Role of the Wnt/β-catenin signaling pathway in the response of chondrocytes to mechanical loading

Niu,Qiannan; Li,Feifei; Zhang,Liang; Xu,Xinyuan; Liu,Yucong; Gao,Jie; Feng,Xue

doi:10.3892/ijmm.2016.2463

Role of the Wnt/β-catenin signaling pathway in the response of chondrocytes to mechanical loading

Authors:
- Qiannan Niu
- Feifei Li
- Liang Zhang
- Xinyuan Xu
- Yucong Liu
- Jie Gao
- Xue Feng
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Affiliations: State Key Laboratory of Military Stomatology, Department of Orthodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Stomatology, Hospital 323 of The People's Liberation Army, Xi'an, Shaanxi 710045, P.R. China, Department of Biochemistry and Molecular Biology, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi 710032, P.R. China, Department of Stomatology, The First People's Hospital of Shuangliu County, Chengdu, Sichuan 610200, P.R. China
Published online on: January 22, 2016 https://doi.org/10.3892/ijmm.2016.2463
Pages: 755-762

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Abstract

In order to better understand the mechanisms by which chondrocytes respond to mechanical stimulation, ATDC5 mouse embryonic carcinoma cells were induced to differentiate into chondrocytes and then exposed to mechanical loading. To specifically elucidate the role of this pathway, the localization and expression of proteins involved in the Wnt/β-catenin signaling pathway were observed. Chondrogenic-differentiated ATDC5 cells were exposed to a 12% cycle tension load for 1, 2, 4, or 8 h. At each time point, immunofluorescence staining, western blot analysis, and qPCR were used to track the localization of β-catenin and glycogen synthase kinase-3β (GSK-3β) expression. In addition, the mRNA expression of Wnt3a, disheveled homolog 1 (Dvl-1), GSK-3β, and collagen type II were also detected. Activation of the Wnt/β-catenin signaling pathway was investigated in cells treated with Dickkopf-related protein 1 (DKK-1). β-catenin and GSK-3β protein expression increased initially and then decreased over the mechanical loading period, and the corresponding mRNA levels followed a similar trend. After application of the inhibitor DKK-1, Wnt/β‑catenin signaling was suppressed, and the mRNA expression of collagen II was also reduced. Thus, stimulation of chondrocytes with mechanical strain loading is associated with the translocation of active β-catenin from the cytoplasm to the nucleus.

