miR-889-3p inhibits osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting Runx2

doi:10.12307/2023.616

Abstract

Abstract: BACKGROUND: Studies have shown that miR-889-3p is able to participate in the regulation of the proliferation and differentiation of a variety of cells, but its effect on the osteogenic differentiation of adipose-derived mesenchymal stem cells remains unclear.
OBJECTIVE: To investigate the regulation of miR-889-3p in osteogenic differentiation of adipose-derived mesenchymal stem cells.
METHODS: Adipose-derived mesenchymal stem cells from SD rats were isolated and cultured in vitro, transfected with miR-889-3p mimics, Mir-889-3p inhibitor and negative control (miR-NC) then induced osteogenic differentiation. RT-PCR and western blot assay were used to detect alkaline phosphatase activity, Runx2, osteocalcin, osteopontin, Osterix and other osteogenic related mRNA and protein expression levels. Adipose-derived mesenchymal stem cells were co-transfected with wild-type Runx2 and mutant Runx2 with miR-889-3p mimic and miR-NC, respectively. Luciferase reporter gene assay was used to detect the targeting relationship between miR-889-3p and Runx2.
RESULTS AND CONCLUSION: (1) The expression level of miR-889-3p decreased gradually with the prolongation of osteogenic induction. (2) The number, size, and color depth of mineralized nodules in the miR-889-3p inhibitor group were significantly higher than those in the miR-889-3p mimic group and miR-NC group on days 7 and 14 of osteogenic induction (P < 0.05). (3) Overexpression of miR-889-3p significantly suppressed alkaline phosphatase activity, Runx2, osteocalcin, osteopontin, Osterix mRNA and protein expression, whereas the knockdown of miR-889-3p remarkably enhanced osteogenesis-related mRNA and protein expression. (4) The results of dual-luciferase reporter gene assay showed that overexpression of miR-889-3p significantly reduced the luciferase activity of wild-type Runx2. (5) These results suggest that miR-889-3p negatively regulates osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting Runx2.

Key words: miR-889-3p, Runx2, adipose-derived mesenchymal stem cell, osteogenic differentiation, bone defect

CLC Number:

Zhang Yaotian, Cui Jun, Liu Jingyi. miR-889-3p inhibits osteogenic differentiation of adipose-derived mesenchymal stem cells by targeting Runx2[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(19): 3023-3028.

