Abstract
Protein kinase-B (PKB) and its target, the forkhead transcription factor like 1 (FKHRL1)/FoxO3a, have been suggested as regulators of neurotrophin-mediated cell survival in neuronal cells. We analyzed human neuroblastoma cells and found that FKHRL1 was phosphorylated, suggesting its inactivation. To study FKHRL1 function, we infected SH-EP and NB15 cells with a 4OH-tamoxifen-regulated FKHRL1(A3)ERtm transgene. Activation of FKHRL1 promoted cytochrome-c release and caspase-dependent apoptosis. FKHRL1 induced TRAIL and the BH3-only proteins Noxa and Bim, implicating both extrinsic and intrinsic death pathways. However, expression of dnFADD did not inhibit FKHRL1-induced cell death, whereas Bcl2 protected against apoptosis. This excluded the death-receptor pathway and suggested that cell death decision is regulated by Bcl2-rheostat. Importantly, RNAi knockdown of Noxa or Bim decreased apoptosis, indicating that Noxa and Bim cooperate to mediate FKHRL1-induced cell death. We conclude that Noxa and Bim establish a connection between FKHRL1 and mitochondria, and that both BH3-only proteins are critically involved in FKHRL1-induced apoptosis in neuroblastoma.
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Abbreviations
- BDNF:
-
brain-derived neurotrophic factor
- FKHRL1:
-
forkhead transcription factor like 1
- NGF:
-
nerve growth factor
- NT-3:
-
neurotrophin-3
- NT-4:
-
neurotrophin-4
- PI3K:
-
phosphatidylinositol-3-kinase
- PKB:
-
protein kinase B
- 4OHT:
-
4OH-tamoxifen
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Acknowledgements
We thank Dr. A. Villunger for valuable discussion and for reading the manuscript, Dr. G. Nolan for the Phoenix™ packaging cell line and Dr. C. Munoz-Pinedo for donating plasmids. We also thank Cornelia Kitzbichler, Simone Wuehl and Daniela Poeham for excellent technical assistance. The work was supported by grants from the ‘Children Cancer Society of Tyrol and Vorarlberg’, the ‘Children Cancer Society South Tyrol-Regenbogen’, the ‘SVP-women-association’, the Austrian Science Fund (SFB021, part 6 and P18747 project R. Kofler), the ‘Children's Cancer Research Institute (CCRI)’ and by the OeNB Anniversary Fund (Project 11436). The Tyrolean Cancer Research Institute and this study is supported by the ‘Tiroler Landeskrankenanstalten Ges.m.b.H. (TILAK)’, the ‘Tyrolean Cancer Society’ and by the ‘Department of Health-Care, Autonomy of South Tyrol’.
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Obexer, P., Geiger, K., Ambros, P. et al. FKHRL1-mediated expression of Noxa and Bim induces apoptosis via the mitochondria in neuroblastoma cells. Cell Death Differ 14, 534–547 (2007). https://doi.org/10.1038/sj.cdd.4402017
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DOI: https://doi.org/10.1038/sj.cdd.4402017
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