miR‑29b promotes the osteogenic differentiation of mesenchymal stem cells derived from human adipose tissue via the PTEN/AKT/β‑catenin signaling pathway

Xia,Tian; Dong,Shuanghai; Tian,Jiwei

doi:10.3892/ijmm.2020.4615

miR‑29b promotes the osteogenic differentiation of mesenchymal stem cells derived from human adipose tissue via the PTEN/AKT/β‑catenin signaling pathway

Authors:
- Tian Xia
- Shuanghai Dong
- Jiwei Tian
View Affiliations

Affiliations: Shanghai General Hospital of Nanjing Medical University, Shanghai 200080, P.R. China, Department of Orthopedics, Shanghai Jiahui International Hospital, Shanghai 200233, P.R. China
Published online on: May 22, 2020 https://doi.org/10.3892/ijmm.2020.4615
Pages: 709-717
Copyright: © Xia 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

Accumulating evidence has documented that microRNAs (miRNAs or miRs) function as important post‑transcriptional regulators of the differentiation of mesenchymal stem cells (MSCs), including human adipose‑derived mesenchymal stem cells (hADSCs); however, their roles in hADSC osteogenic differentiation require further investigation. The present study aimed to investigate the role of miRNAs in the osteogenic differentiation of hADSCs and to elucidate the underlying molecular mechanisms. Using an miRNA microarray, it was found that 24 miRNAs were upregulated and 14 miRNAs were downregulated compared with the undifferentiated cells, and miR‑29b‑3p (miR‑29b) was selected for further experiments. Functional experiments revealed that the upregulation of miR‑29b by agomir‑29b significantly enhanced alkaline phosphatase (ALP) activity and the mineralization of extracellular matrix (ECM), and led to an increase in the mRNA and protein levels of osteogenic marker genes, including runt‑related transcription factor 2 (Runx2), osteopontin (OPN), osteocalcin (OCN) and bone sialoprotein (BSP), whereas the knockdown of miR‑29b suppressed these processes. In addition, phosphatase and tensin homolog (PTEN), a negative regulator of the AKT/β‑catenin pathway, was identified as a direct target of miR‑29b in the hADSCs. Moreover, it was observed that the overexpression of miR‑29b activated the AKT/β‑catenin signaling pathway by inhibiting PTEN expression in the hADSCs. Most importantly, it was also found that the overexpression of PTEN reversed the promoting effects of miR‑29b on osteogenic differentiation. On the whole, these findings suggest that miR‑29b promotes the osteogenic differentiation of hADSCs by modulating the PTEN/AKT/β‑catenin signaling pathway. Thus, this miRNA may be a promising target for the active modulation of hADSC‑derived osteogenesis.

