Overexpression of miRNA-22-3p attenuates osteoporosis by targeting MAPK14

Jia,Xiaolin; Yang,Ming; Hu,Wei; Cai,San

doi:10.3892/etm.2021.10124

Overexpression of miRNA-22-3p attenuates osteoporosis by targeting MAPK14

Authors:
- Xiaolin Jia
- Ming Yang
- Wei Hu
- San Cai
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Affiliations: Department of Orthopaedics, Chongqing General Hospital, Chongqing 401147, P.R. China, Department of Orthopaedics, Chongqing Public Health Medical Center, Chongqing 400036, P.R. China
Published online on: May 2, 2021 https://doi.org/10.3892/etm.2021.10124
Article Number: 692
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Osteoporosis (OP) results from an imbalance between bone formation, which is regulated by osteoblasts, and bone resorption, which is mediated by osteoclasts. MicroRNA-22-3p (miR-22-3p) expression is decreased during the process of osteoclast differentiation and p38α mitogen-activated protein kinase (MAPK)14 promotes the proliferation and differentiation of osteoclast progenitors. However, whether miR-22-3p could target MAPK14 to regulate the progression of OP remains unknown, which was the aim of the present study. CD14⁺ PBMCs were used for the establishment of osteoclastic differentiation in vitro. In the present study, reverse transcription quantitative PCR was used to determine the mRNA expression of MAPK14, tartrate resistant acid phosphatase (TRAP), nuclear factor of activated T-cells (NFATC1) and cathepsin K (CTSK). Western blotting was applied to determine the protein expression of MAPK14, TRAP, NFATC1, CTSK, p-p65 and p65. Dual luciferase reporter assay was applied to confirm the relation between miR-22-3p and MAPK14. Cell Counting Kit-8 assay and flow cytometry assays were used to determine the cell proliferation and cell apoptosis, respectively. The results demonstrated that miR-22-3p expression was lower while MAPK14 expression was higher in the serum from patients with OP compared with healthy volunteers. Furthermore, miR-22-3p expression was negatively correlated with MAPK14 expression in patients with OP. In addition, miR-22-3p expression was decreased and MAPK14 expression was increased during the progression of CD14⁺peripheral blood mononuclear cells (PBMCs) osteoclastic differentiation in a time-dependent manner. Furthermore, miR-22-3p inhibited the proliferation and differentiation and promoted the apoptosis of CD14⁺PBMCs by targeting MAPK14. In summary, the findings from the present study suggested that miR-22-3p may serve a potential therapeutic role in patients with OP.

