miR‑217 inhibits osteogenic differentiation of rat bone marrow‑derived mesenchymal stem cells by binding to Runx2

Zhu,Yu‑Long; Wang,Shui; Ding,De‑Gang; Xu,Liang; Zhu,Hai‑Tao

doi:10.3892/mmr.2017.6349

miR‑217 inhibits osteogenic differentiation of rat bone marrow‑derived mesenchymal stem cells by binding to Runx2

Authors:
- Yu‑Long Zhu
- Shui Wang
- De‑Gang Ding
- Liang Xu
- Hai‑Tao Zhu
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Affiliations: Department of Orthopedics, Sheyang County People's Hospital, Yancheng, Jiangsu 224300, P.R. China
Published online on: March 22, 2017 https://doi.org/10.3892/mmr.2017.6349
Pages: 3271-3277

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Abstract

The elucidation of the underlying molecular mechanisms regulating the osteogenic differentiation of bone marrow‑derived mesenchymal stem cells (BMSCs) is of great importance in improving the treatment of bone‑associated diseases. MicroRNAs (miRNAs) have been proven to regulate the osteogenic differentiation of BMSCs. The present study investigated the role of miR‑217 in the osteogenic differentiation of rat BMSCs. It was observed that miR‑217 expression levels were downregulated during the process of osteogenic differentiation. Subsequently, a dual‑luciferase reporter gene assay demonstrated that miR‑217 targets a putative binding site in the 3'‑untranslated region of the runt related transcription factor 2 (Runx2) gene, which is a key transcription factor for osteogenesis. It was then demonstrated that overexpression of miR‑217 attenuated the osteogenesis of BMSCs and downregulated the expression of Runx2, whereas inhibition of miR‑217 promoted osteoblastic differentiation and upregulated Runx2 expression. Furthermore, the extracellular signal‑regulated kinase (ERK) and p38 mitogen‑activated protein kinase (p38 MAPK) signaling pathways were investigated during osteogenic induction, and the data indicated that miR‑217 may exert a negative effect on the osteogenic differentiation of BMSCs through alteration of ERK and p38 MAPK phosphorylation. The present study therefore concluded that miR‑217 functions as a negative regulator of BMSC osteogenic differentiation via the inhibition of Runx2 expression, and the underlying molecular mechanisms may partially be attributed to mediation by the ERK and p38 MAPK signaling pathways.

