Abstract
Most human malignant glioma cell lines are susceptible to CD95 ligand (CD95L)-induced apoptosis. Here, we report that glioma cells are also susceptible to the cytotoxic effects of exogenous C2-ceramide. This form of cell death exhibits some morphological features of apoptosis as assessed by electron microscopy, but is unaffected by the broad spectrum caspase inhibitor, zVAD-fmk. Further, CD95L-induced apoptosis is synergistically enhanced by coexposure of the glioma cells to CD95L and C2-ceramide. CD95L-induced caspase 3-like activity, cytochrome c release and cleavage of caspases 3, 8, 9 and poly(ADP-ribose)polymerase (PARP) increase substantially after cotreatment with CD95L and C2-ceramide compared with CD95L treatment alone. None of these events occur in response to cytotoxic concentrations of C2-ceramide alone. C2-ceramide does not alter CD95 expression. Gene transfer-mediated enhancement of CD95 expression results not only in increased susceptibility to CD95L, but also in increased sensitivity to C2-ceramide. We conclude that (i) synergistic induction of apoptosis by C2-ceramide and CD95L depend on a cross-talk between the two signal transduction pathways and that (ii) C2-ceramide, independently of its sensitizing effects on CD95-dependent caspase activation, is also capable of triggering an apoptotic signaling cascade that is unaffected by zVAD-fmk-mediated caspase inhibition, but promoted by high levels of CD95 expression. Cell Death and Differentiation (2001) 8, 595–602
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Abbreviations
- Ac-DEVD-amc:
-
acetyl-DEVD-aminomethyl coumarin
- zVAD-FMK:
-
benzoyl-VAD-fluoromethylketone
- TCA:
-
trichloracetic acid
- CD95L:
-
CD95 ligand
- PBS:
-
phosphate-buffered saline
- DAPI:
-
4′6-diamidine-2′-phenylindole dihydrochloride
- TNF:
-
tumor necrosis factor
- DISC:
-
death inducing signaling complex
- RIP:
-
receptor-interacting protein
- AIF:
-
apoptosis-inducing factor
- JNK:
-
Jun-N-terminal kinase
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Acknowledgements
We thank Y Lazebnik (Cold Spring Harbor, NY, USA) for caspase 9 antibody and PH Krammer (Heidelberg, Germany) for caspase 8 antibody. This study was supported by the German Cancer Council (Deutsche Krebshilfe) (10-1287-We 2).
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Wagenknecht, B., Roth, W., Gulbins, E. et al. C2-ceramide signaling in glioma cells: synergistic enhancement of CD95-mediated, caspase-dependent apoptosis. Cell Death Differ 8, 595–602 (2001). https://doi.org/10.1038/sj.cdd.4400848
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DOI: https://doi.org/10.1038/sj.cdd.4400848
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