Department of Orthopaedics, The Third Hospital of HeBei Medical University, 139 Ziqiang Road, Shijiazhuang, 050051, China.
J Orthop Surg Res. 2020 Jul 10;15(1):254. doi: 10.1186/s13018-020-01775-7.
To explore the effect of miR-920 on osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) and the possible mechanism.
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.
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.
Overall, miR-920 promotes osteogenic differentiation of hBMSCs by targeting HOXA7 through the MAPK signaling pathway.
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。
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。
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。
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。