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miR-920 通过靶向 HOXA7 促进人骨髓间充质干细胞的成骨分化。

MiR-920 promotes osteogenic differentiation of human bone mesenchymal stem cells by targeting HOXA7.

机构信息

Department of Orthopaedics, The Third Hospital of HeBei Medical University, 139 Ziqiang Road, Shijiazhuang, 050051, China.

出版信息

J Orthop Surg Res. 2020 Jul 10;15(1):254. doi: 10.1186/s13018-020-01775-7.

Abstract

BACKGROUND

To explore the effect of miR-920 on osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) and the possible mechanism.

METHODS

Osteoporosis (OP) and healthy control bone tissues were collected, and the relative expression of miR-920 and HOXA7 was measured. hBMSCs were isolated and cultured in vitro. Alkaline phosphatase activity and miR-920 and HOXA7 relative expression were measured during osteogenic differentiation of hBMSCs. Then, bioinformatic analysis was performed to assess the potential mechanism of miR-920. MiR-920 mimic and inhibitor were introduced into hBMSCs by lipofection transfection and were used to investigate the effect of miR-920 on the osteogenic differentiation of hBMSCs. A dual luciferase reporter assay was used to identify whether the 3'UTR of HOXA7 mRNA was a direct target of miR-920. Western blotting was performed to assess whether miR-920 affected the MAPK signaling pathway.

RESULTS

We found that miR-920 was downregulated in OP patients compared with controls, while HOXA7 was upregulated, and miR-920 had a negative correlation with HOXA7 (r = - 0.859, P = 0.001). Moreover, miR-920 was increased during osteogenic differentiation of hBMSCs, while HOXA7 had the opposite tendency. Bioinformatic analysis revealed that there were a total of 207 target genes, and MAPK was a potential targeted signaling pathway. MiR-920 mimic significantly increased ALP activity, calcium deposition, osteoblastic protein expression (ALP and OSX), and p-p38 and p-JNK protein levels.

CONCLUSION

Overall, miR-920 promotes osteogenic differentiation of hBMSCs by targeting HOXA7 through the MAPK signaling pathway.

摘要

背景

探讨 miR-920 对人骨髓间充质干细胞(hBMSCs)成骨分化的影响及其可能的机制。

方法

收集骨质疏松症(OP)和健康对照骨组织,测量 miR-920 和 HOXA7 的相对表达。体外分离培养 hBMSCs,检测 hBMSCs 成骨分化过程中碱性磷酸酶活性及 miR-920 和 HOXA7 的相对表达。然后,进行生物信息学分析以评估 miR-920 的潜在机制。通过脂质体转染将 miR-920 模拟物和抑制剂导入 hBMSCs,研究 miR-920 对 hBMSCs 成骨分化的影响。采用双荧光素酶报告基因检测鉴定 HOXA7 mRNA 的 3'UTR 是否为 miR-920 的直接靶标。采用 Western blot 检测 miR-920 是否影响 MAPK 信号通路。

结果

我们发现与对照组相比,OP 患者 miR-920 表达下调,HOXA7 表达上调,miR-920 与 HOXA7 呈负相关(r=-0.859,P=0.001)。此外,在 hBMSCs 成骨分化过程中 miR-920 增加,而 HOXA7 呈相反趋势。生物信息学分析显示共有 207 个靶基因,MAPK 是潜在的靶向信号通路。miR-920 模拟物显著增加碱性磷酸酶活性、钙沉积、成骨蛋白表达(碱性磷酸酶和 OSX)以及 p-p38 和 p-JNK 蛋白水平。

结论

综上所述,miR-920 通过靶向 HOXA7 并通过 MAPK 信号通路促进 hBMSCs 的成骨分化。

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