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1	Corral DA, Amling M, Priemel M, Loyer E, Fuchs S, Ducy P, Baron R and Karsenty G: Dissociation between bone resorption and bone formation in osteopenic transgenic mice. Proc Natl Acad Sci USA. 95:13835–13840. 1998. View Article : Google Scholar : PubMed/NCBI
2	Charbord P: Bone marrow mesenchymal stem cells: Historical overview and concepts. Hum Gene Ther. 21:1045–1056. 2010. View Article : Google Scholar : PubMed/NCBI
3	Sacchetti B, Funari A, Michienzi S, Di Cesare S, Piersanti S, Saggio I, Tagliafico E, Ferrari S, Robey PG, Riminucci M and Bianco P: Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell. 131:324–336. 2007. View Article : Google Scholar : PubMed/NCBI
4	Mathews S, Bhonde R, Gupta PK and Totey S: Extracellular matrix protein mediated regulation of the osteoblast differentiation of bone marrow derived human mesenchymal stem cells. Differentiation. 84:185–192. 2012. View Article : Google Scholar : PubMed/NCBI
5	Khosla S, Westendorf JJ and Oursler MJ: Building bone to reverse osteoporosis and repair fractures. J Clin Invest. 118:421–428. 2008. View Article : Google Scholar : PubMed/NCBI
6	Ghildiyal M and Zamore PD: Small silencing RNAs: An expanding universe. Nat Rev Genet. 10:94–108. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ha M and Kim VN: Regulation of microRNA biogenesis. Nat Rev Mol Cell Biol. 15:509–524. 2014. View Article : Google Scholar : PubMed/NCBI
8	Wu T, Zhou H, Hong Y, Li J, Jiang X and Huang H: miR-30 family members negatively regulate osteoblast differentiation. J Biol Chem. 287:7503–7511. 2012. View Article : Google Scholar : PubMed/NCBI
9	Kim KM, Park SJ, Jung SH, Kim EJ, Jogeswar G, Ajita J, Rhee Y, Kim CH and Lim SK: miR-182 is a negative regulator of osteoblast proliferation, differentiation, and skeletogenesis through targeting FoxO1. J Bone Miner Res. 27:1669–1679. 2012. View Article : Google Scholar : PubMed/NCBI
10	Wang Q, Cai J, Cai XH and Chen L: miR-346 regulates osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting the Wnt/β-catenin pathway. PLoS One. 8:e722662013. View Article : Google Scholar : PubMed/NCBI
11	Wei R, Deng Z and Su J: miR-217 targeting Wnt5a in osteosarcoma functions as a potential tumor suppressor. Biomed Pharmacother. 72:158–164. 2015. View Article : Google Scholar : PubMed/NCBI
12	Chen DL, Zhang DS, Lu YX, Chen LZ, Zeng ZL, He MM, Wang FH, Li YH, Zhang HZ, Pelicano H, et al: microRNA-217 inhibits tumor progression and metastasis by downregulating EZH2 and predicts favorable prognosis in gastric cancer. Oncotarget. 6:10868–10879. 2015. View Article : Google Scholar : PubMed/NCBI
13	Zhao WG, Yu SN, Lu ZH, Ma YH, Gu YM and Chen J: The miR-217 microRNA functions as a potential tumor suppressor in pancreatic ductal adenocarcinoma by targeting KRAS. Carcinogenesis. 31:1726–1733. 2010. View Article : Google Scholar : PubMed/NCBI
14	Li H, Zhao J, Zhang JW, Huang QY, Huang JZ, Chi LS, Tang HJ, Liu GQ, Zhu DJ and Ma WM: MicroRNA-217, down-regulated in clear cell renal cell carcinoma and associated with lower survival, suppresses cell proliferation and migration. Neoplasma. 60:511–515. 2013. View Article : Google Scholar : PubMed/NCBI
15	Pan GZ, Yang Y, Zhang J, Liu W, Wang GY, Zhang YC, Yang Q, Zhai FX, Tai Y, Liu JR, et al: Bone marrow mesenchymal stem cells ameliorate hepatic ischemia/reperfusion injuries via inactivation of the MEK/ERK signaling pathway in rats. J Surg Res. 178:935–948. 2012. View Article : Google Scholar : PubMed/NCBI
16	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 : PubMed/NCBI
17	Qu B, Xia X, Wu HH, Tu CQ and Pan XM: PDGF-regulated miRNA-138 inhibits the osteogenic differentiation of mesenchymal stem cells. Biochem Biophys Res Commun. 448:241–247. 2014. View Article : Google Scholar : PubMed/NCBI
18	Xu L, Liu Y, Hou Y, Wang K, Wong Y, Lin S and Li G: U0126 promotes osteogenesis of rat bone-marrow-derived mesenchymal stem cells by activating BMP/Smad signaling pathway. Cell Tissue Res. 359:537–545. 2015. View Article : Google Scholar : PubMed/NCBI
19	Lewis BP, Shih IH, Jones-Rhoades MW, Bartel DP and Burge CB: Prediction of mammalian microRNA targets. Cell. 115:787–798. 2003. View Article : Google Scholar : PubMed/NCBI
20	Krzeszinski JY, Wei W, Huynh H, Jin Z, Wang X, Chang TC, Xie XJ, He L, Mangala LS, Lopez-Berestein G, et al: miR-34a blocks osteoporosis and bone metastasis by inhibiting osteoclastogenesis and Tgif2. Nature. 512:431–435. 2014. View Article : Google Scholar : PubMed/NCBI
21	Garnero P: New developments in biological markers of bone metabolism in osteoporosis. Bone. 66:46–55. 2014. View Article : Google Scholar : PubMed/NCBI
22	Qadir AS, Um S, Lee H, Baek K, Seo BM, Lee G, Kim GS, Woo KM, Ryoo HM and Baek JH: miR-124 negatively regulates osteogenic differentiation and in vivo bone formation of mesenchymal stem cells. J Cell Biochem. 116:730–742. 2015. View Article : Google Scholar : PubMed/NCBI
23	Huszar JM and Payne CJ: MIR146A inhibits JMJD3 expression and osteogenic differentiation in human mesenchymal stem cells. FEBS Lett. 588:1850–1856. 2014. View Article : Google Scholar : PubMed/NCBI
24	Huang K, Fu J, Zhou W, Li W, Dong S, Yu S, Hu Z, Wang H and Xie Z: MicroRNA-125b regulates osteogenic differentiation of mesenchymal stem cells by targeting Cbfβ in vitro. Biochimie. 102:47–55. 2014. View Article : Google Scholar : PubMed/NCBI
25	Deng Y, Wu S, Zhou H, Bi X, Wang Y, Hu Y, Gu P and Fan X: Effects of a miR-31, Runx2, and Satb2 regulatory loop on the osteogenic differentiation of bone mesenchymal stem cells. Stem Cells Dev. 22:2278–2286. 2013. View Article : Google Scholar : PubMed/NCBI
26	Komori T, Yagi H, Nomura S, Yamaguchi A, Sasaki K, Deguchi K, Shimizu Y, Bronson RT, Gao YH, Inada M, et al: Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. Cell. 89:755–764. 1997. View Article : Google Scholar : PubMed/NCBI
27	Yoshida CA, Furuichi T, Fujita T, Fukuyama R, Kanatani N, Kobayashi S, Satake M, Takada K and Komori T: Core-binding factor beta interacts with Runx2 and is required for skeletal development. Nat Genet. 32:633–638. 2002. View Article : Google Scholar : PubMed/NCBI
28	Lian JB and Stein GS: Runx2/Cbfa1: A multifunctional regulator of bone formation. Curr Pharm Des. 9:2677–2685. 2003. View Article : Google Scholar : PubMed/NCBI
29	Lai CF, Chaudhary L, Fausto A, Halstead LR, Ory DS, Avioli LV and Cheng SL: Erk is essential for growth, differentiation, integrin expression, and cell function in human osteoblastic cells. J Biol Chem. 276:14443–14450. 2001.PubMed/NCBI
30	Simmons CA, Matlis S, Thornton AJ, Chen S, Wang CY and Mooney DJ: Cyclic strain enhances matrix mineralization by adult human mesenchymal stem cells via the extracellular signal-regulated kinase (ERK1/2) signaling pathway. J Biomech. 36:1087–1096. 2003. View Article : Google Scholar : PubMed/NCBI
31	Xiao G, Jiang D, Thomas P, Benson MD, Guan K, Karsenty G and Franceschi RT: MAPK pathways activate and phosphorylate the osteoblast-specific transcription factor, Cbfa1. J Biol Chem. 275:4453–4459. 2000. View Article : Google Scholar : PubMed/NCBI