MicroRNA-98-5p promotes osteoblast proliferation and differentiation: possibilities and mechanisms

doi:10.12307/2021.107

Abstract

Abstract:

BACKGROUND: MicroRNA-98-5p (miR-98-5p) can inhibit the high mobility group AT-HOOK 2 (HMGA2). The phosphatidylinositol 3-kinase/alkaline phosphatase/glycogen synthase kinase-3β (PI3K/AKT/GSK-3β) signaling pathway is involved in cell proliferation. During the fracture healing process, it is unclear whether miR-98-5p can act on the PI3K/Akt/GSK-3β pathway by activating HMGA2 to promote the proliferation of osteoblasts.

OBJECTIVE: To investigate the mechanisms of microRNA-98-5p (miR-98-5p) in promoting osteoblast proliferation and differentiation by regulating HMGA2 and PI3K/Akt/GSK-3β pathway.
METHODS: MC3T3-E1 cells transfected with miR-98-5p mimics and HMGA2 plasmid by Lipofectamine 2000 were divided into a miR-98-5p mimics group and a HMGA2 overexpression group, respectively. MC3T3-E1 cells transfected with dimethyl sulfoxide and scrambled plasmids were divided into a mimic control group and a scramble group. Cells transiently transfected with miR-98-5p inhibitor using liposomes were as a miR-98-5p inhibitor group. Normal MC3T3-E1 cells without treatment were used as a blank control group.
RESULTS AND CONCLUSION: There were no significant differences in HMGA2, PI3K/Akt/GSK-3β, alkaline phosphatase activity and mRNA level between the blank control group and the mimic control group. Compared with the mimic control group, the HMGA2 and PI3K/Akt/GSK-3β protein expression, cell differentiation and proliferation, alkaline phosphatase activity and mRNA level were significantly reduced in the miR-98-5p mimics group. The interaction between miR-98-5p and HMGA2 predicted by Targetscan7.1 showed that compared with the mimic control group, the HMGA2 protein and mRNA were reduced in the miR-98-5p mimic group, while compared with the miR-98-5p mimic group, the HMGA2 protein and mRNA had an increase in the miR-98-5p inhibitor group. There were no significant differences in osteoblast proliferation, alkaline phosphatase activity and mRNA between the blank control group and the scramble group. Compared with the scramble group, the PI3K/Akt/GSK-3β expression, cell differentiation and proliferation, alkaline phosphatase activity and mRNA in the HMGA2 overexpression group were significantly increased. To conclude, miR-98-5p can inhibit the HMGA2 and PI3K/Akt/GSK-3β expressions in MC3T3-E1 cells, inhibit cell proliferation and differentiation. HMGA2 is possible the direct target of miR-98-5p.

Key words: osteoblasts, microRNA-98-5p, high mobility group AT-HOOK 2, phosphatidylinositol 3-kinase, protein kinase B, glycogen synthase kinase 3β, transfection, MC3T3-E1 cells, proliferation, differentiation

CLC Number:

Zheng Feng, Zhang Fucai, Xu Zhe. MicroRNA-98-5p promotes osteoblast proliferation and differentiation: possibilities and mechanisms[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(26): 4112-4117.

