microRNA-1297 promotes the progression of osteoporosis through regulation of osteogenesis of bone marrow mesenchymal stem cells by targeting WNT5A
microRNA-1297 promotes the progression of osteoporosis through regulation of osteogenesis of bone marrow mesenchymal stem cells by targeting WNT5A
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DOI：
10.26355/eurrev_201906_18029
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发表时间：
2019-06-01
影响因子：
3.3
通讯作者：
Meng, Y.
中科院分区：
文献类型：
--
作者：
Wang, Q.;Wang, C-H;Meng, Y.
目的：探讨microRNA-1297是否通过Wnt5A调控骨髓间充质干细胞(BMSCs)的成骨，从而影响骨质疏松的进展。方法：采用实时定量聚合酶链式反应(qRT-PCR)方法检测骨质疏松患者和对照组的microRNA-1297水平及成骨相关标志物。用Western印迹法检测上述标志物和Wnt5A的蛋白水平。用碱性磷酸酶(ALP)活性测定和碱性磷酸酶染色检测在microRNA-1297和Wnt5A调控下的成骨分化程度，用ARS染色检测microRNA-1297过表达后hBMSC的矿化能力。用双荧光素酶报告实验确定了microRNA-1297与Wnt5A的结合位点。结果：MicroRNA-1297在骨质疏松症患者中高表达，且随着成骨诱导次数的增加，其表达水平显著降低。生物信息学预测表明，microRNA-1297可以靶向Wnt5A。体外实验表明，在hBMSC中过表达microRNA-1297可以降低Wnt5A的水平，而干扰microRNA-1297可以增加Wnt5A的水平。MicroRNA-1297的过表达和si-Wnt5A的转染显著降低了RUNX2、OSX、ALP、OCN、OPN和COL1A1的mRNA水平，从而抑制了成骨分化。结论：microRNA-1297可通过与Wnt5A结合，调节骨髓间充质干细胞的成骨分化，从而加速骨质疏松的进展。
OBJECTIVE: This study was designed to investigate whether microRNA-1297 can regulate the osteogenesis of bone marrow mesenchymal stem cells (BMSCs) through WNT5A, thus influencing the progression of osteoporosis.PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) assay was performed to analyze microRNA-1297 level and osteogenesis-related markers in osteoporosis patients and controls. The protein levels of the above markers and WNT5A were detected by Western blot. Alkaline phosphatase (ALP) activity assay and ALP staining were used to measure the degree of osteogenic differentiation under the control of microRNA-1297 and WNT5A, and ARS staining was used to detect the mineralization ability of hBMSC after overexpression of microRNA-1297. The binding sites of microRNA-1297 and WNT5A were determined by the dual luciferase-reporting assay. Besides, the activity of Wnt signal transduction pathway in different treatment groups was detected by TOP/FOP report.RESULTS: MicroRNA-1297 was highly expressed in osteoporotic patients, and its level decreased significantly with the increasing of osteogenic induction. Bioinformatics prediction suggested that microRNA-1297 can target WNT5A. In vitro experiments showed that overexpression of microRNA-1297 in hBMSC can reduce the level of WNT5A, while interference with microRNA-1297 can increase the level of WNT5A. Overexpression of microRNA- 1297 and transfection of si-WNT5A significantly reduced the mRNA levels of RUNX2, OSX, ALP, OCN, OPN and COL1A1, thereby inhibiting osteogenic differentiation. Overexpression of microRNA- 1297 could interfere with WNT signaling pathway regulation and regulate the osteogenic differentiation of hBMSCs.CONCLUSIONS: microRNA-1297 could regulate the osteogenesis of BMSCs by combining with WNT5A so as to accelerate the progression of osteoporosis.