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Granulocyte Macrophage Colony-Stimulating Factor Shows Anti-apoptotic Activity via the PI3K–NF-κB–HIF-1α–Survivin Pathway in Mouse Neural Progenitor Cells

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Abstract

Granulocyte macrophage-colony stimulating factor (GM-CSF) is a hematopoietic cytokine that plays a crucial role in regulating the proliferation, differentiation, and survival of hematopoietic cells. Recent studies have shown that GM-CSF also has anti-apoptotic effects and regulates the expression of anti-apoptotic genes including Bcl-2 family proteins in neuronal cells in vitro and in vivo. However, the mechanism underlying the anti-apoptotic function of GM-CSF is not well understood. In the present work, we examined the role of phosphoinositide 3-kinase (PI3K)–AKT signal pathway in the anti-apoptotic activity of GM-CSF in mouse neural progenitor cells (NPCs). In terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, the anti-apoptotic effect of GM-CSF (apoptotic population of approximately 8.17 %) on staurosporine-induced apoptosis of NPCs (31.09 %) was significantly blocked by LY294002, an inhibitor of PI3K signal (24.04 %). We found that the PI3K–AKT signal pathway induced by GM-CSF treatment activated nuclear factor κB (NF-κB) and increased the expression of hypoxia-inducible factor 1α (HIF-1α) in normoxic conditions. Analyses using specific small interfering RNAs (siRNAs) showed that NF-κB was an upstream molecule of HIF-1α and activated its expression at the mRNA level. Further analyses using the siRNAs and chromatin immunoprecipitation (ChIP) showed that HIF-1α was responsible for the induced expression of survivin, a member of the inhibitor of apoptosis proteins (IAPs). Each of the specific siRNAs for NF-κB, HIF-1α, and survivin inhibited significantly the anti-apoptotic activity of GM-CSF on the staurosporine-induced apoptosis in NPCs in TUNEL assays. The results of this study showed the downstream signals and mechanism of PI3K/AKT-mediated anti-apoptotic activity of GM-CSF in NPCs, particularly revealing the role of the NF-κB–HIF-1α–survivin cascade.

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Acknowledgments

This study was supported by a grant from the Ministry of Education, Science & Technology (2009–0079196) and the Inha University Research Grant, Republic of Korea.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Physiology, Inha University College of Medicine, Incheon, South Korea

    Jung Kyoung Choi, Kil Hwan Kim & So Ra Park

  2. Division of Biomedical and Bioengineering Sciences, Inha University College of Medicine, Jungseok B/D, 7-206, Sinheung-dong 3-ga, , Jung-gu,, Incheon,, 400-712, South Korea

    Byung Hyune Choi

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  1. Jung Kyoung Choi
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Correspondence to Byung Hyune Choi.

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Jung Kyoung Choi and Kil Hwan Kim contributed equally to this work.

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Choi, J.K., Kim, K.H., Park, S.R. et al. Granulocyte Macrophage Colony-Stimulating Factor Shows Anti-apoptotic Activity via the PI3K–NF-κB–HIF-1α–Survivin Pathway in Mouse Neural Progenitor Cells. Mol Neurobiol 49, 724–733 (2014). https://doi.org/10.1007/s12035-013-8550-3

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