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Smac induces cytochrome c release and apoptosis independently from Bax/Bcl-xL in a strictly caspase-3-dependent manner in human carcinoma cells

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Abstract

The mitochondrial apoptosis pathway mediates cell death through the release of various pro-apoptotic factors including cytochrome c and Smac, the second mitochondrial activator of caspases, into the cytosol. Smac was shown previously to inhibit IAP proteins and to facilitate initiation of the caspase cascade upon cytochrome c release. To investigate Smac function during apoptosis and to explore Smac as an experimental cancer therapeutic, we constructed an expression system based on a single adenoviral vector containing Smac under control of the Tet-off system supplied in cis. Conditional expression of Smac induced apoptosis in human HCT116 and DU145 carcinoma cells regardless of the loss of Bax or overexpression of Bcl-xL. Nevertheless, apoptosis induced by Smac was associated with cytochrome c release and breakdown of the mitochondrial membrane potential. This indicates that Smac acts independently of Bax and Bcl-xL during initiation of apoptosis and triggers a positive feedback loop that results in Bax/Bcl-xL-independent activation of mitochondria. In caspase-proficient cells, Smac-induced apoptosis could be inhibited partially by cell-permeable LEHD (caspase-9 inhibitor) and DEVD (caspase-3 inhibitor) peptides. Furthermore, loss of caspase-3 expression in MCF-7 cells carrying a caspase-3 null mutation completely abrogated the sensitivity for Smac-induced apoptotic or nonapoptotic, necrosis-like cell death, while re-expression of caspase-3 conferred sensitivity. Altogether, caspase-3 but not caspase-9 activation was necessary for execution of Smac-induced cell death. Notably, Smac did not induce caspase-9 processing in the absence of caspase-3. Thus, caspase-9 processing occurs secondary to caspase-3 activation during Smac-induced apoptosis. Altogether, Smac is capable of circumventing defects in mitochondrial apoptosis signaling such as loss of Bax or overexpression of Bcl-xL that are frequently observed in tumor cells resistant to anticancer therapy. Consequently, Smac appears to be a promising therapeutic target in anticancer treatment.

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References

  • Anderson RD, Haskell RE, Xia H, Roessler BJ and Davidson BL . (2000). Gene Therapy, 7, 1034–1038.

  • Bosanquet AG, Sturm I, Wieder T, Essmann F, Bosanquet MI, Head DJ, Dörken B and Daniel PT . (2002). Leukemia, 16, 1035–1044.

  • Brand K, Arnold W, Bartels T, Lieber A, Kay MA, Strauss M and Dörken B . (1997). Cancer Gene Ther., 4, 9–16.

  • Chai J, Du C, Wu JW, Kyin S, Wang X and Shi Y . (2000). Nature, 406, 855–862.

  • Cosset FL, Takeuchi Y, Battini JL, Weiss RA and Collins MK . (1995). J. Virol., 69, 7430–7436.

  • Daniel PT, Sturm I, Ritschel S, Friedrich K, Dörken B, Bendzko P and Hillebrand T . (1999). Anal. Biochem., 266, 110–115.

  • Du C, Fang M, Li Y, Li L and Wang X . (2000). Cell, 102, 33–42.

  • Ekert PG, Silke J, Hawkins CJ, Verhagen AM and Vaux DL . (2001). J. Cell Biol., 152, 483–490.

  • Friedrich K, Wieder T, von Haefen C, Radetzki S, Schulze-Osthoff K, Jänicke R, Dörken B and Daniel PT . (2001). Oncogene, 20, 2749–2760.

  • Fulda S, Wick W, Weller M and Debatin KM . (2002). Nat. Med., 8, 808–815.

  • Gillissen B, Essmann F, Graupner V, Stärck L, Radetzki S, Dörken B, Schulze-Osthoff K and Daniel PT . (2003). EMBO J., 22, 3580–3590.

  • Gossen M and Bujard H . (1992). Proc. Natl. Acad. Sci. USA, 89, 5547–5551.

  • Güner D, Sturm I, Hemmati PG, Hermann S, Hauptmann S, Wurm R, Budach V, Dörken B, Lorenz M and Daniel PT . (2003). Int. J. Cancer, 103, 445–454.

  • Guo F, Nimmanapalli R, Paranawithana S, Wittman S, Griffin D, Bali P, O'Bryan E, Fumero C, Wang HG and Bhalla K . (2002). Blood, 99, 3419–3426.

  • Hemmati PG, Gillissen B, von Haefen C, Wendt J, Stärck L, Güner D, Dörken B and Daniel PT . (2002). Oncogene, 21, 3149–3161.

  • Jänicke RU, Sprengart ML, Wati MR and Porter AG . (1998). J. Biol. Chem., 273, 9357–9360.

  • Li LY, Luo X and Wang X . (2001). Nature, 412, 95–99.

  • Lorenzo HK, Susin SA, Penninger J and Kroemer G . (1999). Cell Death Differ., 6, 516–524.

  • Martinou JC and Green DR . (2001). Nat. Rev. Mol. Cell Biol., 2, 63–67.

