Abstract
CD95 apoptosis resistance of tumor cells is often acquired through mutations in the death domain (DD) of one of the CD95 alleles. Furthermore, Type I cancer cells are resistant to induction of apoptosis by soluble CD95 ligand (CD95L), which does not induce efficient formation of the death-inducing signaling complex (DISC). Here, we report that tumor cells expressing a CD95 allele that lacks a functional DD, splenocytes from heterozygous lprcg mice, which express one mutated CD95 allele, and Type I tumor cells stimulated with soluble CD95L can all die through CD95 when protein synthesis or nuclear factor kappa B is inhibited. This noncanonical form of CD95-mediated apoptosis is dependent on the enzymatic activity of procaspase-8 but does not involve fully processed active caspase-8 subunits. Our data suggest that it is possible to overcome the CD95 apoptosis resistance of many tumor cells that do not efficiently form a DISC through noncanonical activation of the caspase-8 proenzyme.
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Abbreviations
- Bio-zVAD:
-
biotinylated zVAD-fmk
- CD95L:
-
CD95 ligand
- CHX:
-
cycloheximide
- DD:
-
death domain
- ΔΨm:
-
change in mitochondrial inner membrane potential
- DISC:
-
death-inducing signaling complex
- FADD:
-
Fas-associated death domain
- FLIP:
-
FLICE-like inhibitory protein
- LzCD95L:
-
leucine zipper-tagged CD95L
- lpr:
-
lymphoproliferation
- mAb:
-
monoclonal antibody
- MAPK:
-
mitogen-activated protein kinase
- NCI:
-
National Cancer Institute
- NF-κB:
-
nuclear factor kappa B
- PARP:
-
poly (ADP ribose) polymerase
- RIP:
-
receptor interacting protein
- sCD95L:
-
soluble CD95L
- TNFα:
-
tumor necrosis factor alpha
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Acknowledgements
We thank Drs P Krammer and H Walczak for providing us with anti-APO-1 and LzCD95L, respectively. We are grateful for Drs R DeMaria, A Payne and V Dixit for the Hut78 and H9 mutant cells and the crmA-expressing BJAB cells, respectively. The expression construct for human soluble CD95L was kindly provided by Dr S Nagata and the anti-mouse caspase-8 antibody by Dr A Strasser. This work was funded by the NIH Grant GM61712. A A-S was supported by the Cancer Biology Training Program 5T32CA09594 and BC. Barnhart by the DOD Breast Cancer Research Program DAMD17-03-1-0200.
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Barnhart, B., Pietras, E., Algeciras-Schimnich, A. et al. CD95 apoptosis resistance in certain cells can be overcome by noncanonical activation of caspase-8. Cell Death Differ 12, 25–37 (2005). https://doi.org/10.1038/sj.cdd.4401509
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DOI: https://doi.org/10.1038/sj.cdd.4401509
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