Abstract
ZBP-89 induces apoptosis in human gastrointestinal cancer cells through a p53-independent mechanism. To understand the apoptotic pathway regulated by ZBP-89, we identified downstream signal transduction targets. Ectopic expression of ZBP-89 induced apoptosis through the mitochondrial pathway and was accompanied by activation of all three MAP kinase subfamilies: JNK1/2, ERK1/2 and p38 MAP kinase. ZBP-89-induced apoptosis was markedly enhanced by ERK inhibition with U0126. In contrast, inhibiting JNK with a JNK1-specific peptide inhibitor or dominant-negative JNK2 expression abrogated ZBP-89-mediated apoptosis. The p38 inhibitor SB202190 had no effect on ZBP-89-induced cell death. Protein dephosphorylation assays revealed that ZBP-89 activates JNK via repression of JNK dephosphorylation. Oligonucleotide microarray analyses revealed that ectopic expression of ZBP-89 downregulated expression of the dual-specificity phosphatase MKP6. Overexpression of MKP6 blocked ZBP-89-induced JNK phosphorylation and PARP cleavage. In addition, ectopic expression of ZBP-89 repressed Bcl-xL and Mcl-1 expression, but had no effect on Bcl-2. Silencing ZBP-89 with small interfering RNA enhanced both Bcl-xL and Mcl-1 expression. Taken together, ZBP-89-mediated apoptosis occurs via a p53-independent mechanism that requires JNK activation.
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Abbreviations
- MAPK:
-
mitogen-activated protein kinase
- MEK:
-
mitogen-activated protein kinase/extracellular signal-regulated kinase kinase
- ERK:
-
extracellular signal-regulated kinase
- JNK:
-
c-Jun N-terminal kinase
- PARP:
-
poly-(ADP ribose) polymerase
- Ad:
-
adenovirus
- dnJNK2:
-
dominant-negative c-Jun N-terminal kinase 2
- p-JNK:
-
phosphorylated c-Jun N-terminal kinase
- GFP:
-
green fluorescence protein
- siRNA:
-
small interfering RNA
- RNAi:
-
RNA interference
- JIP-1:
-
JNK-interacting protein-1
- DSP:
-
dual-specificity phosphatase
- MKP:
-
MAP kinase phosphatase
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Acknowledgements
The work was supported by Public Health Service NIH Grant DK 55732 to JLM. We thank the University of Michigan Cancer Center (5P30 CA46592) for use of the flow cytometry and vector cores. We also acknowledge use of the Michigan NIDDK Biotechnology Center (U24 DK58771). We thank Dr. Bert Vogelstein (Johns Hopkins University) for providing the HCT 116 p53 wild-type and null cell lines.
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Bai, L., Yoon, S., King, P. et al. ZBP-89-induced apoptosis is p53-independent and requires JNK. Cell Death Differ 11, 663–673 (2004). https://doi.org/10.1038/sj.cdd.4401393
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DOI: https://doi.org/10.1038/sj.cdd.4401393
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