miRNA-144靶向调节Cadherin-11对间充质干细胞成骨分化的抑制效应.doc

miRNA-144靶向调节Cadherin-11对间充质干细胞成骨分化的抑制效应 康 夏1,2，康 菲1，杨 波1，郭洪峰1，权 毅2，董世武1 （400038 重庆，第三军医大学解剖学教研室1；610083 成都，解放军成都军区总医院骨科2） [摘要] 目的 证实miRNA-144靶向调节钙粘蛋白11(Cad-11)表达，抑制间充质干细胞（mesenchymal stem cell，MSC）的成骨分化。 方法 通过双荧光素酶报告系统对Cad-11与miRNA-144的靶基因关系进行鉴定；将pre-miR-144及anti-miR-144分别转染MSC，成骨诱导7、14 d后，实时定量PCR和蛋白印迹方法检测细胞中Cad-11及成骨标志物Cbfa1/Runx2、ALP的表达情况，分析其与MSC成骨分化的变化关系。粘附实验检测miRNA-144对MSC成骨分化过程中粘附的影响。结果 Cad-11是miRNA-144的靶基因；miRNA-144在成骨诱导第7、14天可明显抑制Cad-11、Runx2和ALP的表达(P0.05)，同时降低MSCs细胞的粘附性。结论 miRNA-144通过抑制Cad-11的表达对间充质干细胞的成骨分化起负向调控的作用。 [关键词] miRNA-144；Cadherin-11；MSC；成骨分化 [中图法分类号] [文献标志码] A [基金项目] 国家重点基础研究发展计划项目（2011CB964701）；国家自然科学基金项目81271980） [通信作者] 权 毅，E-mail：quanyi56@;董世武，E-mail：dongshiwu@163.com miRNA-144 inhibiting the osteogenesis of MSCs by targeting Cad-11 Kang Xia, Kang Fei, Yang Bo, Guo Hongfeng, Quan Yi, Dong Shiwu (1Laboratory of Biomechanics, Department of Anatomy, The Third Military Medical University, Chongqing,400038; 2Department of Orthopaedics, Chengdu General Military Hospital, Chengdu, Sichuan Province,610083,China) [Abstract] Objective To investigate the role of miRNA-144 in inhibiting the osteogenesis of mesenchymal stem cells （MSCs）by regulating Cadherin-11（Cad-11）. Method To identify the the relationship between miRNA-144 and Cad-11 by using dual-luciferase reporter assay. Pre-mir-144 and anti-mir-144 were transfected into the C3H/10T1/2 cells, respectively. Then, the cells were collected after osteogenesis induced for 7 days and 14 days. Real-time PCR and Western Blotting assays were performed to detect the expression of Cad-11 and related osteogenetic biomarkers, including Runx2 and ALP. Then the cell to cell adhesion was tested. Results Cad-11 was the target gene of miRNA-144, and miRNA-144 can downregulate the expression of Cad-11,Runx2 and C22, furthermore, it can inhibit the adhesion of MSCs. Conclusion miRNA-144 can inhibit the osteogenesis process of MSCs by downregulating the Cad-11. [Key Words] miRNA-