miR-338-3p通过靶向RUNX2负调控人骨髓间充质干细胞成骨分化且促进骨质疏松  

miRNA-338-3p negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells and promotes the progression of osteoporosis by targeting RUNX2

作　　者：刘国樑[1] 秦明照[1] LIU Guo-liang;QIN Ming-zhao(Department of Geriatrics,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China)

机构地区：[1]首都医科大学附属北京同仁医院干部医疗/老年医学科,北京100730

出　　处：《川北医学院学报》2021年第5期551-556,共6页Journal of North Sichuan Medical College

基　　金：北京市保健委员会科研项目(京17-9)。

摘　　要：目的:探讨miRNA-338-3p在骨质疏松症进展中的作用及其对人骨髓间充质干细胞(hBMSCs)成骨分化的影响。方法:选择60例骨质疏松症患者为研究对象,另选同期60名非骨质疏松者作为对照组。检测两组血清miR-338-3p表达水平。采用RT-qPCR检测miR-338-3p、骨钙素(OCN)、骨桥蛋白(OPN)和I型胶原蛋白(collagen I)水平。Western blot检测miR-338-3p对RUNX2蛋白表达的影响;采用碱性磷酸酶(ALP)测定法和茜素红染色评估miR-338-3p和RUNX2对成骨分化和矿化能力的影响;采用生物信息学和荧光素酶报告基因检测验证miR-338-3p和RUNX2之间的关系。结果:骨质疏松患者血清miR-338-3p水平相比对照组表达量明显上升(P<0.05),且其表达随着成骨诱导时间的延长而逐渐降低(P<0.05)。与对照组比较,miR-338-3p表达情况影响OCN、OPN和Collagen I mRNA的水平(P<0.05)。与对照组相比,miR-338-3p过表达削弱了hBMSCs矿化能力和ALP活性(P<0.05)。双荧光素酶报告基因检测证实,RUNX2是miR-338-3p的直接靶点,miR-338-3p过表达降低了RUNX2蛋白及其mRNA的表达水平(P<0.05)。RUNX2过表达可逆转miR-338-3p对hBMSCs成骨分化的抑制作用。结论:miR-338-3p通过靶向RUNX2负调控hBMSCs的成骨分化,从而促进骨质疏松的进展。Objective:To investigate the role of miRNA-338-3p in the progression of osteoporosis and its effect on osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs).Methods:60 patients with osteoporosis were selected as the study subjects,and another 60 patients without osteoporosis were selected as the control group.The expression level of miR-338-3p in serum of the two groups was detected.Levels of miR-338-3p,osteocalcin(OCN),osteopontin(OPN)and Collagen I were detected by RT-qPCR.The effect of miR-338-3p on RUNX2 protein expression was detected by Western blot.The effects of miR-338-3p and RUNX2 on osteogenic differentiation and mineralization were evaluated by alkaline phosphatase(ALP)assay and alizarin red staining.Bioinformatics and luciferase reporter assay were used to verify the functional relationship between miR-338-3p and RUNX2.Results:Compared with the control group,the expression level of miR-338-3p in the serum of patients with osteoporosis was significantly increased(P<0.05),and it gradually decreased with the extension of osteogenic induction time(P<0.05).Compared with the control group,the expression of miR-338-3p obviously affected the levels of OCN,OPN and Collagen I mRNA(P<0.05).Compared with the control group,the overexpression of miR-338-3p notably impaired the mineralization ability and ALP activity of hBMSCs(P<0.05).Double luciferase reporter assay confirmed that RUNX2 was a direct target of miR-338-3p.Overexpression of miR-338-3p significantly reduced the mRNA and protein expression levels of RUNX2(P<0.05).RUNX2 overexpression reversed the inhibition of miR-338-3p on osteogenic differentiation of hBMSCs.Conclusion:miR-338-3p negatively regulated osteogenic differentiation of hBMSCs by targeting RUNX2,thus promoting the development of osteoporosis.

关 键 词：骨质疏松 成骨分化 人骨髓间充质干细胞成骨分化 MIRNA 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]