View Figures

View References

1	Deng Y, Zhou H, Zou D, Xie Q, Bi X, Gu P and Fan X: The role of miR-31-modified adipose tissue-derived stem cells in repairing rat critical-sized calvarial defects. Biomaterials. 34:6717–6728. 2013. View Article : Google Scholar : PubMed/NCBI
2	Lin CY, Chang YH, Li KC, Lu CH, Sung LY, Yeh CL, Lin KJ, Huang SF, Yen TC and Hu YC: The use of ASCs engineered to express BMP2 or TGF-β3 within scaffold constructs to promote calvarial bone repair. Biomaterials. 34:9401–9412. 2013. View Article : Google Scholar : PubMed/NCBI
3	Locke M, Feisst V and Dunbar PR: Concise review: Human adipose-derived stem cells: Separating promise from clinical need. Stem Cells. 29:404–411. 2011. View Article : Google Scholar : PubMed/NCBI
4	Gadkari R, Zhao L, Teklemariam T and Hantash BM: Human embryonic stem cell derived-mesenchymal stem cells: An alternative mesenchymal stem cell source for regenerative medicine therapy. Regen Med. 9:453–465. 2014. View Article : Google Scholar : PubMed/NCBI
5	Song F, Jiang D, Wang T, Wang Y, Lou Y, Zhang Y, Ma H and Kang Y: Mechanical stress regulates osteogenesis and adipogenesis of rat mesenchymal stem cells through PI3K/Akt/GSK-3β/β-catenin signaling pathway. Biomed Res Int. 2017:60274022017. View Article : Google Scholar
6	Wu X, Zheng S, Ye Y, Wu Y, Lin K and Su J: Enhanced osteogenic differentiation and bone regeneration of poly(lactic-co-glycolic acid) by graphene via activation of PI3K/Akt/GSK-3β/β-catenin signal circuit. Biomater Sci. 6:1147–1158. 2018. View Article : Google Scholar : PubMed/NCBI
7	Saidak Z, Le Henaff C, Azzi S, Marty C, Da Nascimento S, Sonnet P and Marie PJ: Wnt/β-catenin signaling mediates osteoblast differentiation triggered by peptide-induced α5β1 integrin priming in mesenchymal skeletal cells. J Biol Chem. 290:6903–6912. 2015. View Article : Google Scholar : PubMed/NCBI
8	Chen JC and Jacobs CR: Mechanically induced osteogenic lineage commitment of stem cells. Stem Cell Res Ther. 4:1072013. View Article : Google Scholar : PubMed/NCBI
9	Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI
10	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
11	Luzi E, Marini F, Sala SC, Tognarini I, Galli G and Brandi ML: Osteogenic differentiation of human adipose tissue-derived stem cells is modulated by the miR-26a targeting of the SMAD1 transcription factor. J Bone Miner Res. 23:287–295. 2008. View Article : Google Scholar : PubMed/NCBI
12	Li J, Hu C, Han L, Liu L, Jing W, Tang W, Tian W and Long J: MiR-154-5p regulates osteogenic differentiation of adipose-derived mesenchymal stem cells under tensile stress through the Wnt/PCP pathway by targeting Wnt11. Bone. 78:130–141. 2015. View Article : Google Scholar : PubMed/NCBI
13	Kim YJ, Bae SW, Yu SS, Bae YC and Jung JS: miR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue. J Bone Miner Res. 24:816–825. 2009. View Article : Google Scholar
14	Chen S, Zheng Y, Zhang S, Jia L and Zhou Y: Promotion effects of miR-375 on the osteogenic differentiation of human adipose-derived mesenchymal stem cells. Stem Cell Reports. 8:773–786. 2017. View Article : Google Scholar : PubMed/NCBI
15	Peng S, Gao D, Gao C, Wei P, Niu M and Shuai C: MicroRNAs regulate signaling pathways in osteogenic differentiation of mesenchymal stem cells (Review). Mol Med Rep. 14:623–629. 2016. View Article : Google Scholar : PubMed/NCBI
16	Boquest AC, Shahdadfar A, Brinchmann JE and Collas P: Isolation of stromal stem cells from human adipose tissue. Methods Mol Biol. 325:35–46. 2006.PubMed/NCBI
