Runx2、Osterix转录因子过表达驱动内皮细胞成骨分化的机制探讨

Effect of overexpression of transcription factor Runx2 and Osterix on osteogenic differentiation of endothelial cells

目的:探讨Runx2和Osterix(OSX)过表达对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)成骨分化的调控作用。方法:通过慢病毒载体,将Runx2和Osterix基因分别转染入HUVECs。通过碱性磷酸酶(ALP)染色、半定量活性检测,探讨过表达Runx2和Osterix对HUVECs成骨分化的影响。通过RT-PCR、蛋白免疫印迹、免疫荧光染色检测成骨相关标志物Runx2、OSX、ALP、骨涎蛋白(BSP)、骨桥蛋白(OPN)、骨钙蛋白(OCN)在HUVECs中的表达。采用Graph Pad Prism 6.01软件包对数据进行统计学分析。结果:Runx2过表达有利于HUVECs的成骨分化,而Osterix过表达则无此作用。HUVECs转染Runx2过表达慢病毒后,成骨相关基因Runx2、OSX、ALP、BSP、OPN及OCN的转录水平上调,同时Runx2、OSX、OPN及OCN的蛋白表达水平亦有所上调。结论:Runx2过表达可促进HUVECs的成骨分化。

PURPOSE： To explore the effect of overexpression of Runx2 and Osterix （OSX） genes on osteogenic differentiation of human umbilical vein endothelial cells （HUVECs）. METHODS： Overexpressed Runx2 and OSX lentiviral vectors were transfected into HUVECs respectively. The osteogenic potential of transfected cells was identified by alkaline phosphatase （ALP） staining and ALP activity. Furthermore, real time-PCR, Western blot and immunofluorescenee staining were performed to detect the expression of osteogenic genes and proteins in HUVECs. GraphPad Prism 6.01 software was used for statistical analysis. RESULTS： Overexpression of Runx2 gene was beneficial for osteogenic differentiation of HUVECs, while overexpression of osterix gene did not show osteogenic differential potential. Moreover, overexpression of Runx2 gene in HUVECs up-regulated the gene expression level of Runx2, OSX, ALP, bone sialoprotein （BSP）, osteopontin （OPN）, and osteoealcin （OCN）, and up-regulated protein level of OPN and OCN. CONCLUTIONS： