International Journal of Stem Cells

  • 제15권3호
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  • Pages.324-333
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  • 2022
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  • 2005-3606(pISSN)
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  • 2005-5447(eISSN)
한국줄기세포학회 (Korean Society for Stem Cell Research)
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MiR-29a-3p Inhibits Proliferation and Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells via Targeting FOXO3 and Repressing Wnt/β-Catenin Signaling in Steroid-Associated Osteonecrosis

  • Changgeng Wang (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang) ;
  • Minghui Zhu (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang) ;
  • Demeng Yang (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang) ;
  • Xinyuan Hu (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang) ;
  • Xinyuan Wen (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang) ;
  • Aimei Liu (Department of Emergency and Trauma Surgery, People's Hospital of Pingxiang)
  • 투고 : 2021.08.29
  • 심사 : 2022.03.30
  • 발행 : 2022.08.30
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초록

Background and Objectives: This study was to investigate the role of microRNA-29a-3p (miR-29a-3p) in human bone marrow mesenchymal stem cells (hBMSCs), and its relationship with steroid-associated osteonecrosis. Methods and Results: The online tool GEO2R was used to screen out the differentially expressed genes (DEGs) in GSE123568 dataset. Quantitative real time-polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-29a-3p, forkhead box O3 (FOXO3), alkaline phosphatase (ALP), bone gamma-carboxyglutamate protein (OCN) and RUNX family transcription factor 2 (Runx2) in the hBMSCs isolated from the patients with steroid-associated osteonecrosis. CCK-8 assay was executed to measure cell viability; western blot assay was utilized to detect FOXO3, ALP, Runx2, OCN and β-catenin expression. Cell apoptosis and cell cycle were detected by flow cytometry. Immunofluorescence assay was used to detect the sub-cellular localization of β-catenin. Bioinformatics analysis and luciferase reporter gene assay were performed to confirm whether miR-29a-3p can combine with FOXO3 3'UTR. MiR-29a-3p was markedly up-regulated in the hBMSCs of patients with steroid-associated osteonecrosis, while FOXO3 mRNA was significantly down-regulated. Transfection of miR-29a-3p mimics significantly inhibited the hBMSCs' proliferation, osteogenic differentiation markers' expressions, including ALP, Runx2, OCN, and repressed the ALP activity, as well as promoted cell apoptosis and cell-cycle arrest. FOXO3 was identified as a target gene of miR-29a-3p, and miR-29a-3p can inhibit the expression of FOXO3 and β-catenin, and inhibition of miR-29a-3p promoted translocation of β-catenin to the nucleus. Conclusions: MiR-29a-3p can modulate FOXO3 expression and Wnt/β-catenin signaling to inhibit viability and osteogenic differentiation of hBMSCs, thereby promoting the development of steroid-associated osteonecrosis.

