title: Granulocyte macrophage colony-stimulating factor shows anti-apoptotic activity via the PI3K-NF-κB-HIF-1α-survivin pathway in mouse neural progenitor cells.
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.
authors.affiliation: Department of Physiology, Inha University College of Medicine, Incheon, South Korea.
authors.forename: Jung Kyoung
authors.initials: JK
authors.lastname: Choi
authors.forename: Kil Hwan
authors.initials: KH
authors.lastname: Kim
authors.forename: So Ra
authors.initials: SR
authors.lastname: Park
authors.forename: Byung Hyune
authors.initials: BH
authors.lastname: Choi
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis* / Q000235:genetics; D051034:Inhibitor of Apoptosis Proteins / Q000096:biosynthesis* / Q000235:genetics; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009025:Morpholines / Q000494:pharmacology; D016328:NF-kappa B / Q000096:biosynthesis* / Q000235:genetics; D058953:Neural Stem Cells / Q000187:drug effects / Q000378:metabolism; D058539:Phosphatidylinositol 3-Kinase / Q000096:biosynthesis* / Q000235:genetics; D000081082:Phosphoinositide-3 Kinase Inhibitors; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D000077022:Survivin
chemical_list: C117749:Birc5 protein, mouse; D002867:Chromones; D004791:Enzyme Inhibitors; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D051034:Inhibitor of Apoptosis Proteins; D009025:Morpholines; D016328:NF-kappa B; D000081082:Phosphoinositide-3 Kinase Inhibitors; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D000077022:Survivin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D058539:Phosphatidylinositol 3-Kinase
references.citation: Nature. 1989 May 4;339(6219):27-30
references.pmid: 2469962
references.citation: J Immunol. 2000 Apr 15;164(8):4286-91
references.pmid: 10754327
references.citation: Stroke. 2003 Jul;34(7):1803-8
references.pmid: 12791942
references.citation: Biochem J. 2008 Jun 15;412(3):477-84
references.pmid: 18393939
references.citation: J Cell Mol Med. 2011 Jul;15(7):1542-50
references.pmid: 20716114
references.citation: J Neurol Sci. 2009 Feb 15;277(1-2):87-97
references.pmid: 19033079
references.citation: Neurosci Lett. 2007 Jun 15;420(3):217-22
references.pmid: 17556097
references.citation: J Leukoc Biol. 1999 May;65(5):700-6
references.pmid: 10331501
references.citation: J Exp Clin Cancer Res. 2009 Feb 26;28:29
references.pmid: 19245702
references.citation: J Leukoc Biol. 2004 Nov;76(5):1047-56
references.pmid: 15328334
references.citation: Cell Signal. 1998 Oct;10(9):619-28
references.pmid: 9794243
references.citation: J Neurosurg. 2009 Jul;111(1):155-63
references.pmid: 19361262
references.citation: Biochim Biophys Acta. 2004 Mar 1;1644(2-3):149-58
references.pmid: 14996499
references.citation: J Pharmacol Exp Ther. 2002 Oct;303(1):124-31
references.pmid: 12235242
references.citation: Brain Res. 2006 May 17;1089(1):179-85
references.pmid: 16678804
references.citation: Stem Cells. 2007 Aug;25(8):2066-73
references.pmid: 17464087
references.citation: J Neurosurg Spine. 2005 Jan;2(1):55-61
references.pmid: 15658127
references.citation: Trends Neurosci. 1995 Aug;18(8):355-8
references.pmid: 7482798
references.citation: Blood. 2001 May 1;97(9):2784-90
references.pmid: 11313272
references.citation: Exp Hematol. 2010 Dec;38(12):1199-208
references.pmid: 20813154
references.citation: Apoptosis. 2011 Feb;16(2):127-34
references.pmid: 21052840
references.citation: Neurosci Lett. 2004 May 6;361(1-3):76-8
references.pmid: 15135897
references.citation: Neuroimmunomodulation. 2000;8(3):132-41
references.pmid: 11124579
references.citation: Stem Cells. 1998;16(5):301-13
references.pmid: 9766809
references.citation: Biochim Biophys Acta. 2010 Dec;1803(12):1368-75
references.pmid: 20816704
references.citation: Neuroreport. 2004 Oct 5;15(14):2161-5
references.pmid: 15371725
references.citation: Anticancer Res. 2009 Aug;29(8):2893-8
references.pmid: 19661292
references.citation: J Histochem Cytochem. 2010 Mar;58(3):265-75
references.pmid: 19901272
references.citation: J Cell Mol Med. 2003 Jul-Sep;7(3):249-57
references.pmid: 14594549
references.citation: Mol Biol Rep. 2012 Jan;39(1):185-91
references.pmid: 21567203
references.citation: J Biol Chem. 2006 Sep 8;281(36):25903-14
references.pmid: 16847054
references.citation: Tissue Eng. 2005 May-Jun;11(5-6):913-22
references.pmid: 15998231
references.citation: Clin Cancer Res. 2010 Jan 1;16(1):77-87
references.pmid: 20028744
references.citation: Neurosci Res. 2007 May;58(1):50-7
references.pmid: 17331604
references.citation: Neuroreport. 2007 Jul 16;18(11):1113-7
references.pmid: 17589309
references.citation: Eur J Neurosci. 2009 Mar;29(5):891-900
references.pmid: 19245369
references.citation: J Biol Chem. 2002 Oct 11;277(41):38205-11
references.pmid: 12149254
references.citation: Int J Oncol. 2010 Mar;36(3):593-600
references.pmid: 20126979
references.citation: Blood. 2000 Jan 15;95(2):461-9
references.pmid: 10627450
references.citation: Blood. 2004 Feb 1;103(3):820-7
references.pmid: 12920017