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Molecular and Cellular Biology, June 2001, p. 3964-3973, Vol. 21, No. 12
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.12.3964-3973.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

NF-kappa B Inducers Upregulate cFLIP, a Cycloheximide-Sensitive Inhibitor of Death Receptor Signaling

Sebastian Kreuz, Daniela Siegmund, Peter Scheurich, and Harald Wajant*

Institute of Cell Biology and Immunology, University of Stuttgart, 70569 Stuttgart, Germany

Received 10 October 2000/Returned for modification 22 November 2000/Accepted 28 March 2001

The caspase 8 homologue FLICE-inhibitory protein (cFLIP) is a potent negative regulator of death receptor-induced apoptosis. We found that cFLIP can be upregulated in some cell lines under critical involvement of the NF-kappa B pathway, but NF-kappa B activation was clearly not sufficient for cFLIP induction in all cell lines. Treatment of SV80 cells with the proteasome inhibitor N-benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG-132) or geldanamycin, a drug interfering with tumor necrosis factor (TNF)-induced NF-kappa B activation, inhibited TNF-induced upregulation of cFLIP. Overexpression of a nondegradable Ikappa Balpha mutant (Ikappa Balpha -SR) or lack of Ikappa B kinase gamma  expression completely prevented phorbol myristate acetate-induced upregulation of cFLIP mRNA in Jurkat cells. These data point to an important role for NF-kappa B in the regulation of the cFLIP gene. SV80 cells normally show resistance to TNF-related apoptosis-inducing ligand (TRAIL) and TNF, as apoptosis can be induced only in the presence of low concentrations of cycloheximide (CHX). However, overexpression of Ikappa Balpha -SR rendered SV80 cells sensitive to TRAIL-induced apoptosis in the absence of CHX, and cFLIP expression was able to reverse the proapoptotic effect of NF-kappa B inhibition. Western blot analysis further revealed that cFLIP, but not TRAF1, A20, and cIAP2, expression levels rapidly decrease upon CHX treatment. In conclusion, these data suggest a key role for cFLIP in the antiapoptotic response of NF-kappa B activation.

* Corresponding author. Mailing address: Institute of Cell Biology and Immunology, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany. Phone: 49 (711) 685 7446. Fax: 49 (711) 685 7484. E-mail: harald.wajant{at}po.uni-stuttgart.de.

Molecular and Cellular Biology, June 2001, p. 3964-3973, Vol. 21, No. 12
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.12.3964-3973.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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