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  • Original Article
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7-Bromoindirubin-3′-oxime induces caspase-independent cell death

Oncogene volume 25, pages 6304–6318 (2006)Cite this article

Abstract

Indirubin, an isomer of indigo, is a reported inhibitor of cyclin-dependent kinases (CDKs) and glycogen synthase kinase-3 (GSK-3) as well as an agonist of the aryl hydrocarbon receptor (AhR). Indirubin is the active ingredient of a traditional Chinese medicinal recipe used against chronic myelocytic leukemia. Numerous indirubin analogs have been synthesized to optimize this promising kinase inhibitor scaffold. We report here on the cellular effects of 7-bromoindirubin-3′-oxime (7BIO). In contrast to its 5-bromo- and 6-bromo- isomers, and to indirubin-3′-oxime, 7BIO has only a marginal inhibitory activity towards CDKs and GSK-3. Unexpectedly, 7BIO triggers a rapid cell death process distinct from apoptosis. 7-Bromoindirubin-3′-oxime induces the appearance of large pycnotic nuclei, without classical features of apoptosis such as chromatin condensation and nuclear fragmentation. 7-Bromoindirubin-3′-oxime-induced cell death is not accompanied by cytochrome c release neither by any measurable effector caspase activation. Furthermore, the death process is not altered either by the presence of Q-VD-OPh, a broad-spectrum caspase inhibitor, or the overexpression of Bcl-2 and Bcl-XL proteins. Neither AhR nor p53 is required during 7BIO-induced cell death. Thus, in contrast to previously described indirubins, 7BIO triggers the activation of non-apoptotic cell death, possibly through necroptosis or autophagy. Although their molecular targets remain to be identified, 7-substituted indirubins may constitute a new class of potential antitumor compounds that would retain their activity in cells refractory to apoptosis.

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Abbreviations

AhR:

aryl hydrocarbon receptor

ARNT:

aryl hydrocarbon receptor nuclear translocator

BIO:

bromoindirubin-3′-oxime

CDK:

cyclin-dependent kinase

FCS:

fetal calf serum

FLT-3:

FMS-like tyrosine kinase 3

GSK-3:

glycogen synthase kinase-3

IFNα:

interferon α

IO:

indirubin-3′-oxime

LDH:

lactate dehydrogenase

MeBIO:

1-methyl-bromoindirubin-3′-oxime

MeIO:

1-methyl-indirubin-3′-oxime

MTS:

3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium

RA:

retinoic acid

ROS:

reactive oxygen species

STS:

staurosporine

TCDD:

2,3,7,8-tetrachlorodibenzo-p-dioxin

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Acknowledgements

We thank our colleagues for providing reagents: Dr Martin Goëttlicher, Dr Steve Safe, Dr Bert Vogelstein. This research was supported a grant from the EEC (FP6-2002-Life Sciences & Health, PRO-KINASE Research Project) (LM), and a ‘Cancéropole Grand-Ouest’ grant (LM). KB was supported by a fellowship from the ‘Ministère de la Recherche’. The Molecular Pharmacology Group thank the ‘Instituto de Salud Carlos III’ (PI041488, 2005-2007) for financial support.

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Author notes

  1. J Ribas and K Bettayeb: These two authors contributed equally to this work.

Authors and Affiliations

  1. C.N.R.S., Cell Cycle Group and UPS2682, Station Biologique, Bretagne, France

    J Ribas, K Bettayeb, Y Ferandin, M Knockaert & L Meijer

  2. Molecular Pharmacology Laboratory, DCMB, School of Medicine, University of Lleida, Lleida, Catalunya, Spain

    J Ribas, X Garrofé-Ochoa & J Boix

  3. ProQinase GmbH, Freiburg, Germany

    F Totzke & C Schächtele

  4. INSERM U 482, 184, rue du Faubourg Saint Antoine, Paris, Cedex, France

    J Mester

  5. Department of Pharmacy, Division of Pharmacognosy and Natural Products Chemistry, University of Athens, Panepistimiopolis Zografou, Athens, Greece

    P Polychronopoulos, P Magiatis & A-L Skaltsounis

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Correspondence to L Meijer.

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Ribas, J., Bettayeb, K., Ferandin, Y. et al. 7-Bromoindirubin-3′-oxime induces caspase-independent cell death. Oncogene 25, 6304–6318 (2006). https://doi.org/10.1038/sj.onc.1209648

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