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Activation of the intrinsic and extrinsic pathways in high pressure-induced apoptosis of murine erythroleukemia cells
Cellular & Molecular Biology Letters volume 13, pages 49–57 (2008)
Abstract
We previously demonstrated that caspase-3, an executioner of apoptosis, is activated in the pressure-induced apoptosis of murine erythroleukemia (MEL) cells (at 100 MPa). Here, we examined the pathway of caspase-3 activation using peptide substrates and caspase inhibitors. Using the substrates of caspases-8 and -9, it was found that both are activated in cells under high pressure. The production of nuclei with sub-G1 DNA content in 100 MPa-treated MEL cells was suppressed by inhibitors of caspases-8 and -9, and pan-caspase. In 100 MPa-treated cells, pan-caspase inhibitor partially prevented the cytochrome c release from the mitochondria and the breakdown of mitochondrial membrane potential. These results suggest that the intrinsic and extrinsic pathways are activated in apoptotic signaling during the high pressure-induced death of MEL cells.
Abbreviations
- Ac-IETD-MCA:
-
acetyl-IIe-Glu-Thr-Asp-4-methylcoumaryl-7-amide
- Ac-LEHD-CHO:
-
acetyl-Leu-Glu-His-Asp-aldehyde
- Ac-LEHD-MCA:
-
acetyl-Leu-Glu-His-Asp-4-methylcoumaryl-7-amide
- CAD:
-
caspase-activated deoxyribo-nuclease
- DTT:
-
dithiothreitol
- HEPES:
-
4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid
- MEL:
-
murine erythroleukemia
- NP-40:
-
nonidet P-40
- PBS:
-
phosphate-buffered saline
- PI:
-
propidium iodide
- PMSF:
-
phenylmethanesulfonyl fluoride
- RNase A:
-
ribonuclease A
- ROS:
-
reactive oxygen species
- UV:
-
ultraviolet
- z-IETD-fmk:
-
benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone
- z-VAD-fmk:
-
benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethyl ketone
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Yamaguchi, T., Hashiguchi, K., Katsuki, S. et al. Activation of the intrinsic and extrinsic pathways in high pressure-induced apoptosis of murine erythroleukemia cells. Cell Mol Biol Lett 13, 49–57 (2008). https://doi.org/10.2478/s11658-007-0034-x
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DOI: https://doi.org/10.2478/s11658-007-0034-x