Osteoadherin serves roles in the regulation of apoptosis and growth in MC3T3‑E1 osteoblast cells

Hamaya,Eri; Fujisawa,Toshiaki; Tamura,Masato

doi:10.3892/ijmm.2019.4376

Osteoadherin serves roles in the regulation of apoptosis and growth in MC3T3‑E1 osteoblast cells

Authors:
- Eri Hamaya
- Toshiaki Fujisawa
- Masato Tamura
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Affiliations: Department of Dental Anesthesiology, Faculty of Dental Medicine and Graduate School of Dental Medicine, Hokkaido University, Sapporo, Hokkaido 060‑8586, Japan, Department of Biochemistry and Molecular Biology, Faculty of Dental Medicine and Graduate School of Dental Medicine, Hokkaido University, Sapporo, Hokkaido 060‑8586, Japan
Published online on: October 18, 2019 https://doi.org/10.3892/ijmm.2019.4376
Pages: 2336-2344

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Abstract

Small leucine‑rich proteoglycans (SLRPs) are a class of proteoglycans that are characterized by small protein cores and structures of leucine‑rich repeats. SLRPs are expressed in most extracellular matrices and share numerous biological functions that are associated with binding of collagens and cell surface receptors. Osteoadherin (also termed osteomodulin) is encoded by the Omd gene and is a keratan sulfate proteoglycan of the class II subfamily of SLRPs. Osteoadherin is highly expressed in mineralized tissues, including bone and dentin; however, it's precise roles remain unknown. The present study determined the Omd expression levels and investigated the effects of over‑ and under‑expression of osteoadherin in osteoblastic cells. Omd mRNA expression increased with osteoblast differentiation in MC3T3‑E1 cells. In C2C12 cells, Omd mRNA expression was induced by bone morphogenetic protein (BMP)2. Reporter assays similarly demonstrated activation of the Omd gene promoter following co‑transfection with Smad1 and Smad4, which are intracellular signaling molecules of the BMP2 signaling pathway. Overexpression of Omd increased the viability and decreased caspase 3/7 activity in MC3T3‑E1 cells. By contrast, following transfection with small interfering RNA for Omd, viable cell numbers were decreased and caspase 3/7 activity was increased. Furthermore, overexpression of Omd reduced the expression of CCN family 2 in these cells. These results demonstrate that Omd expression is regulated during osteoblast differentiation, and that the protein product osteoadherin serves roles in the apoptosis and growth of osteoblast cells.

