Notch signaling represses hypoxia-inducible factor-1α-induced activation of Wnt/β-catenin signaling in osteoblasts under cobalt-mimicked hypoxia

Li,Chen‑Tian; Liu,Jian‑Xiu; Yu,Bo; Liu,Rui; Dong,Chao; Li,Song‑Jian

doi:10.3892/mmr.2016.5324

Notch signaling represses hypoxia-inducible factor-1α-induced activation of Wnt/β-catenin signaling in osteoblasts under cobalt-mimicked hypoxia

Authors:
- Chen‑Tian Li
- Jian‑Xiu Liu
- Bo Yu
- Rui Liu
- Chao Dong
- Song‑Jian Li
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Affiliations: Department of Orthopedics, Zhujiang Hospital of Southern Medical University, Guangzhou, Guangdong 510282, P.R. China
Published online on: May 23, 2016 https://doi.org/10.3892/mmr.2016.5324
Pages: 689-696
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The modification of Wnt and Notch signaling pathways by hypoxia, and its association with osteoblast proliferation and apoptosis remain to be fully elucidated. To investigate Wnt-Notch crosstalk, and its role in hypoxia-induced osteoblast proliferation and apoptosis regulation, the present study investigated the effects of cobalt‑mimicked hypoxia on the mouse pre-osteoblast-like cell line, MC3T3‑E1, when the Notch signals were repressed using a γ‑secretase inhibitor DAPT. The data showed that the cobalt‑mimicked hypoxia suppressed cell proliferation under normal conditions, but increased cell proliferation under conditions of Notch repression, in a concentration‑dependent manner. The results of western blot and reverse transcription‑quantitative polymerase chain reaction analyses showed that the cobalt treatment increased the levels of activated β‑catenin protein and the expression levels of the target genes, axis inhibition protein 2 and myelocytomatosis oncogene, under DAPT‑induced Notch repression. However, no significant changes were found in the expression levels of the Notch intracellular domain protein or the Notch target gene, hes1. In a β‑catenin gene‑knockdown experiment, the proliferation of the MC3T3‑E1 cells under hypoxia were decreased by DAPT treatment, and knockdown of the expression of hypoxia‑inducible factor‑1α (HIF‑1α) suppressed the cobalt‑induced increase in Wnt target gene levels. No significant difference in cell proliferation rate was found following DAPT treatment when the expression of HIF‑1α was knocked down. The results of the present study showed the opposing effects of Wnt and Notch signaling under cobalt‑mimicked hypoxia, which were partially regulated by HIF‑1α, The results also showed that osteoblast proliferation was dependent on Wnt-Notch signal crosstalk.

