Study on adsorption of microRNA-124 by long chain non-coding RNA MALAT1 regulates osteogenic differentiation of mesenchymal stem cells_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Yang ¹ , GUO Hai ² , MA Li ³ ,  ZHU Jinyu ¹ , GUO Anyun ¹ , HE Yong ¹

1. Department of Joint Trauma, General Hospital of Shenzhen University, Shenzhen Guangdong, 518055, P.R.China;
2. Department of Anesthesiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi Xinjiang, 830054, P.R.China;
3. Department of Emergency, General Hospital of Xinjiang Military Region, Urumqi Xinjiang, 830000, P.R.China;

Corresponding author：

ZHU Jinyu, Email: icu55555@163.com

Keywords：

Long chain non-coding RNA; metastasis-associated lung adenocarcinoma transcript 1; microRNA-124; mesenchymal stem cells; osteogenic differentiation

DOI：

10.7507/1002-1892.201906025

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the regulatory effect of long chain non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) adsorbing microRNA-124 (miR-124) on osteogenic differentiation of mesenchymal stem cells (MSCs).Methods C3H10T1/2 cells derived from mouse embryos were cultured in vitro, then randomly divided into control group (group A), lncRNA MALAT1 no-load plasmid group (group B), lncRNA MALAT1 overexpression plasmid group (group C), lncRNA MALAT1 small interfering RNA (siRNA) group (group D), and lncRNA MALAT1 siRNA negative control group (group E). The cells were transfected into plasmids and siRNA, then induced to differentiate into osteoblasts. Alkaline phosphatase (ALP) and alizarin red staining were used to detect the osteogenic differentiation of cells in each group, real-time fluorescence quantitative (qRT-PCR) analysis was used to detect the expressions of lncRNA MALAT, miR-124, and osteogenesis-related genes such as Runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN) in each group. Double luciferase reporter gene was used to detect the targeting regulation of lncRNA MALAT1 to miR-124.Results The relative contents of ALP positive cells, mineralized nodule, and the relative mRNA expressions of lncRNA MALAT1, Runx2, OPN, and OCN in group C were significantly higher than those in other groups (P<0.05), while in group D significantly lower than in other groups (P<0.05); the relative expression of miR-124 in group C was significantly lower than that in other groups(P<0.05), while in group D significantly higher than in other groups (P<0.05). There was no significant difference in these indexes between groups A, B, and E (P>0.05). The results of double luciferase reporter gene assay showed that lncRNA MALAT1 targeting down-regulated the expression of miR-124.Conclusion LncRNA MALAT1 can targeting down-regulate the expression of miR-124 and promote the osteogenic differentiation of MSCs.

Citation： ZHANG Yang, GUO Hai, MA Li, ZHU Jinyu, GUO Anyun, HE Yong. Study on adsorption of microRNA-124 by long chain non-coding RNA MALAT1 regulates osteogenic differentiation of mesenchymal stem cells. Chinese Journal of Reparative and Reconstructive Surgery, 2020, 34(2): 240-245. doi: 10.7507/1002-1892.201906025 Copy

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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.	Lekamwasam S, Chandran M, Subasinghe S. Revised FRAX^®-based intervention thresholds for the management of osteoporosis among postmenopausal women in Sri Lanka. Arch Osteoporos, 2019, 14(1): 33.
13.	Liang X, Du Y, Wen Y, et al. Assessing the genetic correlations between blood plasma proteins and osteoporosis: a polygenic risk score analysis. Calcif Tissue Int, 2019, 104(2): 171-181.
14.	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.
15.	Yi J, Liu D, Xiao J. LncRNA MALAT1 sponges miR-30 to promote osteoblast differentiation of adipose-derived mesenchymal stem cells by promotion of Runx2 expression. Cell Tissue Res, 2019, 376(1): 113-121.
16.	Gao Y, Xiao F, Wang C, et al. Long noncoding RNA MALAT1 promotes osterix expression to regulate osteogenic differentiation by targeting miRNA-143 in human bone marrow-derived mesenchymal stem cells. J Cell Biochem, 2018, 119(8): 6986-6996.
17.	马智勇, 郭成龙. lncRNA 对骨质疏松骨代谢相关信号通路影响的研究进展. 中国骨质疏松杂志, 2018, 24(4): 547-551.
18.	Zhang TH, Liang LZ, Liu XL, et al. Long non-coding RNA MALAT1 interacts with miR-124 and modulates tongue cancer growth by targeting JAG1. Oncol Rep, 2017, 37(4): 2087-2094.
19.	田允鸿, 曾星, 邱慧芝, 等. MiR-124 对鼻咽癌细胞株上皮间充质转化和放疗敏感性的影响. 实用医学杂志, 2016, 32(17): 2780-2783.
20.	郭晓宇, 李唯, 陈克明, 等. 淫羊藿苷通过 PI3K/AKT-eNOS 信号途径促进大鼠骨髓基质细胞的成骨性分化. 中国药理学通报, 2013, 29(7): 966-970.

