Effect of micro RNA-335-5p regulating bone morphogenetic protein 2 on osteogenic differentiation of human bone marrow mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

HUANG Zhenming ¹ , CAI Zhuo ² , QIAN Jing ¹ , WANG Jianxiong ¹ ,  HU Ning ³

1. Department of Trauma Surgery, the Ninth Hospital of Wuhan City, Wuhan Hubei, 430080, P.R.China;
2. Department of Orthopaedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Hubei, 430000, P.R.China;
3. Department of Orthopaedics, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400000, P.R.China;

Corresponding author：

HU Ning, Email: 1276321387@qq.com

Keywords：

Micro RNA-335-5p; bone morphogenetic protein 2; human bone marrow mesenchymal stem cells; osteogenic differentiation

DOI：

10.7507/1002-1892.201910097

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the effect of micro RNA (miR)-335-5p regulating bone morphogenetic protein 2 (BMP-2) on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs).Methods hBMSCs were cultured in vitro and randomly divided into control group (group A), miR-335-5p mimics group (group B), miR-335-5p mimics negative control group (group C), miR-335-5p inhibitor group (group D), and miR-335-5p inhibitor negative control group (group E). After grouping treatment and induction of osteogenic differentiation, the osteogenic differentiation of cells in each group was detected by alkaline phosphatase (ALP) and alizarin red staining; the expressions of miR-335-5p and BMP-2, Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) mRNAs were detected by real-time fluorescence quantitative PCR analysis; the expressions of Runx2, OPN, OCN, and BMP-2 proteins were detected by Western blot.Results Compared with group A, the relative proportion of ALP positive cells and the relative content of mineralized nodules, the relative expressions of BMP-2, miR-335-5p, OPN, OCN, Runx2 mRNAs, the relative expressions of Runx2, OPN, OCN, and BMP-2 proteins in group B were significantly increased (P<0.05); the above indexes in group D were significantly decreased (P<0.05); the above indexes between groups C, E and group A were not significantly different (P>0.05).Conclusion miR-335-5p can up-regulate BMP-2 expression and promote osteogenic differentiation of hBMSCs.

Citation： HUANG Zhenming, CAI Zhuo, QIAN Jing, WANG Jianxiong, HU Ning. Effect of micro RNA-335-5p regulating bone morphogenetic protein 2 on osteogenic differentiation of human bone marrow mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2020, 34(6): 781-786. doi: 10.7507/1002-1892.201910097 Copy