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1	Heinegård D: Fell-Muir Lecture: Proteoglycans and more-from molecules to biology. Int J Exp Pathol. 90:575–586. 2009. View Article : Google Scholar
2	Iozzo RV and Schaefer L: Proteoglycan form and function: A comprehensive nomenclature of proteoglycans. Matrix Biol. 42:11–55. 2015. View Article : Google Scholar : PubMed/NCBI
3	Wendel M, Sommarin Y and Heinegård D: Bone matrix proteins: Isolation and characterization of a novel cell-binding keratan sulfate proteoglycan (osteoadherin) from bovine bone. J Cell Biol. 141:839–847. 1998. View Article : Google Scholar : PubMed/NCBI
4	Sommarin Y, Wendel M, Shen Z, Hellman U and Heinegârd D: Osteoadherin, a cell-binding keratan sulfate proteoglycan in bone, belongs to the family of leucine-rich repeat proteins of the extracellular matrix. J Biol Chem. 273:16723–16729. 1998. View Article : Google Scholar : PubMed/NCBI
5	Buchaille R, Couble ML, Magloire H and Bleicher F: Expression of the small leucine-rich proteoglycan osteoadherin/osteo-modulin in human dental pulp and developing rat teeth. Bone. 27:265–270. 2000. View Article : Google Scholar : PubMed/NCBI
6	Nikdin H, Olsson ML, Hultenby K and Sugars RV: Osteoadherin accumulates in the predentin towards the mineralization front in the developing tooth. PLoS One. 7:e315252012. View Article : Google Scholar : PubMed/NCBI
7	Sjöberg AP, Manderson GA, Mörgelin M, Day AJ, Heinegård D and Blom AM: Short leucine-rich glycoproteins of the extracellular matrix display diverse patterns of complement interaction and activation. Mol Immunol. 46:830–839. 2009. View Article : Google Scholar :
8	Kalamajski S and Oldberg A: The role of small leucine-rich proteoglycans in collagen fibrillogenesis. Matrix Biol. 29:248–253. 2010. View Article : Google Scholar : PubMed/NCBI
9	Schaefer L and Iozzo RV: Biological functions of the small leucine-rich proteoglycans: From genetics to signal transduction. J Biol Chem. 283:21305–21309. 2008. View Article : Google Scholar : PubMed/NCBI
10	Tillgren V, Onnerfjord P, Haglund L and Heinegård D: The tyrosine sulfate-rich domains of the LRR proteins fibromodulin and osteoadherin bind motifs of basic clusters in a variety of heparin-binding proteins, including bioactive factors. J Biol Chem. 284:28543–28553. 2009. View Article : Google Scholar : PubMed/NCBI
11	Rehn AP, Cerny R, Sugars RV, Kaukua N and Wendel M: Osteoadherin is upregulated by mature osteoblasts and enhances their in vitro differentiation and mineralization. Calcif Tissue Int. 82:454–464. 2008. View Article : Google Scholar : PubMed/NCBI
12	Canalis E, Economides AN and Gazzerro E: Bone morphogenetic proteins, their antagonists, and the skeleton. Endocr Rev. 24:218–235. 2003. View Article : Google Scholar : PubMed/NCBI
13	Chen G, Deng C and Li YP: TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci. 8:272–288. 2012. View Article : Google Scholar :
14	Katagiri T and Tsukamoto S: The unique activity of bone morphogenetic proteins in bone: A critical role of the Smad signaling pathway. Biol Chem. 394:703–714. 2013. View Article : Google Scholar : PubMed/NCBI
15	Van Opdenbosch N and Lamkanfi M: Caspases in cell death, inflammation, and disease. Immunity. 50:1352–1364. 2019. View Article : Google Scholar : PubMed/NCBI
16	Yahara M, Tei K and Tamura M: Inhibition of neuropeptide Y Y1 receptor induces osteoblast differentiation in MC3T3-E1 cells. Mol Med Rep. 16:2779–2784. 2017. View Article : Google Scholar : PubMed/NCBI
17	Tamura M, Sato MM and Nashimoto M: Regulation of CXCL12 expression by canonical Wnt signaling in bone marrow stromal cells. Int J Biochem Cell Biol. 43:760–767. 2011. View Article : Google Scholar : PubMed/NCBI
18	Montesano R, Pepper MS, Möhle-Steinlein U, Risau W, Wagner EF and Orci L: Increased proteolytic activity is responsible for the aberrant morphogenetic behavior of endothelial cells expressing the middle T oncogene. Cell. 62:435–445. 1990. View Article : Google Scholar : PubMed/NCBI
19	Tsuji-Tamura K and Ogawa M: Dual inhibition of mTORC1 and mTORC2 perturbs cytoskeletal organization and impairs endothelial cell elongation. Biochem Biophys Res Commun. 497:326–331. 2018. View Article : Google Scholar : PubMed/NCBI
20	Nakashima A, Katagiri T and Tamura M: Cross-talk between Wnt and bone morphogenetic protein 2 (BMP-2) signaling in differentiation pathway of C2C12 myoblasts. J Biol Chem. 280:37660–37668. 2005. View Article : Google Scholar : PubMed/NCBI
21	Iizuka S, Oridate N, Nashimoto M, Fukuda S and Tamura M: Growth inhibition of head and neck squamous cell carcinoma cells by sgRNA targeting the cyclin D1 mRNA based on TRUE gene silencing. PLoS One. 9:e1141212014. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
23	Nojima J, Kanomata K, Takada Y, Fukuda T, Kokabu S, Ohte S, Takada T, Tsukui T, Yamamoto TS, Sasanuma H, et al: Dual roles of smad proteins in the conversion from myoblasts to osteoblastic cells by bone morphogenetic proteins. J Biol Chem. 285:15577–15586. 2010. View Article : Google Scholar : PubMed/NCBI
24	Teixeira L, Sousa DM, Nunes AF, Sousa MM, Herzog H and Lamghari M: NPY revealed as a critical modulator of osteoblast function in vitro: New insights into the role of Y1 and Y2 receptors. J Cell Biochem. 107:908–916. 2009. View Article : Google Scholar : PubMed/NCBI
25	Khor EC and Baldock P: The NPY system and its neural and neuroendocrine regulation of bone. Curr Osteoporos Rep. 10:160–168. 2012. View Article : Google Scholar : PubMed/NCBI
26	Kurebayashi N, Sato M, Fujisawa T, Fukushima K and Tamura M: Regulation of neuropeptide Y Y1 receptor expression by bone morphogenetic protein 2 in C2C12 myoblasts. Biochem Biophys Res Commun. 439:506–510. 2013. View Article : Google Scholar : PubMed/NCBI
27	Nikitovic D, Aggelidakis J, Young MF, Iozzo RV, Karamanos NK and Tzanakakis GN: The biology of small leucine-rich proteogly-cans in bone pathophysiology. J Biol Chem. 287:33926–33933. 2012. View Article : Google Scholar : PubMed/NCBI
28	Boskey AL, Spevak L, Doty SB and Rosenberg L: Effects of bone CS-proteoglycans, DS-decorin, and DS-biglycan on hydroxyapatite formation in a gelatin gel. Calcif Tissue Int. 61:298–305. 1997. View Article : Google Scholar : PubMed/NCBI
29	Zhou HY: Proteomic analysis of hydroxyapatite interaction proteins in bone. Ann N Y Acad Sci. 1116:323–326. 2007. View Article : Google Scholar : PubMed/NCBI
30	Rehn AP, Chalk AM and Wendel M: Differential regulation of osteoadherin (OSAD) by TGF-beta1 and BMP-2. Biochem Biophys Res Commun. 349:1057–1064. 2006. View Article : Google Scholar : PubMed/NCBI
31	Tang SJ, Hoodless PA, Lu Z, Breitman ML, McInnes RR, Wrana JL and Buchwald M: The Tlx-2 homeobox gene is a downstream target of BMP signalling and is required for mouse mesoderm development. Development. 125:1877–1887. 1998.PubMed/NCBI
32	Dennler S, Itoh S, Vivien D, ten Dijke P, Huet S and Gauthier JM: Direct binding of Smad3 and Smad4 to critical TGF beta-inducible elements in the promoter of human plasminogen activator inhibitor-type 1 gene. EMBO J. 17:3091–3100. 1998. View Article : Google Scholar : PubMed/NCBI
33	Zawel L, Dai JL, Buckhaults P, Zhou S, Kinzler KW, Vogelstein B and Kern SE: Human Smad3 and Smad4 are sequence-specific transcription activators. Mol Cell. 1:611–617. 1998. View Article : Google Scholar : PubMed/NCBI
34	Schaefer L and Schaefer RM: Proteoglycans: From structural compounds to signaling molecules. Cell Tissue Res. 339:237–246. 2010. View Article : Google Scholar
35	Weyers A, Yang B, Solakyildirim K, Yee V, Li L, Zhang F and Linhardt RJ: Isolation of bovine corneal keratan sulfate and its growth factor and morphogen binding. FEBS J. 280:2285–2293. 2013. View Article : Google Scholar : PubMed/NCBI
36	Buraschi S, Neill T, Goyal A, Poluzzi C, Smythies J, Owens RT, Schaefer L, Torres A and Iozzo RV: Decorin causes autophagy in endothelial cells via Peg3. Proc Natl Acad Sci USA. 110:E2582–E2591. 2013. View Article : Google Scholar : PubMed/NCBI
37	Chen CC and Lau LF: Functions and mechanisms of action of CCN matricellular proteins. Int J Biochem Cell Biol. 41:771–783. 2009. View Article : Google Scholar :
38	Kubota S and Takigawa M: Cellular and molecular actions of CCN2/CTGF and its role under physiological and pathological conditions. Clin Sci (Lond). 128:181–196. 2015. View Article : Google Scholar
39	Hishikawa K, Oemar BS, Tanner FC, Nakaki T, Fujii T and Lüscher TF: Overexpression of connective tissue growth factor gene induces apoptosis in human aortic smooth muscle cells. Circulation. 100:2108–2112. 1999. View Article : Google Scholar : PubMed/NCBI
40	Hishikawa K, Oemar BS, Tanner FC, Nakaki T, Lüscher TF and Fujii T: Connective tissue growth factor induces apoptosis in human breast cancer cell line MCF-7. J Biol Chem. 274:37461–37466. 1999. View Article : Google Scholar : PubMed/NCBI
41	Sakai Y, Balam TA, Kuroda S, Tamamura N, Fukunaga T, Takigawa M and Takano-Yamamoto T: CTGF and apoptosis in mouse osteocytes induced by tooth movement. J Dent Res. 88:345–350. 2009. View Article : Google Scholar : PubMed/NCBI