EPHA2在人骨髓间充质干细胞成骨分化中的作用  

The Role of EPHA2 in Osteogenic Differentiation of Human Bone Marrow Mesenchymal Stem Cells

作　　者：孙博[1] 张弢[2] 杨龙[1] 李靖[1] 任厚相 孙琦[1] 王建吉[1] 叶川[1] 

机构地区：[1]贵阳医学院 [2]中国人民解放军第117医院骨科,浙江杭州311201

出　　处：《贵阳医学院学报》2015年第5期433-437,共5页Journal of Guiyang Medical College

基　　金：国家自然科学基金资助项目(81360232);贵州省卫生厅资助项目(gzwkj2010-1-042);贵州省国际科技合作项目(黔科合外G字2010-7);贵州省科技厅社发联合基金项目(黔科合20103166)

摘　　要：目的:研究EPHA2在人骨髓间充质干细胞(h BMSC)成骨分化过程中表达的动态变化及其在成骨分化中的作用。方法:h BMSC成骨分化诱导0、4、10和14 d后分别收集细胞提取RNA和蛋白,采用Real-time PCR和Western blot方法检测EPHA2 mRNA及蛋白表达;使用EPHA2的si-RNA下调EPHA2表达后进行成骨分化诱导,检测下调EPHA2表达对早期成骨分化指标ALP活性及晚期成骨分化指标钙沉积、成骨分化标志物OSX、OCN和成骨分化关键转录因子RUNX2表达的影响。结果:EPHA2在h BMSC成骨分化过程中表达逐渐上升,下调EPHA2表达能抑制ALP活性和钙沉积,抑制OSX、OCN及关键转录因子RUNX2的表达。结论:EPHA2在h BMSC成骨分化过程中表达逐渐上升,EPHA2可能通过增强RUNX2的表达从而促进h BMSC成骨分化。Objective： To investigate the dynamic changes of EPHA2 expression in the osteogenic differentiation of human bone marrow mesenehymal stem cells（ hBMSC） and the role of EPHA2 in the osteogenic differentiation. Methods： Cells were harvested on day 0,day 4,day 10 and day 14 during induction of osteogenic differentiation,RNA and protein was extracted and detected with Real-time PCR and Western blot analysis. siRNA for EPHA2 was used to silence the expression of EPHA2. Alkaline phosphatase（ ALP） activity,calcium deposition,and the levels of OSX,OCN,RUNX2 were detected to observe the effect of EPHA2 on the osteogenic differentiation of hBMSC. Results： The expression of EPHA2 increased gradually in hBMSC osteogenic differentiation process. Down-regulation of EPHA2 inhibited ALP activity,calcium deposition and the expression of OSX,OCN and RUNX2 in hBMSC. Conclusions： The level of EPHA2 increases gradually in hBMSC osteogenic differentiation process,and EPHA2 can promote hBMSC osteogenic differentiation process through enhancing the expression of RUNX2.

关 键 词：EPHA2 人骨髓间充质干细胞 成骨分化 RUNX2 

分 类 号：R318.17[医药卫生—生物医学工程]