Abstract
Adipose-derived stem cells (ADSCs) provide a novel method for bone tissue regeneration, but their adipogenic tendency limits their therapeutic efficacy. MicroRNAs (miRNAs) have been reported to regulate stem cell differentiation and bone tissue regeneration, but the detailed mechanism is poorly investigated. Our study indicated that inhibition of miR-145-5p enhanced the osteogenic potential of ADSCs and reduced the adipogenic differentiation. Osteogenesis- and adipogenesis-associated genes were detected by qRT-PCR indicating a corresponding result. Moreover, semaphorin 3A (sema3A) was found to be a target of miR-145-5p, as confirmed by a luciferase activity assay, qRT-PCR, and western blotting. Inhibition of miR-145-5p promoted migration, as detected by wound healing and Transwell assays, but did not affect proliferation, as detected by CCK-8 and ki-67 assays. The effects of miR-145-5p inhibitors on ADSC progression rescued by siRNA of Sema3a and si-sema3a exerted the same effect as miR-145-5p inhibitors on ADSC progression. Furthermore, siRNA of Sema3a rescued synergistic effects with miR-145-5p inhibitors in ADSCs. qRT-PCR and immunofluorescence assays showed that miR-145-5p activated the Wnt signaling pathway for osteogenic differentiation. In conclusion, miR-145-5p and sema3a represent new targets for improving the osteogenic capacity of ADSCs.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297
Behar O, Golden JA, Mashimo H, Schoen FJ, Fishman MC (1996) Semaphorin III is needed for normal patterning and growth of nerves, bones and heart. Nature 383(6600):525–528. https://doi.org/10.1038/383525a0
Boufraqech, M., Zhang, L., Jain, M., Patel, D., Ellis, R., Xiong, Y., . . . Kebebew, E. (2014). miR-145 suppresses thyroid cancer growth and metastasis and targets AKT3. Endocr Relat Cancer, 21(4), 517–531. doi:https://doi.org/10.1530/ERC-14-0077
Bressan, E., Botticelli, D., Sivolella, S., Bengazi, F., Guazzo, R., Sbricoli, L., . . . Zavan, B. (2015). Adipose-derived stem cells as a tool for dental implant osseointegration: an experimental study in the dog. Int J Mol Cell Med, 4(4), 197–208
Bunnell BA, Flaat M, Gagliardi C, Patel B, Ripoll C (2008) Adipose-derived stem cells: isolation, expansion and differentiation. Methods 45(2):115–120
Caplan AI (2009) Why are MSCs therapeutic? New data: new insight. J Pathol 217(2):318–324
Chen D, Li Y, Zhou Z, Xing Y, Yu Z, Zou X et al (2012) Synergistic inhibition of Wnt pathway by HIF-1α and osteoblast-specific transcription factor osterix (Osx) in osteoblasts. PLoS One 7(12):e52948
Fang, K., Song, W., Wang, L., Xu, X., Tan, N., Zhang, S., . . . Song, Y. (2016). Semaphorin 3A-modified adipose-derived stem cell sheet may improve osseointegration in a type 2 diabetes mellitus rat model. Mol Med Rep, 14(3), 2449–2456. doi:https://doi.org/10.3892/mmr.2016.5568
Fukuda, T., Takeda, S., Xu, R., Ochi, H., Sunamura, S., Sato, T., . . . Itoh, H. (2013). Sema3A regulates bone-mass accrual through sensory innervations. Nature, 497(7450), 490–493. doi:https://doi.org/10.1038/nature12115
Gimble JM, Katz AJ, Bunnell BA (2007) Adipose-derived stem cells for regenerative medicine. Circ Res 100(9):1249–1260. https://doi.org/10.1161/01.RES.0000265074.83288.09
Guerrero F, Herencia C, Almadén Y, Martínez-Moreno JM, Montes d OA, Rodriguez-Ortiz ME et al (2014) TGF-β prevents phosphate-induced osteogenesis through inhibition of BMP and Wnt/β-catenin pathways. PLoS One 9(2):e89179
Hao W, Liu H, Zhou L, Sun Y, Su H, Ni J et al (2018) MiR-145 regulates osteogenic differentiation of human adipose-derived mesenchymal stem cells through targeting FoxO1. Exp Biol Med (Maywood) 243(4):386–393. https://doi.org/10.1177/1535370217746611
Hayashi M, Nakashima T, Taniguchi M, Kodama T, Kumanogoh A, Takayanagi H (2012) Osteoprotection by semaphorin 3A. Nature 485(7396):69–74. https://doi.org/10.1038/nature11000
Hkh H, Jiang S, Idris NM, Ashraf M (2008) IGF-1-overexpressing mesenchymal stem cells accelerate bone marrow stem cell mobilization via paracrine activation of SDF-1alpha/CXCR4 signaling to promote myocardial repair. Circ Res 103(11):1300–1308
Hodges WM, O'Brien F, Fulzele S, Hamrick MW (2017) Function of microRNAs in the osteogenic differentiation and therapeutic application of adipose-derived stem cells (ADSCs). Int J Mol Sci 18(12). https://doi.org/10.3390/ijms18122597
Jia J, Tian Q, Ling S, Liu Y, Yang S, Shao Z (2013) miR-145 suppresses osteogenic differentiation by targeting Sp7. FEBS Lett 587(18):3027–3031
Kang, B. J., Ryu, H. H., Park, S. S., Koyama, Y., Kikuchi, M., Woo, H. M., . . . Kweon, O. K. (2012). Comparing the osteogenic potential of canine mesenchymal stem cells derived from adipose tissues, bone marrow, umbilical cord blood, and Wharton's jelly for treating bone defects. J Vet Sci, 13(3), 299–310
