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Activation of the intrinsic and extrinsic pathways in high pressure-induced apoptosis of murine erythroleukemia cells


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.









caspase-activated deoxyribo-nuclease




4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid


murine erythroleukemia


nonidet P-40


phosphate-buffered saline


propidium iodide


phenylmethanesulfonyl fluoride

RNase A:

ribonuclease A


reactive oxygen species




benzyloxycarbonyl-Ile-Glu(OMe)-Thr-Asp(OMe)-fluoromethyl ketone


benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethyl ketone


  1. Okada, H. and Mak, T.W. Pathways of apoptotic and non-apoptotic death in tumour cells. Nature Rev. Cancer 4 (2004) 592–603.

    Article  CAS  Google Scholar 

  2. Berghe, T.V., van Loo, G., Saelens, X., van Gurp, M., Brouckaert, G., Kalai, M., Declercq, W. and Vandenabeele, P. Differential signaling to apoptotic and necrotic cell death by Fas-associated death domain protein FADD. J. Biol. Chem. 279 (2004) 7925–7933.

    Article  CAS  Google Scholar 

  3. Majno, G. and Joris, I. Apoptosis, oncosis, and necrosis: an overview of cell death. Am. J. Pathol. 146 (1995) 3–15.

    PubMed  CAS  Google Scholar 

  4. Green, D.R. Apoptotic pathways: paper wraps stone blunts scissors. Cell 102 (2000) 1–4.

    Article  PubMed  CAS  Google Scholar 

  5. Scoltock, A.B. and Cidlowski, J.A. Activation of intrinsic and extrinsic pathways in apoptotic signaling during UV-C-induced death of Jurkat cells: the role of caspase inhibition. Exp. Cell Res. 297 (2004) 212–223.

    Article  PubMed  CAS  Google Scholar 

  6. Rudner, J., Lepple-Wienhues, A., Budach, W., Berschauer, J., Friedrich, B., Wesselborg, S., Schulze-Osthoff, K. and Belka, C. Wild-type, mitochondrial and ER-restricted Bcl-2 inhibit DNA damage-induced apoptosis but do not affect death receptor-induced apoptosis. J. Cell Sci. 114 (2001) 4164–4172.

    Google Scholar 

  7. Matsuzawa, A. and Ichijo, H. Molecular mechanisms of the decision between life and death: regulation of apoptosis by apoptosis signal-regulating kinase 1. J. Biochem. 130 (2001) 1–8.

    PubMed  CAS  Google Scholar 

  8. Schuler, M., Bossy-Wetzel, E., Goldstein, J.C., Fitzgerald, P. and Green, D.R. p53 induces apoptosis by caspase activation through mitochondrial cytochrome c release. J. Biol. Chem. 275 (2000) 7337–7342

    Article  PubMed  CAS  Google Scholar 

  9. Rosse, T., Olivier, R., Monney, L., Rager, M., Conus, S., Fellay, I., Jansen, B. and Borner, C. Bcl-2 prolongs cell survival after Bax-induced release of cytochrome c. Nature 391 (1998) 496–499.

    Article  PubMed  CAS  Google Scholar 

  10. Adams, J. M. and Cory, S. Life-or-death decisions by the Bcl-2 protein family. Trends Biochem. Sci. 26 (2001) 61–66.

    Article  PubMed  CAS  Google Scholar 

  11. Take, J., Yamaguchi, T., Mine, N. and Terada, S. Caspase activation in high-pressure-induced apoptosis of murine erythroleukemia cells. Jpn. J. Physiol. 51 (2001) 193–199.

    Article  PubMed  CAS  Google Scholar 

  12. Takano, K.J., Takano, T., Yamanouchi, Y. and Satou, T. Pressure-induced apoptosis in human lymphoblasts. Exp. Cell Res. 235 (1997)155–160.

    Article  PubMed  CAS  Google Scholar 

  13. Matsumoto, M., Yamaguchi, T., Fukumaki, Y., Yasunaga, R. and Terada, S. High pressure induces G2 arrest in murine erythroleukemia cells. J. Biochem. 123 (1998) 87–93.

    PubMed  CAS  Google Scholar 

  14. Luo, X., Budihardjo, I., Zou, H., Slaughter, C. and Wang, X. Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94 (1998) 481–490.

    Article  PubMed  CAS  Google Scholar 

  15. Sohn, D., Schulze-Osthoff, K. and Janicke, R.U. Caspase-8 can be activated by interchain proteolysis without receptor-triggered dimerization during drug-induced apoptosis. J. Biol. Chem. 280 (2005) 5267–5273.

    Article  PubMed  CAS  Google Scholar 

  16. Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A. and Nagata, S. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391 (1998) 43–50.

    Article  PubMed  CAS  Google Scholar 

  17. Hassepass, I., Voit, R. and Hoffmann, I. Phosphorylation at serine 75 is required for UV-mediated degradation of human Cdc 25A phosphatase at the S-phase checkpoint. J. Biol. Chem. 278 (2003) 29824–29829.

    Article  PubMed  CAS  Google Scholar 

  18. Lee, J-H. and Paull, T.T. Direct activation of the ATM protein kinase by the Mre11/Rad 50/Nbs1 complex. Science 304 (2004) 93–96.

    Article  PubMed  CAS  Google Scholar 

  19. Ward, I. M., Minn, K. and Chen J. UV-induced ataxia-telangiectasia-mutated and Rad3-related (ATR) activation requires replication stress. J. Biol. Chem. 279 (2004) 9677–9680.

    Article  PubMed  CAS  Google Scholar 

  20. Takahashi, H., Yamaguchi, T., Koga, M., Kageura, H. and Terada, S. DNA replication reaction in Xenopus cell-free system is suppressed by high pressure. Cell. Mol. Biol. Lett. 9 (2004) 423–427.

    PubMed  CAS  Google Scholar 

  21. Weber, G. and Drickamer, H.G. The effect of high pressure upon proteins and other biomolecules. Q. Rev. Biophys. 16 (1983) 89–112.

    Article  PubMed  CAS  Google Scholar 

  22. Aertsen, A., DeSpiegeleer, P, Vanoirbeek, K., Lavilla, M. and Michiels, C.W. Induction of oxidative stress by high hydrostatic pressure in Escherichia coli. Appl. Environ. Microbiol. 71 (2005) 2226–2231.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Takeo Yamaguchi.

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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).

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