1.	Zhang J, Tao Z, Wang Y. Long non-coding RNA DANCR regulates the proliferation and osteogenic differentiation of human bone-derived marrow mesenchymal stem cells via the p38 MAPK pathway. Int J Mol Med, 2018, 41(1): 213-219.
2.	代志鹏, 郑稼, 高延征, 等. 谷胱甘肽在激素诱导的 BMSCs 功能失调中的作用. 中国修复重建外科杂志, 2018, 32(1): 91-98.
3.	Jing H, Su X, Gao B, et al. Epigenetic inhibition of Wnt pathway suppresses osteogenic differentiation of BMSCs during osteoporosis. Cell Death Dis, 2018, 9(2): 176.
4.	康明, 黄杰华, 张理选, 等. 壳聚糖/胡须/磷酸钙骨水泥复合生物材料的力学性能及对诱导多能干细胞成骨潜能的影响. 中国修复重建外科杂志, 2018, 32(7): 959-967.
5.	Lin X, Wu L, Zhang Z, et al. MiR-335-5p promotes chondrogenesis in mouse mesenchymal stem cells and is regulated through two positive feedback loops. J Bone Miner Res, 2014, 29(7): 1575-1585.
6.	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.
7.	李济伶, 冯正平, 陈力学, 等. miR-335-5p 对高糖状态下成骨细胞功能的影响. 中华内分泌代谢杂志, 2015, 31(8): 712-716.
8.	An G, Zhang WB, Ma DK, et al. Influence of VEGF/BMP-2 on the proliferation and osteogenetic differentiation of rat bone mesenchymal stem cells on PLGA/gelatin composite scaffold. Eur Rev Med Pharmacol Sci, 2017, 21(10): 2316-2328.
9.	Terauchi M, Inada T, Kanemaru T, et al. Potentiating bioactivity of BMP-2 by polyelectrolyte complexation with sulfonated polyrotaxanes to induce rapid bone regeneration in a mouse calvarial defect. J Biomed Mater Res A, 2017, 105(5): 1355-1363.
10.	Jin C, Zhang P, Zhang M, et al. Inhibition of SLC7A11 by sulfasalazine enhances osteogenic differentiation of mesenchymal stem cells by modulating BMP2/4 expression and suppresses bone loss in ovariectomized mice. J Bone Miner Res, 2017, 32(3): 508-521.
11.	刘红霞, 施琼, 安利钦, 等. miR-155 通过下调 BMP9/Smad 信号通路抑制间充质干细胞 C3H10T1/2 成骨分化. 中国细胞生物学学报, 2017, 39(4): 410-418.
12.	Huang YF, Li LJ, Gao SQ, et al. Evidence based anti-osteoporosis effects of Periplaneta americana L on osteoblasts, osteoclasts, vascular endothelial cells and bone marrow derived mesenchymal stem cells. BMC Complement Altern Med, 2017, 17(1): 413.
13.	Li H, Zhang S, Nie B, et al. The antimicrobial peptide KR-12 promotes the osteogenic differentiation of human bone marrow stem cells by stimulating BMP/SMAD signaling. Rsc Adv, 2018, 8(28): 15547-15557.
14.	Moya A, Larochette N, Bourguignon M, et al. Osteogenic potential of adipogenic predifferentiated human bone marrow-derived multipotent stromal cells for bone tissue-engineering. J Tissue Eng Regen Med, 2018, 12(3): e1511-e1524.
15.	Kocijan R, Muschitz C, Geiger E, et al. Circulating microRNA signatures in patients with idiopathic and postmenopausal osteoporosis and fragility fractures. J Clin Endocrinol Metab, 2016, 101(11): 4125-4134.
16.	何伟珍, 尹志华, 郭佩, 等. 骨关节炎患者关节液中 miR-335-5p 表达及意义. 新医学, 2017, 48(1): 37-39.
17.	汪艳, 吴婷婷, 李静, 等. 釉基质蛋白诱导 MC3T3-E1 细胞成骨分化过程中微小核糖核酸的表达. 口腔材料器械杂志, 2014, 23(4): 198-203.
18.	Sun T, Qu Y, Cui W, et al. Evaluation of osteogenic inductivity of a novel BMP2-mimicking peptide P28 and P28-containing bone composite. J Biomed Mater Res A, 2018, 106(1): 210-220.
19.	Huang H, Dou L, Song J, et al. CBFA2T2 is required for BMP-2-induced osteogenic differentiation of mesenchymal stem cells. Biochem Biophys Res Commun, 2018, 496(4): 1095-1101.
20.	Wang S, Hu S, Wang J, et al. Conditioned medium from bone marrow-derived mesenchymal stem cells inhibits vascular calcification through blockade of the BMP2-Smad1/5/8 signaling pathway. Stem Cell Res Ther, 2018, 9(1): 160.

