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Caspase-3 mediated feedback activation of apical caspases in doxorubicin and TNF-α induced apoptosis

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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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Authors and Affiliations

  1. Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    Shihe Yang & XiaoHe Yang

  2. Department of Pathology, University of Colorado at Denver & Health Sciences Center, Denver, CO, 80220, USA

    Ann D. Thor & Susan Edgerton

Authors
  1. Shihe Yang
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  2. Ann D. Thor
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  3. Susan Edgerton
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  4. XiaoHe Yang
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Corresponding author

Correspondence to XiaoHe Yang.

Additional information

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

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