Abstract
The HT29 adenocarcinoma is a common model of epithelial cell differentiation and colorectal cancer and its death is an oft-analyzed response to TNF family receptor signaling. The death event itself remains poorly characterized and here we have examined the involvement of caspases using pan-caspase inhibitors. zVAD-fmk did not block death of HT29 cells in response to activation of the Fas, TRAIL, TNF, TWEAK and LTβ receptors. The secondary induction of TNF or the other known bona fide death inducing ligands did not account for death following LTβ receptor activation indicating that TNF family receptors can trigger a caspase-independent death pathway regardless of the presence of canonical death domains in the receptor. To provide a frame of reference, the phenotype of HT29 death was compared to four other TNF family receptor triggered death events; Fas induced Jurkat cell apoptosis, TNF/zVAD induced L929 fibroblast necrosis, TNF induced death of WEHI 164 fibroblastoid cells and TNF/zVAD induced U937 death. The death of HT29 and U937 cells under these conditions is an intermediate form with both necrotic and apoptotic features. The efficient coupling of TNF receptors to a caspase-independent death event in an epithelial cell suggests an alternative approach to cancer therapy.
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Abbreviations
- AIF:
-
apoptosis inducing factor
- DAPI:
-
4,6 diaminophenyl-2-phenylindole
- IFNγ:
-
interferon-γ
- JNK:
-
cJun N terminal kinase
- LTβR:
-
lymphotoxin-β receptor
- MTT:
-
3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyltetrazolium bromide
- PCD:
-
programmed cell death
- PI:
-
propidium iodide
- ROS:
-
reactive oxygen species
- TNFR:
-
tumor necrosis factor receptor
- TRAF:
-
Tumor necrosis factor receptor associated factors
- TRAIL-R:
-
TRAIL receptor
- 7AAD:
-
7-amino actinomycin D
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Acknowledgements
We thank Jurg Tschopp for the Flag-TRAIL and TRAIL-R2-Ig preparations, Ya-hui Tu (Whitehead Institute) for the electron microscopy, Holly Prentice for helpful advice, Sukumari Mohan for FACS analysis, Irene Sizing for the blocking TWEAK mAb and Jennifer Gommerman, Tim Zheng and Matvey Lukashev for critical reading and comments.
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Wilson, C., Browning, J. Death of HT29 adenocarcinoma cells induced by TNF family receptor activation is caspase-independent and displays features of both apoptosis and necrosis. Cell Death Differ 9, 1321–1333 (2002). https://doi.org/10.1038/sj.cdd.4401107
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DOI: https://doi.org/10.1038/sj.cdd.4401107
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