1. Zhang J, Tao Z, Wang Y. Long non-coding RNA DANCR regulates the proliferation and osteogenic differentiation of human bone-derived marrow mesenchymal stem cells via the p38 MAPK pathway. Int J Mol Med, 2018, 41(1): 213-219.
2. 代志鹏, 郑稼, 高延征, 等. 谷胱甘肽在激素诱导的 BMSCs 功能失调中的作用. 中国修复重建外科杂志, 2018, 32(1): 91-98.
3. Jing H, Su X, Gao B, et al. Epigenetic inhibition of Wnt pathway suppresses osteogenic differentiation of BMSCs during osteoporosis. Cell Death Dis, 2018, 9(2): 176.
4. 康明, 黄杰华, 张理选, 等. 壳聚糖/胡须/磷酸钙骨水泥复合生物材料的力学性能及对诱导多能干细胞成骨潜能的影响. 中国修复重建外科杂志, 2018, 32(7): 959-967.
5. Lin X, Wu L, Zhang Z, et al. MiR-335-5p promotes chondrogenesis in mouse mesenchymal stem cells and is regulated through two positive feedback loops. J Bone Miner Res, 2014, 29(7): 1575-1585.
6. 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.
7. 李济伶, 冯正平, 陈力学, 等. miR-335-5p 对高糖状态下成骨细胞功能的影响. 中华内分泌代谢杂志, 2015, 31(8): 712-716.
8. An G, Zhang WB, Ma DK, et al. Influence of VEGF/BMP-2 on the proliferation and osteogenetic differentiation of rat bone mesenchymal stem cells on PLGA/gelatin composite scaffold. Eur Rev Med Pharmacol Sci, 2017, 21(10): 2316-2328.
9. Terauchi M, Inada T, Kanemaru T, et al. Potentiating bioactivity of BMP-2 by polyelectrolyte complexation with sulfonated polyrotaxanes to induce rapid bone regeneration in a mouse calvarial defect. J Biomed Mater Res A, 2017, 105(5): 1355-1363.
10. Jin C, Zhang P, Zhang M, et al. Inhibition of SLC7A11 by sulfasalazine enhances osteogenic differentiation of mesenchymal stem cells by modulating BMP2/4 expression and suppresses bone loss in ovariectomized mice. J Bone Miner Res, 2017, 32(3): 508-521.
11. 刘红霞, 施琼, 安利钦, 等. miR-155 通过下调 BMP9/Smad 信号通路抑制间充质干细胞 C3H10T1/2 成骨分化. 中国细胞生物学学报, 2017, 39(4): 410-418.
12. Huang YF, Li LJ, Gao SQ, et al. Evidence based anti-osteoporosis effects of Periplaneta americana L on osteoblasts, osteoclasts, vascular endothelial cells and bone marrow derived mesenchymal stem cells. BMC Complement Altern Med, 2017, 17(1): 413.
13. Li H, Zhang S, Nie B, et al. The antimicrobial peptide KR-12 promotes the osteogenic differentiation of human bone marrow stem cells by stimulating BMP/SMAD signaling. Rsc Adv, 2018, 8(28): 15547-15557.
14. Moya A, Larochette N, Bourguignon M, et al. Osteogenic potential of adipogenic predifferentiated human bone marrow-derived multipotent stromal cells for bone tissue-engineering. J Tissue Eng Regen Med, 2018, 12(3): e1511-e1524.
15. Kocijan R, Muschitz C, Geiger E, et al. Circulating microRNA signatures in patients with idiopathic and postmenopausal osteoporosis and fragility fractures. J Clin Endocrinol Metab, 2016, 101(11): 4125-4134.
16. 何伟珍, 尹志华, 郭佩, 等. 骨关节炎患者关节液中 miR-335-5p 表达及意义. 新医学, 2017, 48(1): 37-39.
17. 汪艳, 吴婷婷, 李静, 等. 釉基质蛋白诱导 MC3T3-E1 细胞成骨分化过程中微小核糖核酸的表达. 口腔材料器械杂志, 2014, 23(4): 198-203.
18. Sun T, Qu Y, Cui W, et al. Evaluation of osteogenic inductivity of a novel BMP2-mimicking peptide P28 and P28-containing bone composite. J Biomed Mater Res A, 2018, 106(1): 210-220.
19. Huang H, Dou L, Song J, et al. CBFA2T2 is required for BMP-2-induced osteogenic differentiation of mesenchymal stem cells. Biochem Biophys Res Commun, 2018, 496(4): 1095-1101.
20. Wang S, Hu S, Wang J, et al. Conditioned medium from bone marrow-derived mesenchymal stem cells inhibits vascular calcification through blockade of the BMP2-Smad1/5/8 signaling pathway. Stem Cell Res Ther, 2018, 9(1): 160.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of micro RNA-335-5p regulating bone morphogenetic protein 2 on osteogenic differentiation of human bone marrow mesenchymal stem cells

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