1. Entezari V, Lazarus M. Surgical considerations in managing osteoporosis, osteopenia, and vitamin D deficiency during arthroscopic rotator cuff repair. Orthop Clin North Am, 2019, 50(2): 233-243.
2. 肖飞, 焦竞, 黄玉成, 等. CD44 分子岩藻糖基化修饰对兔 BMSCs 流体黏附力的影响. 中国修复重建外科杂志, 2018, 32(1): 99-103.
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. Wang Q, Yang Q, Chen G, et al. LncRNA expression profiling of BMSCs in osteonecrosis of the femoral head associated with increased adipogenic and decreased osteogenic differentiation. Sci Rep, 2018, 8(1): 9127-9140.
5. Dong Y, Liang G, Yuan B, et al. MALAT1 promotes the proliferation and metastasis of osteosarcoma cells by activating the PI3K/Akt pathway. Tumour Biol, 2015, 36(3): 1477-1486.
6. Che W, Dong Y, Quan HB. RANKL inhibits cell proliferation by regulating MALAT1 expression in a human osteoblastic cell line hFOB 1.19. Cell Mol Biol (Noisy-le-grand), 2015, 61(1): 7-14.
7. 江春燕, 宋成文. PI3K/AKT 信号通路对骨髓间充质干细胞行为学调节的研究进展. 湖南中医杂志, 2017, 33(12): 176-178.
8. Feng T, Shao F, Wu Q, et al. miR-124 downregulation leads to breast cancer progression via LncRNA-MALAT1 regulation and CDK4/E2F1 signal activation. Oncotarget, 2016, 7(13): 16205-16216.
9. 李慧, 李子英. miR-9 和 miR-124 促进骨髓间充质干细胞向神经细胞分化. 解剖科学进展, 2016, 22(3): 292-294.
10. Zheng YB, Xiao GC, Tong SL, et al. Paeoniflorin inhibits human gastric carcinoma cell proliferation through up-regulation of microRNA-124 and suppression of PI3K/Akt and STAT3 signaling. World J Gastroenterol, 2015, 21(23): 7197-7207.
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. Lekamwasam S, Chandran M, Subasinghe S. Revised FRAX^®-based intervention thresholds for the management of osteoporosis among postmenopausal women in Sri Lanka. Arch Osteoporos, 2019, 14(1): 33.
13. Liang X, Du Y, Wen Y, et al. Assessing the genetic correlations between blood plasma proteins and osteoporosis: a polygenic risk score analysis. Calcif Tissue Int, 2019, 104(2): 171-181.
14. 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.
15. Yi J, Liu D, Xiao J. LncRNA MALAT1 sponges miR-30 to promote osteoblast differentiation of adipose-derived mesenchymal stem cells by promotion of Runx2 expression. Cell Tissue Res, 2019, 376(1): 113-121.
16. Gao Y, Xiao F, Wang C, et al. Long noncoding RNA MALAT1 promotes osterix expression to regulate osteogenic differentiation by targeting miRNA-143 in human bone marrow-derived mesenchymal stem cells. J Cell Biochem, 2018, 119(8): 6986-6996.
17. 马智勇, 郭成龙. lncRNA 对骨质疏松骨代谢相关信号通路影响的研究进展. 中国骨质疏松杂志, 2018, 24(4): 547-551.
18. Zhang TH, Liang LZ, Liu XL, et al. Long non-coding RNA MALAT1 interacts with miR-124 and modulates tongue cancer growth by targeting JAG1. Oncol Rep, 2017, 37(4): 2087-2094.
19. 田允鸿, 曾星, 邱慧芝, 等. MiR-124 对鼻咽癌细胞株上皮间充质转化和放疗敏感性的影响. 实用医学杂志, 2016, 32(17): 2780-2783.
20. 郭晓宇, 李唯, 陈克明, 等. 淫羊藿苷通过 PI3K/AKT-eNOS 信号途径促进大鼠骨髓基质细胞的成骨性分化. 中国药理学通报, 2013, 29(7): 966-970.

Chinese Journal of Reparative and Reconstructive Surgery

Study on adsorption of microRNA-124 by long chain non-coding RNA MALAT1 regulates osteogenic differentiation of mesenchymal stem cells

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