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1	Cosman F, de Beur SJ, LeBoff MS, Lewiecki EM, Tanner B, Randall S and Lindsay R: Erratum to: Clinician's guide to prevention and treatment of osteoporosis. Osteoporos Int. 26:2045–2047. 2015.PubMed/NCBI View Article : Google Scholar
2	Nguyen BN, Hoshino H, Togawa D and Matsuyama Y: Cortical thickness index of the proximal femur: A radiographic parameter for preliminary assessment of bone mineral density and osteoporosis status in the age 50 years and over population. Clin Orthop Surg. 10:149–156. 2018.PubMed/NCBI View Article : Google Scholar
3	Xiao W, Wang Y, Pacios S, Li S and Graves DT: Cellular and molecular aspects of bone remodeling. Front Oral Biol. 18:9–16. 2016.PubMed/NCBI View Article : Google Scholar
4	Karsenty G: Transcriptional control of skeletogenesis. Annu Rev Genomics Hum Genet. 9:183–196. 2008.PubMed/NCBI View Article : Google Scholar
5	Teitelbaum SL: Bone resorption by osteoclasts. Science. 289:1504–1508. 2000.PubMed/NCBI View Article : Google Scholar
6	Manolagas SC: From estrogen-centric to aging and oxidative stress: A revised perspective of the pathogenesis of osteoporosis. Endocr Rev. 31:266–300. 2010.PubMed/NCBI View Article : Google Scholar
7	Rachner TD, Khosla S and Hofbauer LC: Osteoporosis: Now and the future. Lancet. 377:1276–1287. 2011.PubMed/NCBI View Article : Google Scholar
8	Landgraf P, Rusu M, Sheridan R, Sewer A, Iovino N, Aravin A, Pfeffer S, Rice A, Kamphorst AO, Landthaler M, et al: A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 129:1401–1414. 2007.PubMed/NCBI View Article : Google Scholar
9	Gámez B, Rodriguez-Carballo E and Ventura F: MicroRNAs and post-transcriptional regulation of skeletal development. J Mol Endocrinol. 52:R179–R197. 2014.PubMed/NCBI View Article : Google Scholar
10	Lian JB, Stein GS, van Wijnen AJ, Stein JL, Hassan MQ, Gaur T and Zhang Y: MicroRNA control of bone formation and homeostasis. Nat Rev Endocrinol. 8:212–227. 2012.PubMed/NCBI View Article : Google Scholar
11	Mizuno Y, Yagi K, Tokuzawa Y, Kanesaki-Yatsuka Y, Suda T, Katagiri T, Fukuda T, Maruyama M, Okuda A, Amemiya T, et al: miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun. 368:267–272. 2008.PubMed/NCBI View Article : Google Scholar
12	Wang FS, Chuang PC, Lin CL, Chen MW, Ke HJ, Chang YH, Chen YS, Wu SL and Ko JY: MicroRNA-29a protects against glucocorticoid-induced bone loss and fragility in rats by orchestrating bone acquisition and resorption. Arthritis Rheum. 65:1530–1540. 2013.PubMed/NCBI View Article : Google Scholar
13	Kahai S, Lee SC, Lee DY, Yang J, Li M, Wang CH, Jiang Z, Zhang Y, Peng C and Yang BB: MicroRNA miR-378 regulates nephronectin expression modulating osteoblast differentiation by targeting GalNT-7. PLoS One. 4(e7535)2009.PubMed/NCBI View Article : Google Scholar
14	Xia Z, Chen C, Chen P, Xie H and Luo X: MicroRNAs and their roles in osteoclast differentiation. Front Med. 5:414–419. 2011.PubMed/NCBI View Article : Google Scholar
15	van Wijnen AJ, van de Peppel J, van Leeuwen JP, Lian JB, Stein GS, Westendorf JJ, Oursler MJ, Im HJ, Taipaleenmäki H, Hesse E, et al: MicroRNA functions in osteogenesis and dysfunctions in osteoporosis. Curr Osteoporos Rep. 11:72–82. 2013.PubMed/NCBI View Article : Google Scholar
16	Weilner S, Skalicky S, Salzer B, Keider V, Wagner M, Hildner F, Gabriel C, Dovjak P, Pietschmann P, Grillari-Voglauer R, et al: Differentially circulating miRNAs after recent osteoporotic fractures can influence osteogenic differentiation. Bone. 79:43–51. 2015.PubMed/NCBI View Article : Google Scholar
17	Ma Y, Shan Z, Ma J, Wang Q, Chu J, Xu P, Qin A and Fan S: Validation of downregulated microRNAs during osteoclast formation and osteoporosis progression. Mol Med Rep. 13:2273–2280. 2016.PubMed/NCBI View Article : Google Scholar
18	Mäkitie RE, Hackl M, Niinimäki R, Kakko S, Grillari J and Mäkitie O: Altered microRNA profile in osteoporosis caused by impaired WNT signaling. J Clin Endocrinol Metab. 103:1985–1996. 2018.PubMed/NCBI View Article : Google Scholar
19	Zhang X, Wang Y, Zhao H, Han X, Zhao T, Qu P, Li G and Wang W: Extracellular vesicle-encapsulated miR-22-3p from bone marrow mesenchymal stem cell promotes osteogenic differentiation via FTO inhibition. Stem Cell Res Ther. 11(227)2020.PubMed/NCBI View Article : Google Scholar
20	World Health Organization. Assessment of fracture risk and its application to screening for postmenopausal osteoporosis. Report of a WHO Study Group. World Health Organ Tech Rep Ser. 843:1–129. 1994.PubMed/NCBI View Article : Google Scholar
