Abstract
We investigated the role of some key regulators of cell cycle in the activation of caspases during apoptosis of insulin-secreting cells after sustained depletion of GTP by a specific inosine 5′-monophosphate dehydrogenase inhibitor, mycophenolic acid (MPA). p21Waf1/Cip1 was significantly increased following MPA treatment, an event closely correlated with the time course of caspase activation under the same conditions. MPA-induced p21Waf1/Cip1 was not mediated by p53, since p53 mass was gradually reduced over time of MPA treatment. The increment of p21Waf1/Cip1 by MPA was further enhanced in the presence of a pan-caspase inhibitor, indicating that the increased p21Waf1/Cip1 may occur prior to caspase activation. This notion of association of p21Waf1/Cip1 accumulation with caspase activation and apoptosis was substantiated by using mimosine, a selective p21Waf1/Cip1 inducer independent of p53. Mimosine, like MPA, also increased p21Waf1/Cip1, promoted apoptosis and simultaneously increased the activity of caspases. Furthermore, knocking down of p21Waf1/Cip1 transfection of siRNA duplex inhibited caspase activation and apoptosis due to GTP depletion. In contrast to p21Waf1/Cip1, a reduction in p27Kip1 occurred in MPA-treated cells. These results indicate that p21Waf1/Cip1 may act as an upstream signal to block mitogenesis and activate caspases which in turn contribute to induction of apoptosis.
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Abbreviations
- CDK:
-
cyclin-dependent kinase
- CKI:
-
cyclin-dependent kinase inhibitor
- GN:
-
guanine nucleotide
- IMPDH:
-
inosine 5′-monophosphate dehydrogenase
- MPA:
-
mycophenolic acid
- DTT:
-
dithiothreitol
- CHAPS:
-
3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
- PI:
-
propidium iodide
- PMSF:
-
phenylmethylsulfonyl fluoride
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Acknowledgements
We are grateful to Dr. RP Robertson for providing HIT-T15 cells. This work was supported by the National Medical Research Council of Singapore (R-364-000-004-213 and NMRC/0540/2001 to GDL). JH was a recipient of Research Scholarship from the National University of Singapore.
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Edited by V De Laurenzi
Portions of this work were presented in the 61st (in Philadelphia, June 2000) Annual Scientific Meetings of the American Diabetes Association.
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Huo, J., Metz, S. & Li, G. p53-independent induction of p21waf1/cip1 contributes to the activation of caspases in GTP-depletion-induced apoptosis of insulin-secreting cells. Cell Death Differ 11, 99–109 (2004). https://doi.org/10.1038/sj.cdd.4401322
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DOI: https://doi.org/10.1038/sj.cdd.4401322
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