochondrial membrane permeabilization

NAC:

N-acetylcysteine

NSCLC:

non-small cell lung cancer

OM:

mitochondrial outer membrane

PARP-1:

poly (ADP-ribose) polymerase 1

PI:

propidium iodide

ROS:

reactive oxygen species

siRNA:

small-interfering RNA

tBHP:

tert-butylhydroperoxide

VDAC1:

voltage-dependent anion channel 1

WST-1:

4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate.

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Acknowledgements

This work has been supported by a special grant from Ligue National contre le cancer (équipe labellisée), as well as by grants from Agence Nationale de Recherche, Agence Nationale pour la Recherche sur le Sida, Cancéropôle Ile-de-France, Fondation pour la Recherche Médicale, Institut National du Cancer, European Commission (ApoSys, RIGHT, Active p53, Trans-Death, DeathTrain, ChemoRes) and Sidaction. NT is recipient of an FRM PhD fellowship. OK is recipient of an EMBO PhD fellowship. LS is funded by a DeathTrain PhD fellowship. EM is recipient of a DeathTrain PhD student fellowship.

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Author notes

  1. N Tajeddine and L Galluzzi: These authors contributed equally to this work.

Authors and Affiliations

  1. INSERM, U848, Villejuif, France

    N Tajeddine, L Galluzzi, O Kepp, E Hangen, E Morselli, L Senovilla, N Zamzami, N Modjtahedi & G Kroemer

  2. Institut Gustave Roussy, Villejuif, France

    N Tajeddine, L Galluzzi, O Kepp, E Hangen, E Morselli, L Senovilla, N Zamzami, N Modjtahedi & G Kroemer

  3. Université Paris-Sud 11, Villejuif, France

    N Tajeddine, L Galluzzi, O Kepp, E Hangen, E Morselli, L Senovilla, N Araujo, G Pinna, N Zamzami, N Modjtahedi, A Harel-Bellan & G Kroemer

  4. CNRS FRE 2944, Laboratoire Epigénétique et Cancer – Institut André Lwoff, Villejuif, France

    N Araujo, G Pinna & A Harel-Bellan

  5. CNRS, UPR1929, Laboratoire de Biologic cellulaire et moléculaire de la secrétion – Institut de Biologie Physico-chimique, Paris, France

    N Larochette

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Correspondence to G Kroemer.

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

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Tajeddine, N., Galluzzi, L., Kepp, O. et al. Hierarchical involvement of Bak, VDAC1 and Bax in cisplatin-induced cell death. Oncogene 27, 4221–4232 (2008). https://doi.org/10.1038/onc.2008.63

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