  • Mrozek A, Petrowsky H, Sturm I, Krauss J, Hermann S, Hauptmann S, Lorenz M and Daniel P . (2003). Cell Death Differ., in press.

  • Okada H, Suh WK, Jin J, Woo M, Du C, Elia A, Duncan GS, Wakeham A, Itie A, Lowe SW, Wang X and Mak TW . (2002). Mol. Cell Biol., 22, 3509–3517.

  • Prokop A, Wieder T, Sturm I, Essmann F, Seeger K, Wuchter C, Ludwig W-D, Henze G, Dörken B and Daniel PT . (2000). Leukemia, 14, 1606–1613.

  • Radetzki S, Köhne CH, von Haefen C, Gillissen B, Sturm I, Dörken B and Daniel PT . (2002). Oncogene, 21, 227–238.

  • Raisova M, Hossini A, Eberle J, Riebeling C, Wieder T, Sturm I, Daniel PT, Orfanos CE and Geilen CC . (2001). J. Invest. Dermatol., 177, 333–340.

  • Rau B, Sturm I, Lage H, Berger S, Schneider U, Hauptmann S, Wust P, Riess H, Schlag PM, Dörken B and Daniel PT . (2003). J. Clin. Oncol., in press.

  • Roberts DL, Merrison W, MacFarlane M and Cohen GM . (2001). J. Cell Biol., 153, 221–228.

  • Schelwies K, Sturm I, Grabowski P, Scherübl H, Schindler I, Hermann S, Stein H, Buhr HJ, Riecken EO, Zeitz M, Dörken B and Daniel PT . (2002). Int. J. Cancer, 99, 589–596.

  • Slee EA, Keogh SA and Martin SJ . (2000). Cell Death Differ., 7, 556–565.

  • Sturm I, Bosanquet AG, Hermann S, Güner D, Dörken B and Daniel PT . (2003). Cell Death Differ., 10, 477–484.

  • Sturm I, Petrowsky H, Volz R, Lorenz M, Radetzki S, Hillebrand T, Wolff G, Hauptmann S, Dörken B and Daniel PT . (2001). J. Clin. Oncol., 19, 2272–2281.

  • Suzuki Y, Imai Y, Nakayama H, Takahashi K, Takio K and Takahashi R . (2001). Mol. Cell, 8, 613–621.

  • van Loo G, Saelens X, van Gurp M, MacFarlane M, Martin SJ and Vandenabeele P . (2002). Cell Death Differ., 9, 1031–1042.

  • Verhagen AM, Silke J, Ekert PG, Pakusch M, Kaufmann H, Connolly LM, Day CL, Tikoo A, Burke R, Wrobel C, Moritz RL, Simpson RJ and Vaux DL . (2002). J. Biol. Chem., 277, 445–454.

  • von Haefen C, Wieder T, Essmann F, Schulze-Osthoff K, Dörken B and Daniel PT . (2003). Oncogene, 22, 2236–2247.

  • von Haefen C, Wieder T, Gillissen B, Stärck L, Graupner V, Dörken B and Daniel PT . (2002). Oncogene, 21, 4009–4019.

  • Wu G, Chai J, Suber TL, Wu JW, Du C, Wang X and Shi Y . (2000). Nature, 408, 1008–1012.

  • Zhang L, Yu J, Park BH, Kinzler KW and Vogelstein B . (2000). Science, 290, 989–992.

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Acknowledgements

The technical assistance of Antje Richter is gratefully acknowledged. We wish to thank Dr. Bert Vogelstein, Johns Hopkins Cancer Center, Baltimore, for the kind gift of Bax-deficient HCT116 cells. This work was supported by grants from the Deutsche Forschungsgemeinschaft (DFG) (SFB 506, Da238/4-1 and Da238/4-2), and the European Union RTN network ‘Oncodeath’. AH was a recipient of a fellowship from the DFG graduate college 331. PTD, PGH and MS are supported by the Deutsche Krebshilfe.

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Authors and Affiliations

  1. Department of Hematology Oncology and Tumor Immunology, University Medical Center Charité, Campus Berlin-Buch, Humboldt University, Berlin, Germany

    Anne Hasenjäger, Bernhard Gillissen, Antje Müller, Guillaume Normand, Philipp G Hemmati, Bernd Dörken & Peter T Daniel

  2. Department of Medicine III, Johannes Gutenberg University, Mainz, Germany

    Martin Schuler

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  1. Anne Hasenjäger
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  2. Bernhard Gillissen
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  3. Antje Müller
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  4. Guillaume Normand
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  5. Philipp G Hemmati
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  7. Bernd Dörken
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  8. Peter T Daniel
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Correspondence to Peter T Daniel.

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Hasenjäger, A., Gillissen, B., Müller, A. et al. Smac induces cytochrome c release and apoptosis independently from Bax/Bcl-xL in a strictly caspase-3-dependent manner in human carcinoma cells. Oncogene 23, 4523–4535 (2004). https://doi.org/10.1038/sj.onc.1207594

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