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1	Collu GM, Hidalgo-Sastre A and Brennan K: Wnt-Notch signalling crosstalk in development and disease. Cell Mol Life Sci. 71:3553–3567. 2014. View Article : Google Scholar : PubMed/NCBI
2	Hayward P, Kalmar T and Arias AM: Wnt/Notch signalling and information processing during development. Development. 135:411–424. 2008. View Article : Google Scholar : PubMed/NCBI
3	Deregowski V, Gazzerro E, Priest L, Rydziel S and Canalis E: Notch 1 overexpression inhibits osteoblastogenesis by suppressing Wnt/beta-catenin but not bone morphogenetic protein signaling. J Biol Chem. 281:6203–6210. 2006. View Article : Google Scholar : PubMed/NCBI
4	Lin GL and Hankenson KD: Integration of BMP, Wnt and notch signaling pathways in osteoblast differentiation. J Cell Biochem. 112:3491–3501. 2011. View Article : Google Scholar : PubMed/NCBI
5	Abdallah BM, Jafari A, Zaher W, Qiu W and Kassem M: Skeletal (stromal) stem cells: An update on intracellular signaling pathways controlling osteoblast differentiation. Bone. 70:28–36. 2015. View Article : Google Scholar
6	Kusumbe AP, Ramasamy SK and Adams RH: Coupling of angiogenesis and osteogenesis by a specific vessel subtype in bone. Nature. 507:323–328. 2014. View Article : Google Scholar : PubMed/NCBI
7	Maes C, Carmeliet G and Schipani E: Hypoxia-driven pathways in bone development, regeneration and disease. Nat Rev Rheumatol. 8:358–366. 2012. View Article : Google Scholar : PubMed/NCBI
8	Gustafsson MV, Zheng X, Pereira T, Gradin K, Jin S, Lundkvist J, Ruas JL, Poellinger L, Lendahl U and Bondesson M: Hypoxia requires notch signaling to maintain the undifferentiated cell state. Dev Cell. 9:617–628. 2005. View Article : Google Scholar : PubMed/NCBI
9	Villa JC, Chiu D, Brandes AH, Escorcia FE, Villa CH, Maguire WF, Hu CJ, de Stanchina E, Simon MC, Sisodia SS, et al: Nontranscriptional role of Hif-1α in activation of γ-secretase and notch signaling in breast cancer. Cell Rep. 8:1077–1092. 2014. View Article : Google Scholar : PubMed/NCBI
10	Genetos DC, Toupadakis CA, Raheja LF, Wong A, Papanicolaou SE, Fyhrie DP, Loots GG and Yellowley CE: Hypoxia decreases sclerostin expression and increases Wnt signaling in osteoblasts. J Cell Biochem. 110:457–467. 2010.PubMed/NCBI
11	Chen D, Li Y, Zhou Z, Wu C, Xing Y, Zou X, Tian W and Zhang C: HIF-1α inhibits Wnt signaling pathway by activating Sost expression in osteoblasts. PLoS One. 8:e659402013. View Article : Google Scholar
12	Chen D, Li Y, Zhou Z, Xing Y, Zhong Y, Zou X, Tian W and Zhang C: Synergistic inhibition of Wnt pathway by HIF-1α and osteoblast-specific transcription factor osterix (Osx) in osteoblasts. PLoS One. 7:e529482012. View Article : Google Scholar
13	Hubbi ME and Semenza GL: Regulation of cell proliferation by hypoxia-inducible factors. Am J Physiol Cell Physiol. 309:C775–C782. 2015. View Article : Google Scholar : PubMed/NCBI
14	Braunschweig L, Meyer AK, Wagenführ L and Storch A: Oxygen regulates proliferation of neural stem cells through Wnt/β-catenin signalling. Mol Cell Neurosci. 67:84–92. 2015. View Article : Google Scholar : PubMed/NCBI
15	Meldrum A, Coats P, Orr D and Horgan P: Vascular smooth muscle cell proliferation and signalling pathways – effect of hypoxia on vascular remodelling. Atherosclerosis. 241:e762015. View Article : Google Scholar
16	Canalis E: Wnt signalling in osteoporosis: Mechanisms and novel therapeutic approaches. Nat Rev Endocrinol. 9:575–583. 2013. View Article : Google Scholar : PubMed/NCBI
17	Xu N, Liu H, Qu F, Fan J, Mao K, Yin Y, Liu J, Geng Z and Wang Y: Hypoxia inhibits the differentiation of mesenchymal stem cells into osteoblasts by activation of Notch signaling. Exp Mol Pathol. 94:33–39. 2013. View Article : Google Scholar
18	Chan MC, Holt-Martyn JP, Schofield CJ and Ratcliffe PJ: Pharmacological targeting of the HIF hydroxylases – a new field in medicine development. Mol Aspects Med. Jan 11–2016.Epub ahead of print. View Article : Google Scholar
19	Moriyama H, Moriyama M, Isshi H, Ishihara S, Okura H, Ichinose A, Ozawa T, Matsuyama A and Hayakawa T: Role of notch signaling in the maintenance of human mesenchymal stem cells under hypoxic conditions. Stem Cells Dev. 23:2211–2224. 2014. View Article : Google Scholar : PubMed/NCBI
20	Kim HA, Koo BK, Cho JH, Kim YY, Seong J, Chang HJ, Oh YM, Stange DE, Park JG, Hwang D and Kong YY: Notch1 counteracts WNT/β-catenin signaling through chromatin modification in colorectal cancer. J Clin Invest. 122:3248–3259. 2012. View Article : Google Scholar : PubMed/NCBI
21	Kwon C, Cheng P, King IN, Andersen P, Shenje L, Nigam V and Srivastava D: Notch post-translationally regulates β-catenin protein in stem and progenitor cells. Nat Cell Biol. 13:1244–1251. 2011. View Article : Google Scholar : PubMed/NCBI
22	Andersen P, Uosaki H, Shenje LT and Kwon C: Non-canonical Notch signaling: Emerging role and mechanism. Trends Cell Biol. 22:257–265. 2012. View Article : Google Scholar : PubMed/NCBI
23	Hayward P, Brennan K, Sanders P, Balayo T, DasGupta R, Perrimon N and Martinez Arias A: Notch modulates Wnt signalling by associating with Armadillo/β-catenin and regulating its transcriptional activity. Development. 132:1819–1830. 2005. View Article : Google Scholar : PubMed/NCBI
24	Utting JC, Robins SP, Brandao-Burch A, Orriss IR, Behar J and Arnett TR: Hypoxia inhibits the growth, differentiation and bone-forming capacity of rat osteoblasts. Exp Cell Res. 312:1693–1702. 2006. View Article : Google Scholar : PubMed/NCBI