Kawasaki T, Bekku Y, Suto F, Kitsukawa T, Taniguchi M, Nagatsu I et al (2002) Requirement of neuropilin 1-mediated Sema3A signals in patterning of the sympathetic nervous system. Development 129(3):671–680
Kou B, Gao Y, Du C, Shi Q, Xu S, Wang CQ et al (2014) miR-145 inhibits invasion of bladder cancer cells by targeting PAK1. Urol Oncol 32(6):846–854. https://doi.org/10.1016/j.urolonc.2014.01.003
Larsson L, Decker AM, Nibali L, Pilipchuk SP, Berglundh T, Giannobile WV (2016) Regenerative medicine for periodontal and peri-implant diseases. J Dent Res 95(3):255–266. https://doi.org/10.1177/0022034515618887
Li KC, Chang YH, Hsu MN, Lo SC, Li WH, Hu YC (2017) Baculovirus-mediated miR-214 knockdown shifts osteoporotic ADSCs differentiation and improves osteoporotic bone defects repair. Sci Rep 7(1):16225. https://doi.org/10.1038/s41598-017-16547-3
Liu X, Tan N, Zhou Y, Zhou X, Chen H, Wei H et al (2016) Semaphorin 3A shifts adipose mesenchymal stem cells towards osteogenic phenotype and promotes bone regeneration in vivo. Stem Cells Int 2016:2545214–2545213. https://doi.org/10.1155/2016/2545214
Long F (2012) Building strong bones: molecular regulation of the osteoblast lineage. Nat Rev Mol Cell Biol 13(1):27–38
Maden CH, Gomes J, Schwarz Q, Davidson K, Tinker A, Ruhrberg C (2012) NRP1 and NRP2 cooperate to regulate gangliogenesis, axon guidance and target innervation in the sympathetic nervous system. Dev Biol 369(2):277–285. https://doi.org/10.1016/j.ydbio.2012.06.026
Majumdar MK, Thiede MA, Haynesworth SE, Bruder SP, Gerson SL (2000) Human marrow-derived mesenchymal stem cells (MSCs) express hematopoietic cytokines and support long-term hematopoiesis when differentiated toward stromal and osteogenic lineages. J Hematother Stem Cell Res 9(6):841–848
Medrano S, Sequeira-Lopez ML, Gomez RA (2014) Deletion of the miR-143/145 cluster leads to hydronephrosis in mice. Am J Pathol 184(12):3226–3238. https://doi.org/10.1016/j.ajpath.2014.08.012
Pasquinelli AE, Hunter S, Bracht J (2005) MicroRNAs: a developing story. Curr Opin Genet Dev 15(2):200–205. https://doi.org/10.1016/j.gde.2005.01.002
Qiao Q, Xu X, Song Y, Song S, Zhu W, Li F (2018) Semaphorin 3A promotes osteogenic differentiation of BMSC from type 2 diabetes mellitus rats. J Mol Histol 49:369–376. https://doi.org/10.1007/s10735-018-9776-1
Schipper BM, Marra KG, Zhang W, Donnenberg AD, Rubin JP (2008) Regional anatomic and age effects on cell function of human adipose-derived stem cells. Ann Plast Surg 60(5):538–544
Shen H, Shen J, Wang L, Shi Z, Wang M, Jiang BH, Shu Y (2015) Low miR-145 expression level is associated with poor pathological differentiation and poor prognosis in non-small cell lung cancer. Biomed Pharmacother 69:301–305. https://doi.org/10.1016/j.biopha.2014.12.019
Shen S, Yaqian LU, Zheng Y, Zhang W, & Zhu Q (2017) The study of microRNA-145(miR-145) modulating osteogenesis differentiation of rat bone marrow mesenchymal stem cells (rBMMSC). Stomatology
Toyofuku T, Yoshida J, Sugimoto T, Zhang H, Kumanogoh A, Hori M, Kikutani H (2005) FARP2 triggers signals for Sema3A-mediated axonal repulsion. Nat Neurosci 8(12):1712–1719. https://doi.org/10.1038/nn1596
Yokoyama N, Yin D, Malbon C (2007) Abundance, complexation, and trafficking of Wnt/β-catenin signaling elements in response to Wnt3a. J Mol Signal 2(1):1–16
Zaminy A, Ragerdi KI, Barbarestani M, Hedayatpour A, Mahmoudi R, Farzaneh NA (2008) Osteogenic differentiation of rat mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells: melatonin as a differentiation factor. Iran Biomed J 12(3):133
Zhao H, Kang X, Xia X, Wo L, Gu X, Hu Y et al (2016) miR-145 suppresses breast cancer cell migration by targeting FSCN-1 and inhibiting epithelial-mesenchymal transition. Am J Transl Res 8(7):3106–3114
Zuk PA (2010) The adipose-derived stem cell: looking back and looking ahead. Mol Biol Cell 21(11):1783–1787. https://doi.org/10.1091/mbc.E09-07-0589
Funding
This study was funded by the Natural Science Foundation of China (NSFC) Nos. 81470775 and 81771107.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Editor: Tetsuji Okamoto
Electronic supplementary material
ESM 1
(DOCX 283 kb)
Rights and permissions
About this article
Cite this article
Liu, X., Zhu, W., Wang, L. et al. miR-145-5p suppresses osteogenic differentiation of adipose-derived stem cells by targeting semaphorin 3A. In Vitro Cell.Dev.Biol.-Animal 55, 189–202 (2019). https://doi.org/10.1007/s11626-019-00318-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11626-019-00318-7