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참고문헌

  1. Bose VC, Baruah BD. Resurfacing arthroplasty of the hip for avascular necrosis of the femoral head: a minimum follow-up of four years. J Bone Joint Surg Br 2010;92:922-928
  2. Yue J, Wan F, Zhang Q, Wen P, Cheng L, Li P, Guo W. Effect of glucocorticoids on miRNA expression spectrum of rat femoral head microcirculation endothelial cells. Gene 2018;651:126-133
  3. Su P, Tian Y, Yang C, Ma X, Wang X, Pei J, Qian A. Mesenchymal stem cell migration during bone formation and bone diseases therapy. Int J Mol Sci 2018;19:2343
  4. Park KR, Kim S, Cho M, Kang SW, Yun HM. Effects of PIN on osteoblast differentiation and matrix mineralization through Runt-related transcription factor. Int J Mol Sci 2020;21:9579
  5. Deng S, Dai G, Chen S, Nie Z, Zhou J, Fang H, Peng H. Dexamethasone induces osteoblast apoptosis through ROS-PI3K/AKT/GSK3β signaling pathway. Biomed Pharmacother 2019;110:602-608
  6. Beyer C, Zampetaki A, Lin NY, Kleyer A, Perricone C, Iagnocco A, Distler A, Langley SR, Gelse K, Sesselmann S, Lorenzini R, Niemeier A, Swoboda B, Distler JH, Santer P, Egger G, Willeit J, Mayr M, Schett G, Kiechl S. Signature of circulating microRNAs in osteoarthritis. Ann Rheum Dis 2015;74:e18
  7. Wang J, Liu S, Li J, Zhao S, Yi Z. Roles for miRNAs in osteogenic differentiation of bone marrow mesenchymal stem cells. Stem Cell Res Ther 2019;10:197
  8. Liu Y, Zong Y, Shan H, Lin Y, Xia W, Wang N, Zhou L, Gao Y, Ma X, Jiang C. MicroRNA-23b-3p participates in steroid-induced osteonecrosis of the femoral head by suppressing ZNF667 expression. Steroids 2020;163:108709
  9. Zhao JJ, Wu ZF, Wang L, Feng DH, Cheng L. MicroRNA145 mediates steroid-induced necrosis of the femoral head by targeting the OPG/RANK/RANKL signaling pathway. PLoS One 2016;11:e0159805
  10. Dai Z, Jin Y, Zheng J, Liu K, Zhao J, Zhang S, Wu F, Sun Z. MiR-217 promotes cell proliferation and osteogenic differentiation of BMSCs by targeting DKK1 in steroid-associated osteonecrosis. Biomed Pharmacother 2019;109:1112-1119
  11. Jin H, Wang H, Jin X, Wang W. Long non-coding RNA H19 regulates LASP1 expression in osteosarcoma by competitively binding to miR-29a-3p. Oncol Rep 2021;46:207
  12. Muluhngwi P, Klinge CM. Identification and roles of miR-29b-1-3p and miR29a-3p-regulated and non-regulated lncRNAs in endocrine-sensitive and resistant breast cancer cells. Cancers (Basel) 2021;13:3530
  13. Geng A, Luo L, Ren F, Zhang L, Zhou H, Gao X. miR-29a-3p inhibits endometrial cancer cell proliferation, migration and invasion by targeting VEGFA/CD C42/PAK1. BMC Cancer 2021;21:843
  14. Zhou W, Li H, Shang S, Liu F. lncRNA KCNQ1OT1 reverses the effect of sevoflurane on hepatocellular carcinoma progression via regulating the miR-29a-3p/CBX3 axis. Braz J Med Biol Res 2021;54:e10213
  15. Salih DA, Brunet A. FoxO transcription factors in the maintenance of cellular homeostasis during aging. Curr Opin Cell Biol 2008;20:126-136
  16. Gomez-Puerto MC, Verhagen LP, Braat AK, Lam EW, Coffer PJ, Lorenowicz MJ. Activation of autophagy by FOXO3 regulates redox homeostasis during osteogenic differentiation. Autophagy 2016;12:1804-1816
  17. Ambrogini E, Almeida M, Martin-Millan M, Paik JH, Depinho RA, Han L, Goellner J, Weinstein RS, Jilka RL, O'Brien CA, Manolagas SC. FoxO-mediated defense against oxidative stress in osteoblasts is indispensable for skeletal homeostasis in mice. Cell Metab 2010;11:136-146
  18. Han S, Wang Z, Liu J, Wang HD, Yuan Q. miR-29a-3p-dependent COL3A1 and COL5A1 expression reduction assists sulforaphane to inhibit gastric cancer progression. Biochem Pharmacol 2021;188:114539
  19. Liu F, Wang Z, Liu F, Xu J, Liu Q, Yin K, Lan J. MicroRNA-29a-3p enhances dental implant osseointegration of hyperlipidemic rats via suppressing dishevelled 2 and frizzled 4. Cell Biosci 2018;8:55
  20. Zeng X, Wang Y, Dong Q, Ma MX, Liu XD. DLX2 activates Wnt1 transcription and mediates Wnt/β-catenin signal to promote osteogenic differentiation of hBMSCs. Gene 2020;744:144564
  21. Wu X, Sun W, Tan M. Noncoding RNAs in steroid-induced osteonecrosis of the femoral head. Biomed Res Int 2019;2019:8140595
  22. Chen Y, Zhang W, Yan L, Zheng P, Li J. miR-29a-3p directly targets Smad nuclear interacting protein 1 and inhibits the migration and proliferation of cervical cancer HeLa cells. PeerJ 2020;8:e10148
  23. Zhu D, Tian J, Wu X, Li M, Tang X, Rui K, Guo H, Ma J, Xu H, Wang S. G-MDSC-derived exosomes attenuate collagen-induced arthritis by impairing Th1 and Th17 cell responses. Biochim Biophys Acta Mol Basis Dis 2019;1865:165540
  24. Chen D, Gong Y, Xu L, Zhou M, Li J, Song J. Bidirectional regulation of osteogenic differentiation by the FOXO subfamily of Forkhead transcription factors in mammalian MSCs. Cell Prolif 2019;52:e12540
  25. Long C, Cen S, Zhong Z, Zhou C, Zhong G. FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy. Hum Cell 2021;34:14-27
  26. Guerit D, Brondello JM, Chuchana P, Philipot D, Toupet K, Bony C, Jorgensen C, Noel D. FOXO3A regulation by miRNA-29a controls chondrogenic differentiation of mesenchymal stem cells and cartilage formation. Stem Cells Dev 2014;23:1195-1205
  27. Fan J, An X, Yang Y, Xu H, Fan L, Deng L, Li T, Weng X, Zhang J, Chunhua Zhao R. MiR-1292 targets FZD4 to regulate senescence and osteogenic differentiation of stem cells in TE/SJ/mesenchymal tissue system via the Wnt/β-catenin pathway. Aging Dis 20184;9:1103-1121
  28. Tu X, Delgado-Calle J, Condon KW, Maycas M, Zhang H, Carlesso N, Taketo MM, Burr DB, Plotkin LI, Bellido T. Osteocytes mediate the anabolic actions of canonical Wnt/β-catenin signaling in bone. Proc Natl Acad Sci U S A 2015;112:E478-E486
  29. Pei J, Fan L, Nan K, Li J, Shi Z, Dang X, Wang K. Excessive activation of TLR4/NF-κB interactively suppresses the canonical Wnt/β-catenin pathway and induces SANFH in SD rats. Sci Rep 2017;7:11928