17	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
18	Mei LL, Wang WJ, Qiu YT, Xie XF, Bai J and Shi ZZ: miR-125b-5p functions as a tumor suppressor gene partially by regulating HMGA2 in esophageal squamous cell carcinoma. PLoS One. 12:e01856362017. View Article : Google Scholar : PubMed/NCBI
19	Zhang MY, Lin J and Kui YC: MicroRNA-345 suppresses cell invasion and migration in non-small cell lung cancer by directly targeting YAP1. Eur Rev Med Pharmacol Sci. 23:2436–2443. 2019.PubMed/NCBI
20	Fang S, Deng Y, Gu P and Fan X: MicroRNAs regulate bone development and regeneration. Int J Mol Sci. 16:8227–8253. 2015. View Article : Google Scholar : PubMed/NCBI
21	Tang P, Xiong Q, Ge W and Zhang L: The role of microRNAs in osteoclasts and osteoporosis. RNA Biol. 11:1355–1363. 2014. View Article : Google Scholar
22	Roberto VP, Tiago DM, Silva IA and Cancela ML: MiR-29a is an enhancer of mineral deposition in bone-derived systems. Arch Biochem Biophys. 564:173–183. 2014. View Article : Google Scholar : PubMed/NCBI
23	Li Z, Hassan MQ, Jafferji M, Aqeilan RI, Garzon R, Croce CM, van Wijnen AJ, Stein JL, Stein GS and Lian JB: Biological functions of miR-29b contribute to positive regulation of osteoblast differentiation. J Biol Chem. 284:15676–15684. 2009. View Article : Google Scholar : PubMed/NCBI
24	Mayer U, Benditz A and Grässel S: miR-29b regulates expression of collagens I and III in chondrogenically differentiating BMSC in an osteoarthritic environment. Sci Rep. 7:132972017. View Article : Google Scholar : PubMed/NCBI
25	Wang W and Cao W: Treatment of osteoarthritis with mesenchymal stem cells. Sci China Life Sci. 57:586–595. 2014. View Article : Google Scholar : PubMed/NCBI
26	Baharlou R, Ahmadi-Vasmehjani A, Faraji F, Atashzar MR, Khoubyari M, Ahi S, Erfanian S and Navabi SS: Human adipose tissue-derived mesenchymal stem cells in rheumatoid arthritis: Regulatory effects on peripheral blood mononuclear cells activation. Int Immunopharmacol. 47:59–69. 2017. View Article : Google Scholar : PubMed/NCBI
27	Meng YB, Li X, Li ZY, Zhao J, Yuan XB, Ren Y, Cui ZD, Liu YD and Yang XJ: microRNA-21-promotes osteogenic differentiation of mesenchymal stem cells by the PI3K/β-catenin pathway. J Orthop Res. 33:957–964. 2015. View Article : Google Scholar : PubMed/NCBI
28	Shen GY, Ren H, Huang JJ, Zhang ZD, Zhao WH, Yu X, Shang Q, Qiu T, Zhang YZ, Tang JJ, et al: Plastrum testudinis extracts promote BMSC proliferation and osteogenic differentiation by regulating let-7f-5p and the TNFR2/PI3K/AKT signaling pathway. Cell Physiol Biochem. 47:2307–2318. 2018. View Article : Google Scholar : PubMed/NCBI
29	Xie Q, Wang Z, Zhou H, Yu Z, Huang Y, Sun H, Bi X, Wang Y, Shi W, Gu P and Fan X: The role of miR-135-modified adipose-derived mesenchymal stem cells in bone regeneration. Biomaterials. 75:279–294. 2016. View Article : Google Scholar
30	Hu R, Li H, Liu W, Yang L, Tan YF and Luo XH: Targeting miRNAs in osteoblast differentiation and bone formation. Expert Opin Ther Targets. 14:1109–1120. 2010. View Article : Google Scholar : PubMed/NCBI
31	Cho HH, Shin KK, Kim YJ, Song JS, Kim JM, Bae YC, Kim CD and Jung JS: NF-kappaB activation stimulates osteogenic differentiation of mesenchymal stem cells derived from human adipose tissue by increasing TAZ expression. J Cell Physiol. 223:168–177. 2010.PubMed/NCBI
