miR‑98‑5p promotes osteoblast differentiation in MC3T3‑E1 cells by targeting CKIP‑1

Liu,Qiliang; Guo,Yong; Wang,Yang; Zou,Xianqiong; Yan,Zhixiong

doi:10.3892/mmr.2018.8416

miR‑98‑5p promotes osteoblast differentiation in MC3T3‑E1 cells by targeting CKIP‑1

Authors:
- Qiliang Liu
- Yong Guo
- Yang Wang
- Xianqiong Zou
- Zhixiong Yan
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Affiliations: Department of Biomedical Engineering, College of Biotechnology of Guilin Medical University, Guilin, Guangxi 541004, P.R. China
Published online on: January 9, 2018 https://doi.org/10.3892/mmr.2018.8416
Pages: 4797-4802

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Abstract

Casein kinase 2-interacting protein 1 (CKIP-1) is a negative regulator for bone formation. Previously, using bioinformatics analysis, CKIP‑1 has been predicted to serve the role of target gene of miR‑98‑5p. In the present study, the potential role of miR‑98‑5p in regulating osteoblast differentiation through CKIP‑1 was investigated. Following pre‑treatment with microRNA (miR)‑98‑5p agomir or miR‑98‑5p antagomir, MC3T3‑E1 cells were cultured in an osteoinductive medium. Subsequently, the expression of miR‑98‑5p, CKIP‑1 and levels of osteoblast differentiation markers, including alkaline phosphatase, matrix mineralization, osteocaicin, collagen type I, runt‑related transcription factor 2 and osteopontin were assayed. Using a dual‑luciferase reporter assay, it was demonstrated that CKIP‑1 was the target gene of miR‑98‑5p. miR‑98‑5p was upregulated as a result of treatment with miR‑98‑5p agomir and promoted osteoblast differentiation. Conversely, miR‑98‑5p antagomir inhibited miR‑98‑5p expression and osteoblast differentiation. miR‑98‑5p targeted CKIP‑1 by binding to its 3'‑untranslated region. Furthermore, miR‑98‑5p overexpression decreased the protein levels of CKIP‑1 and inhibition of miR‑98‑5p increased the protein levels of CKIP‑1. The results of the present study indicated that CKIP‑1 was a target gene of miR‑98‑5p and that miR‑98‑5p regulated osteoblast differentiation in MC3T3‑E1 cells by targeting CKIP‑1.

