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Apoptosis

, Volume 14, Issue 5, pp 711–720 | Cite as

The additional loss of Bak and not the lack of the protein tyrosine kinase p56/Lck in one JCaM1.6 subclone caused pronounced apoptosis resistance in response to stimuli of the intrinsic pathway

  • J. Rudner
  • A.-C. Mueller
  • N. Matzner
  • S. M. Huber
  • R. Handrick
  • C. Belka
  • V. Jendrossek
Original Paper

Abstract

Ionising radiation, hypoxia, and the cyclooxygenase-2 inhibitor Celecoxib are known agonists of the intrinsic apoptosis pathway that involves mitochondrial damage upstream of caspase activation. Mitochondrial integrity is regulated by the pro-apoptotic Bcl-2 protein family members Bak and Bax. Upstream of the mitochondria, many kinases and phosphatases control the apoptotic response. However, the role of the non-receptor tyrosine kinase p56/Lck during apoptosis is controversial. The present investigation demonstrate the existence of two JCaM1.6 subclones, one expressing and one deficient for Bak. The lack of p56/Lck expression in JCaM1.6 cells per se did hardly affect apoptosis induced by ionising radiation, hypoxia, or Celecoxib. Only the additional loss of Bak expression, as observed in one JCaM1.6 subclone, rendered the cells resistant. siRNA-mediated downregulation of Bak and p56/Lck mimicked the observed effects in the subclones. Earlier experiments performed with the Bak-negative clone might have lead to the wrong assumption that lack of p56/Lck alone, and not the additonal loss of Bak, was responsible for reduced sensitivity towards stimuli of the intrinsic apoptosis pathway.

Keywords

Apoptosis p56/Lck Bak Ionising radiation Celecoxib Hypoxia 

Notes

Acknowledgments

The work was supported by grants from the Federal Ministry of Education and Research (Fö. 01KS9602) and the Interdisciplinary Center of Clinical Research Tübingen (IZKF) (1468 and 1597) as well as the Deutsche Krebshilfe/Mildred-Scheel-Stiftung (107388).

Supplementary material

10495_2009_342_MOESM1_ESM.ppt (82 kb)
Supplementary material 1 (PPT 82 kb)

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Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • J. Rudner
    • 1
  • A.-C. Mueller
    • 1
  • N. Matzner
    • 2
  • S. M. Huber
    • 1
  • R. Handrick
    • 1
    • 3
  • C. Belka
    • 1
  • V. Jendrossek
    • 1
    • 3
  1. 1.Department of Radiation OncologyUniversity of TuebingenTuebingenGermany
  2. 2.Department of PhysiologyUniversity of TuebingenTuebingenGermany
  3. 3.Institute of Cell Biology, Department of Molecular Cell BiologyUniversity of Duisburg-EssenEssenGermany

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