miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β‑catenin signaling

Cao,Fengdi; Zhan,Jialin; Chen,Xufeng; Zhang,Kai; Lai,Renfa; Feng,Zhiqiang

doi:10.3892/mmr.2017.7821

miR-214 promotes periodontal ligament stem cell osteoblastic differentiation by modulating Wnt/β‑catenin signaling

Authors:
- Fengdi Cao
- Jialin Zhan
- Xufeng Chen
- Kai Zhang
- Renfa Lai
- Zhiqiang Feng
View Affiliations

Affiliations: Department of Stomatology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510630, P.R. China, Department of Ophthalmology, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Tianjin Key Laboratory of Ionic‑Molecular Function of Cardiovascular Disease, Department of Cardiology, Second Hospital of Tianjin Medical University, Tianjin Institute of Cardiology, Tianjin 300211, P.R. China
Published online on: October 19, 2017 https://doi.org/10.3892/mmr.2017.7821
Pages: 9301-9308
Copyright: © Cao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The canonical Wnt/β‑catenin signaling is important in the differentiation of human mesenchymal stem cells into osteoblasts. Accumulating evidence suggests that the expression of β‑catenin is, in part, regulated by specific microRNAs (miRNAs). The aim of the present study was to investigate the putative roles of miRNAs in osteoblast differentiation. Polymerase chain reaction (PCR) arrays were used to identify miRNAs that were differentially expressed between differentiated and non‑differentiated periodontal ligament stem cells (PDLSCs), and reverse transcription‑quantitative PCR (RT‑qPCR) was used for validation. Since miR‑214 was revealed to be significantly downregulated during PDLSC differentiation, its function was further investigated via silencing and overexpression. In addition, osteogenic differentiation of PDLSCs was evaluated at 10 and 21 days following induction, using Alizarin red staining and RT‑qPCR analysis for mRNA expression levels of the osteogenic differentiation markers alkaline phosphatase (ALP), osteocalcin and bone sialoprotein. Furthermore, the potential target genes of miR‑214 were investigated using a dual‑luciferase reporter assay, RT‑qPCR and western blot analysis, whereas a TOPflash/FOPflash reporter plasmid system followed by a luciferase assay was used to examine the effects of miR‑214 on Wnt/β‑catenin signaling. The present results demonstrated that miR‑214 was significantly downregulated during the osteoblastic differentiation of PDLSCs. Notably, its overexpression inhibited PDLSC differentiation, whereas its knockdown promoted PDLSC differentiation, as revealed by alterations in mRNA expression of osteoblast‑specific genes and ALP. In addition, miR‑214 was demonstrated to directly interact with the 3'‑untranslated region of the β‑catenin gene CTNNB1, and suppressed Wnt/β‑catenin signaling through the inhibition of β‑catenin. The results of the present study suggested that miR‑214 may participate in the regulation of the Wnt/β‑catenin signaling pathway, and may have potential as a candidate target for the development of preventive or therapeutic agents for the treatment of patients with osteogenic disorders.