Figures/Tables 7

References

[1] Qiao Z, Greven J, Horst K, et al. Fracture Healing and the Underexposed Role of Extracellular Vesicle-Based Cross Talk. Shock. 2018;49(5):486-496.
[2] Bergdolt S, Kovtun A, Hägele Y, et al. Osteoblast-specific overexpression of complement receptor C5aR1 impairs fracture healing. PLoS One. 2017;12(6):e0179512.
[3] Sekiguchi Y, Mano H, Nakatani S, et al. Mangiferin positively regulates osteoblast differentiation and suppresses osteoclast differentiation. Mol Med Rep. 2017;16(2):1328-1332.
[4] Chen X, Wang Z, Duan N, et al. Osteoblast-osteoclast interactions. Connect Tissue Res. 2018;59(2):99-107.
[5] Jiao H, Xiao E, Graves DT. Diabetes and Its Effect on Bone and Fracture Healing. Curr Osteoporos Rep. 2015;13(5):327-335.
[6] Prideaux M, Findlay DM, Atkins GJ. Osteocytes: The master cells in bone remodelling. Curr Opin Pharmacol. 2016;28:24-30.
[7] LEE WC, GUNTUR AR, LONG F, et al. Energy Metabolism of the Osteoblast: Implications for Osteoporosis. Endocr Rev. 2017;38(3):255-266.
[8] Ghiasi MS, Chen J, Vaziri A, et al. Bone fracture healing in mechanobiological modeling: A review of principles and methods. Bone Rep. 2017;6:87-100.
[9] Piccoli MT, Gupta SK, Thum T. Noncoding RNAs as regulators of cardiomyocyte proliferation and death. J Mol Cell Cardiol. 2015;89(Pt A):59-67.
[10] Han T, Wu N, Wang Y, et al. miR‑16‑2‑3p inhibits cell proliferation and migration and induces apoptosis by targeting PDPK1 in maxillary primordium mesenchymal cells. Int J Mol Med. 2019;43(3):1441-1451.
[11] Zhang M, Wang F, Zhang X. miRNA-627 inhibits cell proliferation and cell migration, promotes cell apoptosis in prostate cancer cells through upregulating MAP3K1, PTPRK and SRA1. Int J Clin Exp Pathol. 2018;11(1):255-261.
[12] Qiu K, Xie Q, Jiang S, et al. miR-98-5p promotes apoptosis and inhibits migration and cell growth in papillary thyroid carcinoma through Bax/Caspase-3 by HMGA2. J Clin Lab Anal. 2020;34(2):e23044.
[13] Ma P, Gu B, Xiong W, et al. Glimepiride promotes osteogenic differentiation in rat osteoblasts via the PI3K/Akt/eNOS pathway in a high glucose microenvironment. PLoS One. 2014;9(11):e112243.
[14] Majidinia M, Sadeghpour A, Yousefi B. The roles of signaling pathways in bone repair and regeneration. J Cell Physiol. 2018;233(4): 2937-2948.
[15] Zhu Y, Xu J, Liang W, et al. miR-98-5p Alleviated Epithelial-to-Mesenchymal Transition and Renal Fibrosis via Targeting Hmga2 in Diabetic Nephropathy. Int J Endocrinol. 2019;2019:4946181.
[16] Yuan H, Zhao H, Wang J, et al. MicroRNA let-7c-5p promotes osteogenic differentiation of dental pulp stem cells by inhibiting lipopolysaccharide-induced inflammation via HMGA2/PI3K/Akt signal blockade. Clin Exp Pharmacol Physiol. 2019;46(4):389-397.
[17] Hu C, Zhang T, Ren B, et al. Effect of vacuum-assisted closure combined with open bone grafting to promote rabbit bone graft vascularization. Med Sci Monit. 2015;21:1200-1206.
[18] Hammond SM. An overview of microRNAs. Adv Drug Deliv Rev. 2015; 87:3-14.
[19] Soifer HS, Rossi JJ, Saetrom P. MicroRNAs in disease and potential therapeutic applications. Mol Ther. 2007;15(12):2070-2079.
[20] Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet. 2010;11(9): 597-610.
