Abstract
We have previously reported that AD5-10, a novel agonistic monoclonal antibody against DR5, possessed a strong cytotoxic activity in various tumor cells, via induction of caspase-dependent and -independent signaling pathways. The present study further demonstrates that reactive oxygen species (ROS) were generated in abundance in Jurkat leukemia cells upon AD5-10 stimulation and that ROS accumulation subsequently evoked sustained activation of c-Jun N-terminal kinase (JNK), loss of mitochondrial membrane potential, and release of endonuclease G (Endo G) from mitochondria into the cytosol. The reducing agent, N-acetylcysteine (NAC), effectively inhibited the sustained activation of JNK, release of Endo G, and cell death in Jurkat cells treated by AD5–10. Moreover, a dominant-negative form of JNK (but not of p38) enhanced NF-κB activation, suppressed caspase-8 recruitment in death-inducing signaling complexes (DISCs), and reduced adverse effects on mitochondria, thereby inhibiting AD5-10-induced cell death in Jurkat leukemia cells. These data provide novel information on the DR5-mediated cell death-signaling pathway and may shed new light on effective strategies for leukemia and solid tumor therapies.
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Abbreviations
- TRAIL:
-
(tumor necrosis factor-related apoptosis-inducing ligand)
- PCD:
-
(programmed cell death)
- DD:
-
(death domain)
- TNF:
-
(tumor necrosis factor)
- FasL:
-
(Fas ligand)
- TRAIL-R:
-
(TRAIL receptor)
- DR:
-
(death receptor)
- DcR:
-
(decoy receptor)
- FADD:
-
(Fas-associated death domain protein)
- JNK:
-
(c-Jun N-terminal kinase)
- RIP:
-
(receptor-interacting protein)
- PARP:
-
(poly (ADP-ribose) polymerase)
- AIF:
-
(apoptosis-inducing factor)
- Endo G:
-
(endonuclease G)
- PBS:
-
(phosphate buffered saline)
- FCS:
-
(fetal calf serum)
- MEFs:
-
(murine embryonic fibroblasts)
- ROS:
-
(reactive oxygen species)
- NAC:
-
(N-acetylcysteine)
- GSH:
-
(glutathione)
- NAO:
-
(nonyl-acridine Orange)
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Acknowledgements
We thank Dr Shimin Hu for his generous gifts of constructs. This work was partially supported by National Natural Science Foundation of China (Grant Nos. 30571687 and 30721063) and by State Key Basic Research Program of China (Grant No. 2007CB507404).
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Chen, C., Liu, Y. & Zheng, D. An agonistic monoclonal antibody against DR5 induces ROS production, sustained JNK activation and Endo G release in Jurkat leukemia cells. Cell Res 19, 984–995 (2009). https://doi.org/10.1038/cr.2009.60
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DOI: https://doi.org/10.1038/cr.2009.60
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