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1	Lerner UH: Osteoblasts, osteoclasts, and osteocytes: Unveiling their intimate associated responses to applied orthodontic forces. Semin in Orthodon. 18:237–248. 2012. View Article : Google Scholar
2	Rolfe R, Roddy K and Murphy P: Mechanical regulation of skeletal development. Curr Osteoporos Rep. 11:107–116. 2013. View Article : Google Scholar : PubMed/NCBI
3	Lu K, Yin X, Weng T, Xi S, Li L, Xing G, Cheng X, Yang X, Zhang L and He F: Targeting WW domains linker of HECT-type ubiquitin ligase Smurf1 for activation by CKIP-1. Nat Cell Biol. 10:994–1002. 2008. View Article : Google Scholar : PubMed/NCBI
4	Nie J, Liu L, He F, Fu X, Han W and Zhang L: CKIP-1: A scaffold protein and potential therapeutic target integrating multiple signaling pathways and physiological functions. Ageing Res Rev. 12:276–281. 2013. View Article : Google Scholar : PubMed/NCBI
5	Zhang G, Guo B, Wu H, Tang T, Zhang BT, Zheng L, He Y, Yang Z, Pan X, Chow H, et al: A delivery system targeting bone formation surfaces to facilitate RNAi-based anabolic therapy. Nat Med. 18:307–314. 2012. View Article : Google Scholar : PubMed/NCBI
6	Guo B, Zhang B, Zheng L, Tang T, Liu J, Wu H, Yang Z, Peng S, He X, Zhang H, et al: Therapeutic RNA interference targeting CKIP-1 with a cross-species sequence to stimulate bone formation. Bone. 59:76–88. 2014. View Article : Google Scholar : PubMed/NCBI
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
8	Winter J, Jung S, Keller S, Gregory RI and Diederichs S: Many roads to maturity: microRNA biogenesis pathways and their regulation. Nat Cell Biol. 11:228–234. 2009. View Article : Google Scholar : PubMed/NCBI
9	Mendell JT and Olson EN: MicroRNAs in stress signaling and human disease. Cell. 148:1172–1187. 2012. View Article : Google Scholar : PubMed/NCBI
10	Ranganathan K and Sivasankar V: MicroRNAs-biology and clinical applications. J Oral Maxillofac Pathol. 18:229–234. 2014. View Article : Google Scholar : PubMed/NCBI
11	Vimalraj S and Selvamurugan N: MicroRNAs: Synthesis, gene regulation and osteoblast differentiation. Curr Issues Mol Biol. 15:7–18. 2013.PubMed/NCBI
12	Huang J, Zhao L, Xing L and Chen D: MicroRNA-204 regulates Runx2 protein expression and mesenchymal progenitor cell differentiation. Stem Cells. 28:357–364. 2010.PubMed/NCBI
13	Hassan MQ, Maeda Y, Taipaleenmaki H, Zhang W, Jafferji M, Gordon JA, Li Z, Croce CM, van Wijnen AJ, Stein JL, et al: miR-218 directs a Wnt signaling circuit to promote differentiation of osteoblasts and osteomimicry of metastatic cancer cells. J Biol Chem. 287:42084–42092. 2012. View Article : Google Scholar : PubMed/NCBI
14	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
15	Beck GR Jr, Zerler B and Moran E: Phosphate is a specific signal for induction of osteopontin gene expression. Proc Natl Acad Sci USA. 97:pp. 8352–8357. 2000; View Article : Google Scholar : PubMed/NCBI
16	Wu M, Hesse E, Morvan F, Zhang JP, Correa D, Rowe GC, Kiviranta R, Neff L, Philbrick WM, Horne WC and Baron R: Zfp521 antagonizes Runx2, delays osteoblast differentiation in vitro and promotes bone formation in vivo. Bone. 44:528–536. 2009. View Article : Google Scholar : PubMed/NCBI
17	Mahalingam CD, Datta T, Patil RV, Kreider J, Bonfil RD, Kirkwood KL, Goldstein SA, Abou-Samra AB and Datta NS: Mitogen-activated protein kinase phosphatase 1 regulates bone mass, osteoblast gene expression, and responsiveness to parathyroid hormone. J Endocrinol. 211:145–156. 2011. View Article : Google Scholar : PubMed/NCBI
18	Guo Y, Liu L, Hao Q, Li R, Zhang X, Wang L and Ning B: Effects of extracellular matrix produced in vitro on growth and differentiation of MC3T3-E1 cells. Sheng Wu Gong Cheng Xue Bao. 27:1606–1612. 2011.PubMed/NCBI
19	Guo Y, Zhang CQ, Zeng QC, Li RX, Liu L, Hao QX, Shi CH, Zhang XZ and Yan YX: Mechanical strain promotes osteoblast ECM formation and improves its osteoinductive potential. Biomed Eng Online. 11:802012. View Article : Google Scholar : PubMed/NCBI
20	Bhatt KA, Chang EI, Warren SM, Lin SE, Bastidas N, Ghali S, Thibboneir A, Capla JM, McCarthy JG and Gurtner GC: Uniaxial mechanical strain: An in vitro correlate to distraction osteogenesis. J Surg Res. 143:329–336. 2007. View Article : Google Scholar : PubMed/NCBI
21	Hu G, Zhou R, Liu J, Gong AY, Eischeid AN, Dittman JW and Chen XM: MicroRNA-98 and let-7 confer cholangiocyte expression of cytokine-inducible Src homology 2-containing protein in response to microbial challenge. J Immunol. 183:1617–1624. 2009. View Article : Google Scholar : PubMed/NCBI
22	Liu Y, Chen Q, Song Y, Lai L, Wang J, Yu H, Cao X and Wang Q: MicroRNA-98 negatively regulates IL-10 production and endotoxin tolerance in macrophages after LPS stimulation. FEBS Lett. 585:1963–1968. 2011. View Article : Google Scholar : PubMed/NCBI
23	Ting HJ, Messing J, Yasmin-Karim S and Lee YF: Identification of microRNA-98 as a therapeutic target inhibiting prostate cancer growth and a biomarker induced by vitamin D. J Biol Chem. 288:1–9. 2013. View Article : Google Scholar : PubMed/NCBI
24	Liu T, Hou L and Huang Y: EZH2-specific microRNA-98 inhibits human ovarian cancer stem cell proliferation via regulating the pRb-E2F pathway. Tumour Biol. 35:7239–7247. 2014. View Article : Google Scholar : PubMed/NCBI
25	Zhou W, Zou B, Liu L, Cui K, Gao J, Yuan S and Cong N: MicroRNA-98 acts as a tumor suppressor in hepatocellular carcinoma via targeting SALL4. Oncotarget. 7:74059–74073. 2016. View Article : Google Scholar : PubMed/NCBI
26	Wang CY, Zhang JJ, Hua L, Yao KH, Chen JT and Ren XQ: MicroRNA-98 suppresses cell proliferation, migration and invasion by targeting collagen triple helix repeat containing 1 in hepatocellular carcinoma. Mol Med Rep. 13:2639–2644. 2016. View Article : Google Scholar : PubMed/NCBI
27	Zhu X, Ouyang Y, Zhong F, Wang Q, Ding L, Zhang P, Chen L, Liu H and He S: Silencing of CKIP-1 promotes tumor proliferation and cell adhesion-mediated drug resistance via regulating AKT activity in non-Hodgkin's lymphoma. Oncol Rep. 37:622–630. 2017. View Article : Google Scholar : PubMed/NCBI
28	Zhang L, Wang Y, Xiao F, Wang S, Xing G, Li Y, Yin X, Lu K, Wei R, Fan J, et al: CKIP-1 regulates macrophage proliferation by inhibiting TRAF6-mediated Akt activation. Cell Res. 24:742–761. 2014. View Article : Google Scholar : PubMed/NCBI
29	Zhang X, Wang Q, Wan Z, Li J, Liu L and Zhang X: CKIP-1 knockout offsets osteoporosis induced by simulated microgravity. Prog Biophys Mol Biol. 122:140–148. 2016. View Article : Google Scholar : PubMed/NCBI
30	Zhou ZC, Che L, Kong L, Lei DL, Liu R and Yang XJ: CKIP-1 silencing promotes new bone formation in rat mandibular distraction osteogenesis. Oral Surg Oral Med Oral Pathol Oral Radiol. 123:e1–e9. 2017. View Article : Google Scholar : PubMed/NCBI