[21] Tang J, Zhang Z, Jin X, et al. miR-383 negatively regulates osteoblastic differentiation of bone marrow mesenchymal stem cells in rats by targeting Satb2. Bone. 2018;114:137-143.
[22] Arfat Y, Basra MAR, Shahzad M, et al. miR-208a-3p Suppresses Osteoblast Differentiation and Inhibits Bone Formation by Targeting ACVR1. Mol Ther Nucleic Acids. 2018;11:323-336.
[23] Zhang L, Tang Y, Zhu X, et al. Overexpression of MiR-335-5p Promotes Bone Formation and Regeneration in Mice. J Bone Miner Res. 2017; 32(12):2466-2475.
[24] Kureel J, John AA, Dixit M, et al. MicroRNA-467g inhibits new bone regeneration by targeting Ihh/Runx-2 signaling. Int J Biochem Cell Biol. 2017;85:35-43.
[25] Zhang Y, Gao Y, Cai L, et al. MicroRNA-221 is involved in the regulation of osteoporosis through regulates RUNX2 protein expression and osteoblast differentiation. Am J Transl Res. 2017;9(1):126-135.
[26] Chen G, Deng C, Li YP. TGF-β and BMP signaling in osteoblast differentiation and bone formation. Int J Biol Sci. 2012;8(2):272-288.
[27] Bertacchini J, Heidari N, Mediani L, et al. Targeting PI3K/AKT/mTOR network for treatment of leukemia. Cell Mol Life Sci. 2015; 72(12):2337-2347.
[28] Manfredi GI, Dicitore A, Gaudenzi G, et al. PI3K/Akt/mTOR signaling in medullary thyroid cancer: a promising molecular target for cancer therapy. Endocrine. 2015;48(2):363-370.
[29] Wang J, Ma XY, Feng YF, et al. Magnesium Ions Promote the Biological Behaviour of Rat Calvarial Osteoblasts by Activating the PI3K/Akt Signalling Pathway. Biol Trace Elem Res. 2017;179(2):284-293.
[30] Ma P, Gu B, Ma J, et al. Glimepiride induces proliferation and differentiation of rat osteoblasts via the PI3-kinase/Akt pathway. Metabolism. 2010;59(3):359-366.
[31] Phukan S, Babu VS, Kannoji A, et al. GSK3beta: role in therapeutic landscape and development of modulators. Br J Pharmacol. 2010; 160(1):1-19.
[32] Guo B, Zhang W, Xu S, et al. GSK-3β mediates dexamethasone-induced pancreatic β cell apoptosis. Life Sci. 2016;144:1-7.
[33] Sharma M, Chuang WW, Sun Z. Phosphatidylinositol 3-kinase/Akt stimulates androgen pathway through GSK3beta inhibition and nuclear beta-catenin accumulation. J Biol Chem. 2002;277(34):30935-30941.
[34] Yao CJ, Lv Y, Zhang CJ, et al. MicroRNA-185 inhibits the growth and proliferation of osteoblasts in fracture healing by targeting PTH gene through down-regulating Wnt/β -catenin axis: In an animal experiment. Biochem Biophys Res Commun. 2018;501(1):55-63.
[35] Song F, Jiang D, Wang T, et al. Mechanical Stress Regulates Osteogenesis and Adipogenesis of Rat Mesenchymal Stem Cells through PI3K/Akt/GSK-3β/β-Catenin Signaling Pathway. Biomed Res Int. 2017;2017:6027402.
[36] Zheng X, Dai J, Zhang H, et al. MicroRNA-221 promotes cell proliferation, migration, and differentiation by regulation of ZFPM2 in osteoblasts. Braz J Med Biol Res. 2018;51(12):e7574.
[37] Zheng F, Wang F, Xu Z. MicroRNA-98-5p prevents bone regeneration by targeting high mobility group AT-Hook 2. Exp Ther Med. 2019;18(4): 2660-2666.
[38] 刘珍星,张山锋,杨钟华.IL-38通过调控PI3K/Akt/GSK3β/NFATc1信号通路抑制骨质疏松的机制研究[J].中国免疫学杂志,2018,34(2): 251-255.
[39] Guo Z, He C, Yang F, et al. Long non-coding RNA-NEAT1, a sponge for miR-98-5p, promotes expression of oncogene HMGA2 in prostate cancer. Biosci Rep. 2019;39(9):BSR20190635.