Figures/Tables 6

References

[1] KOONS GL, DIBA M, MIKOS AG. Materials design for bonetissue engineering. Nat Rev Mater. 2020;5(8):584-603.
[2] 周思佳,姜文学,尤佳.骨缺损修复材料：现状与需求和未来[J].中国组织工程研究,2018,22(14):2251-2258.
[3] BHARADWAZ A, JAYASURIYA AC. Recent trends in the application of widely used natural and synthetic polymer nanocomposites in bone tissue regeneration. Mater Sci Eng C Mater Biol Appl. 2020;110: 110698.
[4] ZHANG X, JIANG W, XIE C, et al. Msx1+stem cells recruited by bioactive tissue engineering graft for bone regeneration. Nat Commun. 2022; 13(1):5211.
[5] LI L, WANG Y, WANG Z, et al. Knockdown of FOXA1 enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells partly via activation of the ERK1/2 signalling pathway. Stem Cell Res Ther. 2022;13(1):456.
[6] FELDMAN MD. Editorial Commentary: Free Bone Block With Remplissage Provides Less Translation Than Free Bone Block Alone in Shoulder Instability Patients With Bipolar Bone Loss. Arthroscopy. 2022; 38(9):2618-2619.
[7] CHEN H, SHEN M, SHEN J, et al. A new injectable quick hardening anti-collapse bone cement allows for improving biodegradation and bone repair. Biomater Adv. 2022;141:213098.
[8] YANG C, LIU Y, WANG Z, et al. Controlled mechanical loading improves bone regeneration by regulating type H vessels in a S1Pr1-dependent manner. FASEB J. 2022;36(10):e22530.
[9] XIE C, YE JC, LIANG RJ, et al. Advanced strategies of biomimetic tissue- engineered grafts for bone regeneration.Adv Healthc Mater. 2021; 10(14):2100408.
[10] LEE S, CHAE DS, SONG BW, et al. ADSC-Based Cell Therapies for Musculoskeletal Disorders: A Review of Recent Clinical Trials. Int J Mol Sci. 2021;22(19):10586.
[11] KOMORI T. Regulation of Proliferation, Differentiation and Functions of Osteoblasts by Runx2. Int J Mol Sci. 2019;20(7):1694.
[12] KOMORI T. Molecular Mechanism of Runx2-Dependent Bone Development. Mol Cells. 2020;43(2):168-175.
[13] PARK JS, KIM D, HONG HS. Priming with a Combination of FGF2 and HGF Restores the Impaired Osteogenic Differentiation of Adipose-Derived Stem Cells. Cells. 2022;11(13):2042.
[14] CELIK N, KIM MH, YEO M, et al. miRNA induced 3D bioprinted-heterotypic osteochondral interface. Biofabrication. 2022;14(4). doi: 10.1088/1758-5090/ac7fbb.
[15] SUN X, YANG S, TONG S, et al. Study on Exosomes Promoting the Osteogenic Differentiation of ADSCs in Graphene Porous Titanium Alloy Scaffolds. Front Bioeng Biotechnol. 2022;10:905511.
[16] AGAREVA M, STAFEEV I, MICHURINA S, et al. Type 2 Diabetes Mellitus Facilitates Shift of Adipose-Derived Stem Cells Ex Vivo Differentiation toward Osteogenesis among Patients with Obesity. Life (Basel). 2022; 12(5):688.
[17] ROOHANI I, NO YJ, ZUO B, et al. Low-Temperature Synthesis of Hollow β-Tricalcium Phosphate Particles for Bone Tissue Engineering Applications. ACS Biomater Sci Eng. 2022; 8(5):1806-1815.
[18] ZHANG Y, ZHOU L, ZHANG Z, et al. miR-10a-5p inhibits osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Mol Med Rep. 2020;22(1):135-144.
[19] GE D, CHEN H, ZHENG S, et al. Hsa-miR-889-3p promotes the proliferation of osteosarcoma through inhibiting myeloid cell nuclear differentiation antigen expression. Biomed Pharmacother. 2019;114: 108819.
[20] ZAKRZEWSKI W, DOBRZYNSKI M, SZYMONOWICZ M, et al. Stem cells: Past, present, and future. Stem Cell Res Ther. 2019;10(1):68.
[21] MAQSOOD M, KANG M, WU X, et al. Adult mesenchymal stem cells and their exosomes: Sources, characteristics, and application in regenerative medicine. Life Sci. 2020;256:118002.
[22] FRIEDENSTEIN AJ, CHAILAKHJAN RK, LALYKINA KS. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet. 1970;3:393-403.
