Abstract
Accumulating evidence suggests that specific isoforms of PKC may function to promote apoptosis. We show here that activation of the conventional and novel isoforms of PKC with 12-O-tetradecanoyl phorbol-13- ester (TPA) induces apoptosis in salivary acinar cells as indicated by DNA fragmentation and activation of caspase-3. TPA-induced DNA fragmentation, caspase-3 activation, and morphologic indicators of apoptosis, can be enhanced by pretreatment of cells with the calpain inhibitor, calpeptin, prior to the addition of TPA. Analysis of PKC isoform expression by immunoblot shows that TPA-induced downregulation of PKCα and PKCδ is delayed in cells pre-treated with calpeptin, and that this correlates with an increase of these isoforms in the membrane fraction of cells. TPA-induced apoptosis is accompanied by biphasic activation of the c-jun-N-terminal kinase (JNK) pathway and inactivation of the extracellular regulated kinase (ERK) pathway. Expression of constitutively activated PKCα or PKCδ, but not kinase negative mutants of these isoforms, or constitutively activated PKCε, induces apoptosis in salivary acinar cells, suggesting a role for these isoforms in TPA-induced apoptosis. These studies demonstrate that activation of PKC is sufficient for initiation of an apoptotic program in salivary acinar cells.
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Abbreviations
- β-gal:
-
β-galactosidase
- ERK:
-
extracellular regulated kinase
- JNK:
-
jun-N-terminal kinase
- PKC:
-
protein kinase C
- MAPK:
-
mitogen activated kinase
- Rb:
-
retinoblastoma protein
- TPA:
-
12-O-tetradecanoyl phorbol-13-acetate
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This research was supported by grants from the National Institutes of Health including DE12422 to MER and DK48845 to SMA.
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Reyland, M., Barzen, K., Anderson, S. et al. Activation of PKC is sufficient to induce an apoptotic program in salivary gland acinar cells. Cell Death Differ 7, 1200–1209 (2000). https://doi.org/10.1038/sj.cdd.4400744
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DOI: https://doi.org/10.1038/sj.cdd.4400744
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