Abstract
Aberrant apoptosis has been associated with the development and therapeutic resistance of cancer. Recent studies suggest that caspase deficiency/downregulation is frequently detected in different cancers. We have previously shown that caspase-3 reconstitution significantly sensitized MCF-7 cells to doxorubicin and etoposide. In contrast to the well established role of caspase-3 as an effector caspase, the focus of this study is to delineate caspase-3 induced feedback activation of the apical caspases-2, -8, -9 and -10A in doxorubicin and TNF-α induced apoptosis. Using cell-free systems we show that caspases-9 and 2 are the most sensitive, caspase-8 is less sensitive and caspase-10A is the least sensitive to caspase-3 mediated-cleavage. When apoptosis is induced by doxorubicin or TNF-α in an intact cell model, cleavage of caspases-8 and -9, but not caspase-2, was markedly enhanced by caspase-3. Caspase-3 mediated-feedback and activation of caspase-8 and -9 in MCF-7/C3 cells is further supported by an increase in the cleavage of caspase-8 and 9 substrates and cytochrome c release. These data indicate that, in addition to its function as an effector caspase, caspase-3 plays an important role in maximizing the activation of apical caspases and crosstalk between the two major apoptotic pathways. The significant impact of caspase-3 on both effector and apical caspases suggests that modulation of caspase-3 activity would be a useful approach to overcome drug resistance in clinical oncology.
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Abbreviations
- DISC:
-
death-inducing signaling complex
- CHX:
-
cycloheximide
- TnT:
-
in vitro transcription and translation
- DEVD-CHO:
-
Ac-Asp-Glu-Val-Asp-CHO
- YVAD-CHO:
-
Ac-Tyr-Val-Ala-Asp-CHO
- Z-IETD-AMC:
-
Z-Ile-Glu-Thr-Asp-AMC
- Ac-LEHD-AMC:
-
Ac-Leu-Glu-His-Asp-AMC
- PARP:
-
poly(ADP-ribose)polymerases
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XiaoHe Yang: This work was supported in part by the Career Development Award DAMD17-99-1-9180 from Department of Defense to X.H.Y.
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Yang, S., Thor, A.D., Edgerton, S. et al. Caspase-3 mediated feedback activation of apical caspases in doxorubicin and TNF-α induced apoptosis. Apoptosis 11, 1987–1997 (2006). https://doi.org/10.1007/s10495-006-0084-y
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DOI: https://doi.org/10.1007/s10495-006-0084-y