View Figures

View References

1	Armitage GC: Periodontal diagnoses and classification of periodontal diseases. Periodontol. 34:9–21. 2000. View Article : Google Scholar
2	Hiraga T, Ninomiya T, Hosoya A, Takahashi M and Nakamura H: Formation of bone-like mineralized matrix by periodontal ligament cells in vivo: A morphological study in rats. J Bone Miner Metab. 27:149–157. 2009. View Article : Google Scholar : PubMed/NCBI
3	Huang GT, Gronthos S and Shi S: Mesenchymal stem cells derived from dental tissues vs. those from other sources: Their biology and role in regenerative medicine. J Dent Res. 88:792–806. 2009. View Article : Google Scholar : PubMed/NCBI
4	Kato T, Hattori K, Deguchi T, Katsube Y, Matsumoto T, Ohgushi H and Numabe Y: Osteogenic potential of rat stromal cells derived from periodontal ligament. J Tissue Eng Regen Med. 5:798–805. 2011. View Article : Google Scholar : PubMed/NCBI
5	Lamplot JD, Qin J, Nan G, Wang J, Liu X, Yin L, Tomal J, Li R, Shui W, Zhang H, et al: BMP9 signaling in stem cell differentiation and osteogenesis. Am J Stem Cells. 2:1–21. 2013.PubMed/NCBI
6	Coleman DT, Gray AL, Stephens CA, Scott ML and Cardelli JA: Repurposed drug screen identifies cardiac glycosides as inhibitors of TGF-β-induced cancer-associated fibroblast differentiation. Oncotarget. 7:32200–32209. 2016. View Article : Google Scholar : PubMed/NCBI
7	Ishitani T, Kishida S, Hyodo-Miura J, Ueno N, Yasuda J, Waterman M, Shibuya H, Moon RT, Ninomiya-Tsuji J and Matsumoto K: The TAK1-NLK mitogen-activated protein kinase cascade functions in the Wnt-5a/Ca(2+) pathway to antagonize Wnt/beta-catenin signaling. Mol Cell Biol. 23:131–139. 2003. View Article : Google Scholar : PubMed/NCBI
8	Boland GM, Perkins G, Hall DJ and Tuan RS: Wnt 3a promotes proliferation and suppresses osteogenic differentiation of adult human mesenchymal stem cells. J Cell Biochem. 93:1210–1230. 2004. View Article : Google Scholar : PubMed/NCBI
9	Hartmann C and Tabin CJ: Dual roles of Wnt signaling during chondrogenesis in the chicken limb. Development. 127:3141–3159. 2000.PubMed/NCBI
10	Ross SE, Hemati N, Longo KA, Bennett CN, Lucas PC, Erickson RL and MacDougald OA: Inhibition of adipogenesis by Wnt signaling. Science. 289:950–953. 2000. View Article : Google Scholar : PubMed/NCBI
11	Shang Y, Zhang C, Wang S, Xiong F, Zhao C, Peng F, Feng S, Yu M, Li M and Zhang Y: Activated beta-catenin induces myogenesis and inhibits adipogenesis in BM-derived mesenchymal stromal cells. Cytotherapy. 9:667–681. 2007. View Article : Google Scholar : PubMed/NCBI
12	Mizuno Y, Yagi K, Tokuzawa Y, Kanesaki-Yatsuka Y, Suda T, Katagiri T, Fukuda T, Maruyama M, Okuda A, Amemiya T, et al: miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun. 368:267–272. 2008. View Article : Google Scholar : PubMed/NCBI
13	Liang H, Li X, Wang L, Yu S, Xu Z, Gu Y, Pan Z, Li T, Hu M, Cui H, et al: MicroRNAs contribute to promyelocyte apoptosis in As2O3-treated APL cells. Cell Physiol Biochem. 32:1818–1829. 2013. View Article : Google Scholar : PubMed/NCBI
14	Feng J, Iwama A, Satake M and Kohu K: MicroRNA-27 enhances differentiation of myeloblasts into granulocytes by post-transcriptionally downregulating Runx1. Br J Haematol. 145:412–423. 2009. View Article : Google Scholar : PubMed/NCBI
15	McDaneld TG, Smith TP, Doumit ME, Miles JR, Coutinho LL, Sonstegard TS, Matukumalli LK, Nonneman DJ and Wiedmann RT: MicroRNA transcriptome profiles during swine skeletal muscle development. BMC Genomics. 10:772009. View Article : Google Scholar : PubMed/NCBI
16	Zhou Q, Zhao ZN, Cheng JT, Zhang B, Xu J, Huang F, Zhao RN and Chen YJ: Ibandronate promotes osteogenic differentiation of periodontal ligament stem cells by regulating the expression of microRNAs. Biochem Biophys Res Commun. 404:127–132. 2011. View Article : Google Scholar : PubMed/NCBI
17	Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, Young M, Robey PG, Wang CY and Shi S: Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 364:149–155. 2004. View Article : Google Scholar : PubMed/NCBI
18	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
19	Li B, Qu C, Chen C, Liu Y, Akiyama K, Yang R, Chen F, Zhao Y and Shi S: Basic fibroblast growth factor inhibits osteogenic differentiation of stem cells from human exfoliated deciduous teeth through ERK signaling. Oral Dis. 18:285–292. 2012. View Article : Google Scholar : PubMed/NCBI
20	Han L, Yang Y, Yue X, Huang K, Liu X, Pu P, Jiang H, Yan W, Jiang T and Kang C: Inactivation of PI3K/AKT signaling inhibits glioma cell growth through modulation of β-catenin-mediated transcription. Brain Res. 1366:9–17. 2010. View Article : Google Scholar : PubMed/NCBI
21	Jones-Rhoades MW and Bartel DP: Computational identification of plant microRNAs and their targets, including a stress-induced miRNA. Mol Cell. 14:787–799. 2004. View Article : Google Scholar : PubMed/NCBI
22	Shi K, Lu J, Zhao Y, Wang L, Li J, Qi B, Li H and Ma C: MicroRNA-214 suppresses osteogenic differentiation of C2C12 myoblast cells by targeting Osterix. Bone. 55:487–494. 2013. View Article : Google Scholar : PubMed/NCBI
23	Yang L, Ge D, Cao X, Ge Y, Chen H, Wang W and Zhang H: MiR-214 Attenuates osteogenic differentiation of mesenchymal stem cells via targeting FGFR1. Cell Physiol Biochem. 38:809–820. 2016. View Article : Google Scholar : PubMed/NCBI
24	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
25	Su J, Zhang A, Shi Z, Ma F, Pu P, Wang T, Zhang J, Kang C and Zhang Q: MicroRNA-200a suppresses the Wnt/β-catenin signaling pathway by interacting with β-catenin. Int J Oncol. 40:1162–1170. 2012.PubMed/NCBI
26	Chen H, Mo D, Li M, Zhang Y, Chen L, Zhang X, Li M, Zhou X and Chen Y: miR-709 inhibits 3T3-L1 cell differentiation by targeting GSK3β of Wnt/β-catenin signaling. Cell Signal. 26:2583–2589. 2014. View Article : Google Scholar : PubMed/NCBI
27	Wang T and Xu Z: miR-27 promotes osteoblast differentiation by modulating Wnt signaling. Biochem Biophys Res Commun. 402:186–189. 2010. View Article : Google Scholar : PubMed/NCBI