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1	Kinzinger G, Kober C and Diedrich P: Topography and morphology of the mandibular condyle during fixed functional orthopedic treatment - a magnetic resonance imaging study. J Orofac Orthop. 68:124–147. 2007. View Article : Google Scholar : PubMed/NCBI
2	Bewyer DC: Biomechanical and physiologic processes leading to internal derangement with adhesion. J Craniomandib Disord. 3:44–49. 1989.PubMed/NCBI
3	Meikle MC: Remodeling the dentofacial skeleton: the biological basis for orthodontics and dentofacial orthopedics. J Dent Res. 86:12–24. 2007. View Article : Google Scholar
4	Habib H, Hatta T, Udagawa J, Zhang L, Yoshimura Y and Otani H: Fetal jaw movement affects condylar cartilage development. J Dent Res. 84:474–479. 2005. View Article : Google Scholar : PubMed/NCBI
5	Madhavan S, Anghelina M, Rath-Deschner B, Wypasek E, John A, Deschner J, Piesco N and Agarwal S: Biomechanical signals exert sustained attenuation of proinflammatory gene induction in articular chondrocytes. Osteoarthritis Cartilage. 14:1023–1032. 2006. View Article : Google Scholar : PubMed/NCBI
6	Ker RF, Wang XT and Pike AV: Fatigue quality of mammalian tendons. J Exp Biol. 203:1317–1327. 2000.PubMed/NCBI
7	Kuboki T, Shinoda M, Orsini MG and Yamashita A: Viscoelastic properties of the pig temporomandibular joint articular soft tissues of the condyle and disc. J Dent Res. 76:1760–1769. 1997. View Article : Google Scholar : PubMed/NCBI
8	Kronenberg HM: Developmental regulation of the growth plate. Nature. 423:332–336. 2003. View Article : Google Scholar : PubMed/NCBI
9	Maruyama T, Mirando AJ, Deng CX and Hsu W: The balance of WNT and FGF signaling influences mesenchymal stem cell fate during skeletal development. Sci Signal. 3:ra402010.PubMed/NCBI
10	Larsson T, Aspden RM and Heinegård D: Effects of mechanical load on cartilage matrix biosynthesis in vitro. Matrix. 11:388–394. 1991. View Article : Google Scholar : PubMed/NCBI
11	Church V, Nohno T, Linker C, Marcelle C and Francis-West P: Wnt regulation of chondrocyte differentiation. J Cell Sci. 115:4809–4818. 2002. View Article : Google Scholar : PubMed/NCBI
12	Chimal-Monroy J, Montero JA, Gañan Y, Macias D, Garcia-Porrero JA and Hurle JM: Comparative analysis of the expression and regulation of Wnt5a, Fz4, and Frzb1 during digit formation and in micromass cultures. Dev Dyn. 224:314–320. 2002. View Article : Google Scholar : PubMed/NCBI
13	Atsumi T, Miwa Y, Kimata K and Ikawa Y: A chondrogenic cell line derived from a differentiating culture of AT805 teratocarcinoma cells. Cell Differ Dev. 30:109–116. 1990. View Article : Google Scholar : PubMed/NCBI
14	Shukunami C, Shigeno C, Atsumi T, Ishizeki K, Suzuki F and Hiraki Y: Chondrogenic differentiation of clonal mouse embryonic cell line ATDC5 in vitro: differentiation-dependent gene expression of parathyroid hormone (PTH)/PTH-related peptide receptor. J Cell Biol. 133:457–468. 1996. View Article : Google Scholar : PubMed/NCBI
15	Saito-Diaz K, Chen TW, Wang X, Thorne CA, Wallace HA, Page-McCaw A and Lee E: The way Wnt works: components and mechanism. Growth Factors. 31:1–31. 2013. View Article : Google Scholar :
16	Gould TD and Manji HK: Glycogen synthase kinase-3: a putative molecular target for lithium mimetic drugs. Neuropsychopharmacology. 30:1223–1237. 2005.PubMed/NCBI
17	Case N, Ma M, Sen B, Xie Z, Gross TS and Rubin J: Beta-catenin levels influence rapid mechanical responses in osteoblasts. J Biol Chem. 283:29196–29205. 2008. View Article : Google Scholar : PubMed/NCBI
18	Liu F, Kohlmeier S and Wang CY: Wnt signaling and skeletal development. Cell Signal. 20:999–1009. 2008. View Article : Google Scholar : PubMed/NCBI
19	Aguilera O, Fraga MF, Ballestar E, Paz MF, Herranz M, Espada J, García JM, Muñoz A, Esteller M and González-Sancho JM: Epigenetic inactivation of the Wnt antagonist DICKKOPF-1 (DKK-1) gene in human colorectal cancer. Oncogene. 25:4116–4121. 2006. View Article : Google Scholar : PubMed/NCBI
20	González-Sancho JM, Aguilera O, García JM, Pendás-Franco N, Peña C, Cal S, García de Herreros A, Bonilla F and Muñoz A: The Wnt antagonist DICKKOPF-1 gene is a downstream target of beta-catenin/TCF and is downregulated in human colon cancer. Oncogene. 24:1098–1103. 2005. View Article : Google Scholar
21	Alanen P: Occlusion and temporomandibular disorders (TMD): still unsolved question? J Dent Res. 81:518–519. 2002. View Article : Google Scholar : PubMed/NCBI
22	Hartmann C and Tabin CJ: Dual roles of Wnt signaling during chondrogenesis in the chicken limb. Development. 127:3141–3159. 2000.PubMed/NCBI
23	Rudnicki JA and Brown AM: Inhibition of chondrogenesis by Wnt gene expression in vivo and in vitro. Dev Biol. 185:104–118. 1997. View Article : Google Scholar : PubMed/NCBI
24	Yano F, Kugimiya F, Ohba S, Ikeda T, Chikuda H, Ogasawara T, Ogata N, Takato T, Nakamura K, Kawaguchi H and Chung UI: The canonical Wnt signaling pathway promotes chondrocyte differentiation in a Sox9-dependent manner. Biochem Biophys Res Commun. 333:1300–1308. 2005. View Article : Google Scholar : PubMed/NCBI
25	Brannon M, Gomperts M, Sumoy L, Moon RT and Kimelman D: A beta-catenin/XTcf-3 complex binds to the siamois promoter to regulate dorsal axis specification in Xenopus. Genes Dev. 11:2359–2370. 1997. View Article : Google Scholar : PubMed/NCBI
26	van de Wetering M, Cavallo R, Dooijes D, van Beest M, van Es J, Loureiro J, Ypma A, Hursh D, Jones T, Bejsovec A, et al: Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF. Cell. 88:789–799. 1997. View Article : Google Scholar : PubMed/NCBI
27	Ratcliffe A, Billingham ME, Saed-Nejad F, Muir H and Hardingham TE: Increased release of matrix components from articular cartilage in experimental canine osteoarthritis. J Orthop Res. 10:350–358. 1992. View Article : Google Scholar : PubMed/NCBI
28	Krupnik VE, Sharp JD, Jiang C, Robison K, Chickering TW, Amaravadi L, Brown DE, Guyot D, Mays G, Leiby K, et al: Functional and structural diversity of the human Dickkopf gene family. Gene. 238:301–313. 1999. View Article : Google Scholar : PubMed/NCBI
29	Premaraj S, Souza I and Premaraj T: Mechanical loading activates β-catenin signaling in periodontal ligament cells. Angle Orthod. 81:592–599. 2011. View Article : Google Scholar : PubMed/NCBI
30	Rizkalla G, Reiner A, Bogoch E and Poole AR: Studies of the articular cartilage proteoglycan aggrecan in health and osteoarthritis. Evidence for molecular heterogeneity and extensive molecular changes in disease. J Clin Invest. 90:2268–2277. 1992. View Article : Google Scholar : PubMed/NCBI