21	Hemingway F, Cheng X, Knowles HJ, Estrada FM, Gordon S and Athanasou NA: In vitro generation of mature human osteoclasts. Calcif Tissue Int. 89:389–395. 2011.PubMed/NCBI View Article : Google Scholar
22	Sørensen MG, Henriksen K, Schaller S, Henriksen DB, Nielsen FC, Dziegiel MH and Karsdal MA: Characterization of osteoclasts derived from CD14⁺ monocytes isolated from peripheral blood. J Bone Miner Metab. 25:36–45. 2007.PubMed/NCBI View Article : Google Scholar
23	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.PubMed/NCBI View Article : Google Scholar
24	Cheng P, Chen C, He HB, Hu R, Zhou HD, Xie H, Zhu W, Dai RC, Wu XP, Liao EY, et al: miR-148a regulates osteoclastogenesis by targeting V-maf musculoaponeurotic fibrosarcoma oncogene homolog B. J Bone Miner Res. 28:1180–1190. 2013.PubMed/NCBI View Article : Google Scholar
25	Jing D, Hao J, Shen Y, Tang G, Li ML, Huang SH and Zhao ZH: The role of microRNAs in bone remodeling. Int J Oral Sci. 7:131–143. 2015.PubMed/NCBI View Article : Google Scholar
26	García Palacios V, Robinson LJ, Borysenko CW, Lehmann T, Kalla SE and Blair HC: Negative regulation of RANKL-induced osteoclastic differentiation in RAW264.7 cells by estrogen and phytoestrogens. J Biol Chem. 280:13720–13727. 2005.PubMed/NCBI View Article : Google Scholar
27	Sugatani T and Hruska KA: Down-regulation of miR-21 biogenesis by estrogen action contributes to osteoclastic apoptosis. J Cell Biochem. 114:1217–1222. 2013.PubMed/NCBI View Article : Google Scholar
28	Kagiya T and Nakamura S: Expression profiling of microRNAs in RAW264.7 cells treated with a combination of tumor necrosis factor alpha and RANKL during osteoclast differentiation. J Periodontal Res. 48:373–385. 2013.PubMed/NCBI View Article : Google Scholar
29	Shibuya H, Nakasa T, Adachi N, Nagata Y, Ishikawa M, Deie M, Suzuki O and Ochi M: Overexpression of microRNA-223 in rheumatoid arthritis synovium controls osteoclast differentiation. Mod Rheumatol. 23:674–685. 2013.PubMed/NCBI View Article : Google Scholar
30	Cao L, Liu W, Zhong Y, Zhang Y, Gao D, He T, Liu Y, Zou Z, Mo Y, Peng S, et al: Linc02349 promotes osteogenesis of human umbilical cord-derived stem cells by acting as a competing endogenous RNA for miR-25-3p and miR-33b-5p. Cell Prolif. 53(e12814)2020.PubMed/NCBI View Article : Google Scholar
31	Cargnello M and Roux PP: Activation and function of the MAPKs and their substrates, the MAPK-activated protein kinases. Microbiol Mol Biol Rev. 75:50–83. 2011.PubMed/NCBI View Article : Google Scholar
32	Cuadrado A and Nebreda AR: Mechanisms and functions of p38 MAPK signalling. Biochem J. 429:403–417. 2010.PubMed/NCBI View Article : Google Scholar
33	Thamodaran V and Bruce AW: p38 (Mapk14/11) occupies a regulatory node governing entry into primitive endoderm differentiation during preimplantation mouse embryo development. Open Biol. 6(160190)2016.PubMed/NCBI View Article : Google Scholar
34	Caverzasio J, Higgins L and Ammann P: Prevention of trabecular bone loss induced by estrogen deficiency by a selective p38alpha inhibitor. J Bone Miner Res. 23:1389–1397. 2008.PubMed/NCBI View Article : Google Scholar
35	Thouverey C and Caverzasio J: Ablation of p38α MAPK signaling in osteoblast lineage cells protects mice from bone loss induced by estrogen deficiency. Endocrinology. 156:4377–4387. 2015.PubMed/NCBI View Article : Google Scholar
36	Cong Q, Jia H, Li P, Qiu S, Yeh J, Wang Y, Zhang ZL, Ao J, Li B and Liu H: p38α MAPK regulates proliferation and differentiation of osteoclast progenitors and bone remodeling in an aging-dependent manner. Sci Rep. 7(45964)2017.PubMed/NCBI View Article : Google Scholar
37	Boyle WJ, Simonet WS and Lacey DL: Osteoclast differentiation and activation. Nature. 423:337–342. 2003.PubMed/NCBI View Article : Google Scholar
38	Jovicic A, Zaldivar Jolissaint JF, Moser R, Silva Santos MF and Luthi-Carter R: MicroRNA-22 (miR-22) overexpression is neuroprotective via general anti-apoptotic effects and may also target specific Huntington's disease-related mechanisms. PLoS One. 8(e54222)2013.PubMed/NCBI View Article : Google Scholar
39	Kwon M, Kim JM, Lee K, Park SY, Lim HS, Kim T and Jeong D: Synchronized cell cycle arrest promotes osteoclast differentiation. Int J Mol Sci. 17(1292)2016.PubMed/NCBI View Article : Google Scholar
40	Boyce BF: Advances in osteoclast biology reveal potential new drug targets and new roles for osteoclasts. J Bone Miner Res. 28:711–722. 2013.PubMed/NCBI View Article : Google Scholar