[23] ZUK PA, ZHU M, ASHJIAN P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002;13:4279-4295.
[24] MASTROLIA I, FOPPIANI EM, MURGIA A, et al. Challenges in Clinical Development of Mesenchymal Stromal/ Stem Cells: Concise Review. Stem Cells Transl Med. 2019;8(11):1135-1148.
[25] TSUJI W, RUBIN JP, MARRA KG. Adipose-derived stem cells: Implications in tissue regeneration. World J Stem Cells. 2014;6:312-321.
[26] FU X, LIU G, HALIM A, et al. Mesenchymal Stem Cell Migration and Tissue Repair. Cells. 2019;8(8):784.
[27] LU TX, ROTHENBERG ME. MicroRNA. J Allergy Clin Immunol. 2018; 141(4):1202-1207.
[28] TIWARI A, MUKHERJEE B, DIXIT M. MicroRNA Key to Angiogenesis Regulation: MiRNA Biology and Therapy. Curr Cancer Drug Targets. 2018;18(3):266-277.
[29] CHEN L, HEIKKINEN L, WANG C, et al. Trends in the development of miRNA bioinformatics tools. Brief Bioinform. 2019;20(5):1836-1852.
[30] CORREIA DE SOUSA M, GJORGJIEVA M, DOLICKA D, et al. Deciphering miRNAs’ Action through miRNA Editing. Int J Mol Sci. 2019;20(24): 6249.
[31] YANG JX, XIE P, LI YS, et al. Osteoclast-derived miR-23a-5p-containing exosomes inhibit osteogenic differentiation by regulating Runx2. Cell Signal. 2020;70:109504.
[32] REN LR, YAO RB, WANG SY, et al. MiR-27a-3p promotes the osteogenic differentiation by activating CRY2/ERK1/2 axis. Mol Med. 2021;27(1):43.
[33] LI CL, WANG K, LI ZW, et al. MiR-105 enhances osteogenic differentiation of hADSCs via the targeted regulation of SOX9. Tissue Cell. 2021;72:101540.
[34] ZHOU Y, QIAO H, LIU L, et al. miR-21 regulates osteogenic and adipogenic differentiation of BMSCs by targeting PTEN. J Musculoskelet Neuronal Interact. 2021;21(4):568-576.
[35] YIN N, ZHU L, DING L, et al. MiR-135-5p promotes osteoblast differentiation by targeting HIF1AN in MC3T3-E1 cells. Cell Mol Biol Lett. 2019;24:51.
[36] LIU X, XIE S, ZHANG J, et al. Long Noncoding RNA XIST Contributes to Cervical Cancer Development Through Targeting miR-889-3p/SIX1 Axis. Cancer Biother Radiopharm. 2020;35(9):640-649.
[37] JIN Y, XU L, ZHAO B, et al. Tumour-suppressing functions of the lncRNA MBNL1-AS1/miR-889-3p/KLF9 axis in human breast cancer cells. Cell Cycle. 2022;21(9):908-920.
[38] ZHU Q, LI Y, LI L, et al. MicroRNA-889-3p restrains the proliferation and epithelial-mesenchymal transformation of lung cancer cells via down-regulation of Homeodomain-interacting protein kinase 1. Bioengineered. 2021;12(2):10945-10958.
[39] ZHAO X, DONG W, LUO G, et al. Silencing of hsa_circ_0009035 Suppresses Cervical Cancer Progression and Enhances Radiosensitivity through MicroRNA 889-3p-Dependent Regulation of HOXB7. Mol Cell Biol. 2021;41(6):e0063120.
[40] LIU H, XUE Q, CAI H, et al. RUNX3-mediated circDYRK1A inhibits glutamine metabolism in gastric cancer by up-regulating microRNA-889-3p-dependent FBXO4. J Transl Med. 2022;20(1):120.
[41] ZHANG X, WANG P, YUAN K, et al. Hsa_circ_0024093 accelerates VSMC proliferation via miR-4677-3p/miR-889-3p/USP9X/YAP1 axis in vitro model of lower extremity ASO. Mol Ther Nucleic Acids. 2021;26: 511-522.
[42] ZHANG L, ZENG M, TANG F, et al. Circ-PNPT1 contributes to gestational diabetes mellitus (GDM) by regulating the function of trophoblast cells through miR-889-3p/PAK1 axis. Diabetol Metab Syndr. 2021;13(1):58.
[43] NARAYANAN A, SRINAATH N, ROHINI M, et al. Regulation of Runx2 by MicroRNAs in osteoblast differentiation. Life Sci. 2019;232:116676.
[44] CHEN D, KIM DJ, SHEN J, et al. Runx2 plays a central role in Osteoarthritis development. J Orthop Translat. 2019;23:132-139.
[45] GOMATHI K, AKSHAYA N, SRINAATH N, et al. Regulation of Runx2 by post-translational modifications in osteoblast differentiation. Life Sci. 2020;245:117389.