32	Huang S, Wang S, Bian C, Yang Z, Zhou H, Zeng Y, Li H, Han Q and Zhao RC: Upregulation of miR-22-promotes osteogenic differentiation and inhibits adipogenic differentiation of human adipose tissue-derived mesenchymal stem cells by repressing HDAC 6-protein expression. Stem Cells Dev. 21:2531–2540. 2012. View Article : Google Scholar : PubMed/NCBI
33	Zeng Y, Qu X, Li H, Huang S, Wang S, Xu Q, Lin R, Han Q, Li J and Zhao RC: MicroRNA-100 regulates osteogenic differentiation of human adipose-derived mesenchymal stem cells by targeting BMPR2. FEBS Lett. 586:2375–2381. 2012. View Article : Google Scholar : PubMed/NCBI
34	Wei J, Shi Y, Zheng L, Zhou B, Inose H, Wang J, Guo XE, Grosschedl R and Karsenty G: miR-34s inhibit osteoblast proliferation and differentiation in the mouse by targeting SATB2. J Cell Biol. 197:509–521. 2012. View Article : Google Scholar : PubMed/NCBI
35	Nielsen-Preiss SM, Silva SR and Gillette JM: Role of PTEN and Akt in the regulation of growth and apoptosis in human osteoblastic cells. J Cell Biochem. 90:964–975. 2003. View Article : Google Scholar : PubMed/NCBI
36	Zhang Y, Zhang L, Yan M and Zheng X: Inhibition of phosphatidylinositol 3-kinase causes cell death in rat osteoblasts through inactivation of Akt. Biomed Pharmacother. 61:277–284. 2007. View Article : Google Scholar : PubMed/NCBI
37	Kawamura N, Kugimiya F, Oshima Y, Ohba S, Ikeda T, Saito T, Shinoda Y, Kawasaki Y, Ogata N, Hoshi K, et al: Akt1 in osteoblasts and osteoclasts controls bone remodeling. PLoS One. 2:e10582007. View Article : Google Scholar : PubMed/NCBI
38	Shen Y, Zhang J, Yu T and Qi C: Generation of PTEN knockout bone marrow mesenchymal stem cell lines by CRISPR/Cas9-mediated genome editing. Cytotechnology. 70:783–791. 2018. View Article : Google Scholar : PubMed/NCBI
39	Liu X, Chen T, Wu Y and Tang Z: Role and mechanism of PTEN in adiponectin-induced osteogenesis in human bone marrow mesenchymal stem cells. Biochem Biophys Res Commun. 483:712–717. 2017. View Article : Google Scholar
40	Lin X, Zhou X, Liu D, Yun L, Zhang L, Chen X, Chai Q and Li L: MicroRNA-29 regulates high-glucose-induced apoptosis in human retinal pigment epithelial cells through PTEN. In. Vitro Cell Dev Biol Anim. 52:419–426. 2016. View Article : Google Scholar
41	Sen B, Styner M, Xie Z, Case N, Rubin CT and Rubin J: Mechanical loading regulates NFATc1 and beta-catenin signaling through a GSK3beta control node. J Biol Chem. 284:34607–34617. 2009. View Article : Google Scholar : PubMed/NCBI
42	Day TF, Guo X, Garrett-Beal L and Yang Y: Wnt/beta-catenin signaling in mesenchymal progenitors controls osteoblast and chondrocyte differentiation during vertebrate skeletogenesis. Dev Cell. 8:739–750. 2005. View Article : Google Scholar : PubMed/NCBI
43	Qu ZH, Zhang XL, Tang TT and Dai KR: Promotion of osteogenesis through beta-catenin signaling by desferrioxamine. Biochem Biophys Res Commun. 370:332–337. 2008. View Article : Google Scholar : PubMed/NCBI
44	Steck PA, Pershouse MA, Jasser SA, Yung WK, Lin H, Ligon AH, Langford LA, Baumgard ML, Hattier T, Davis T, et al: Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers. Nat Genet. 15:356–362. 1997. View Article : Google Scholar : PubMed/NCBI
45	Whang YE, Wu X, Suzuki H, Reiter RE, Tran C, Vessella RL, Said JW, Isaacs WB and Sawyers CL: Inactivation of the tumor suppressor PTEN/MMAC1 in advanced human prostate cancer through loss of expression. Proc Natl Acad Sci USA. 95:5246–5250. 1998. View Article : Google Scholar : PubMed/NCBI