MicroRNA expression profiles in human adipose-derived stem cells during chondrogenic differentiation

Yang,Zhen; Hao,Jie; Hu,Zhen‑Ming

doi:10.3892/ijmm.2014.2051

MicroRNA expression profiles in human adipose-derived stem cells during chondrogenic differentiation

Authors:
- Zhen Yang
- Jie Hao
- Zhen‑Ming Hu
View Affiliations

Affiliations: Department of Orthopedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: December 29, 2014 https://doi.org/10.3892/ijmm.2014.2051
Pages: 579-586
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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 aim of the present study was to examine the microRNA (miRNA or miR) expression profiles during the chondrogenic differentiation of human adipose‑derived stem cells (hADSCs) and identify the potential mechanisms through which miRNAs may affect the process of chondrogenesis. hADSCs were isolated and cultured. The expression levels of chondrogenic markers was detected by FACS analysis and immunohistochemistry. The miRNA expression profiles were then obtained through a miRNA array and confirmed through northern blot analysis. Putative targets of the miRNAs were predicted and validated through a luciferase reporter assay. The comparison of hADSCs following the induction of chondrogenic differentiation with undifferentiated hADSCs revealed 20 miRNAs that were differentially expressed by at least 2‑fold, and these miRNAs included 12 upregulated miRNAs and 8 downregulated miRNAs. Northern blot analysis further conﬁrmed the miRNA expression levels. Of these miRNAs, the expression of miR‑490‑5p was gradually downregulated following the induction of chondrogenic differentiation. The overexpression of miR‑490‑5p increased the expression of the chondrogenic markers, collagen, type II, alpha 1 (Col2A1), collagen, type X, alpha 1 (Col10A1) and aggrecan. Furthermore, it was confirmed that miR‑490‑5p directly targets bone morphogenetic protein receptor type 2 (BMPR2). In conclusion, in this study, we identified a set of miRNAs that may play key roles in the regulation of the chondrogenic differentiation of hADSCs. Our results may provide a basis for the further investigations into the molecular mechanisms of action of miRNAs in hADSC chondrogenesis.

View Figures

View References

1	Dong S, Yang B, Guo H and Kang F: MicroRNAs regulate osteogenesis and chondrogenesis. Biochem Biophys Res Commun. 418:587–591. 2012. View Article : Google Scholar : PubMed/NCBI
2	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.
3	Tomé M, López-Romero P, Albo C, Sepúlveda JC, Fernández-Gutiérrez B, Dopazo A, Bernad A and González MA: miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells. Cell Death Differ. 18:985–995. 2011. View Article : Google Scholar :
4	Wagner W, Horn P, Castoldi M, Diehlmann A, Bork S, Saffrich R, Benes V, Blake J, Pfister S, Eckstein V and Ho AD: Replicative senescence of mesenchymal stem cells: a continuous and organized process. PloS One. 3:e22132008. View Article : Google Scholar : PubMed/NCBI
5	Ning G, Liu X, Dai M, Meng A and Wang Q: MicroRNA-92a upholds Bmp signaling by targeting noggin3 during pharyngeal cartilage formation. Dev Cell. 24:283–295. 2013. View Article : Google Scholar : PubMed/NCBI
6	Song J, Lee M, Kim D, Han J, Chun CH and Jin EJ: MicroRNA-181b regulates articular chondrocytes differentiation and cartilage integrity. Biochem Biophys Res Commun. 431:210–214. 2013. View Article : Google Scholar : PubMed/NCBI
7	Karlsen TA, Jakobsen RB, Mikkelsen TS and Brinchmann JE: microRNA-140 targets RALA and regulates chondrogenic differentiation of human mesenchymal stem cells by translational enhancement of SOX9 and ACAN. Stem Cells Dev. 23:290–304. 2014. View Article : Google Scholar
8	Buechli ME, Lamarre J and Koch TG: MicroRNA-140 expression during chondrogenic differentiation of equine cord blood-derived mesenchymal stromal cells. Stem Cells Dev. 22:1288–1296. 2013. View Article : Google Scholar
9	Ham O, Song BW, Lee SY, Choi E, Cha MJ, Lee CY, Park JH, Kim IK, Chang W, Lim S, Lee CH, Kim S, Jang Y and Hwang KC: The role of microRNA-23b in the differentiation of MSC into chondrocyte by targeting protein kinase A signaling. Biomaterials. 33:4500–4507. 2012. View Article : Google Scholar : PubMed/NCBI
10	Bossio C, Mastrangelo R, Morini R, Tonna N, Coco S, Verderio C, Matteoli M and Bianco F: A simple method to generate adipose stem cell-derived neurons for screening purposes. J Mol Neurosci. 51:274–281. 2013. View Article : Google Scholar : PubMed/NCBI
11	Pandey AC, Semon JA, Kaushal D, O’Sullivan RP, Glowacki J, Gimble JM and Bunnell BA: MicroRNA profiling reveals age-dependent differential expression of nuclear factor κB and mitogen-activated protein kinase in adipose and bone marrow-derived human mesenchymal stem cells. Stem Cell Res Ther. 2(49): 2011
12	Crobu F, Latini V, Marongiu MF, Sogos V, Scintu F, Porcu S, Casu C, Badiali M, Sanna A, Manchinu MF and Ristaldi MS: Differentiation of single cell derived human mesenchymal stem cells into cells with a neuronal phenotype: RNA and microRNA expression profile. Mol Biol Rep. 39:3995–4007. 2012. View Article : Google Scholar
13	Guan L, Shaoqing L, Wang Y, Yue H, Liu D, He L, Bai C, Yan F, Nan X, Shi S and Pei X: In vitro differentiation of human adipose-derived mesenchymal stem cells into endothelial-like cells. Chinese Science Bulletin. 51:1863–1868. 2006. View Article : Google Scholar
14	Zhu Y, Liu T, Song K, Fan X, Ma X and Cui Z: Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochem Funct. 26:664–675. 2008. View Article : Google Scholar : PubMed/NCBI
15	Peroni D, Scambi I, Pasini A, Lisi V, Bifari F, Krampera M, Rigotti G, Sbarbati A and Galiè M: Stem molecular signature of adipose-derived stromal cells. Exp Cell Res. 314:603–615. 2008. View Article : Google Scholar
16	Gronthos S, Franklin DM, Leddy HA, Robey PG, Storms RW and Gimble JM: Surface protein characterization of human adipose tissue-derived stromal cells. J Cell Physiol. 189:54–63. 2001. View Article : Google Scholar : PubMed/NCBI
17	Chen FH, Rousche KT and Tuan RS: Technology Insight: adult stem cells in cartilage regeneration and tissue engineering. Nat Clin Pract Rheumatol. 2:373–382. 2006. View Article : Google Scholar : PubMed/NCBI
18	Vilquin JT and Rosset P: Mesenchymal stem cells in bone and cartilage repair: current status. Regen Med. 1:589–604. 2006. View Article : Google Scholar
19	Iliopoulos D, Malizos KN, Oikonomou P and Tsezou A: Integrative microRNA and proteomic approaches identify novel osteoarthritis genes and their collaborative metabolic and inflammatory networks. PLoS One. 3:e37402008. View Article : Google Scholar : PubMed/NCBI
20	Han J, Yang T, Gao J, Wu J, Qiu X, Fan Q and Ma B: Specific microRNA expression during chondrogenesis of human mesenchymal stem cells. Int J Mol Med. 25:377–384. 2010.PubMed/NCBI
21	Bakhshandeh B, Soleimani M, Paylakhi SH and Ghaemi N: A microRNA signature associated with chondrogenic lineage commitment. J Genet. 91:171–182. 2012. View Article : Google Scholar : PubMed/NCBI
22	Zhang Z, Kang Y, Zhang Z, Zhang H, Duan X, Liu J, Li X and Liao W: Expression of microRNAs during chondrogenesis of human adipose-derived stem cells. Osteoarthritis Cartilage. 20:1638–1646. 2012. View Article : Google Scholar : PubMed/NCBI
23	Indrawattana N, Chen G, Tadokoro M, Shann LH, Ohgushi H, Tateishi T, Tanaka J and Bunyaratvej A: Growth factor combination for chondrogenic induction from human mesenchymal stem cell. Biochem Biophys Res Commun. 320:914–919. 2004. View Article : Google Scholar : PubMed/NCBI
24	Ronzière MC, Perrier E, Mallein-Gerin F and Freyria AM: Chondrogenic potential of bone marrow- and adipose tissue-derived adult human mesenchymal stem cells. Biomed Mater Eng. 20:145–158. 2010.PubMed/NCBI
25	Zhao Q, Eberspaecher H, Lefebvre V and De Crombrugghe B: Parallel expression of Sox9 and Col2a1 in cells undergoing chondrogenesis. Dev Dyn. 209:377–386. 1997. View Article : Google Scholar : PubMed/NCBI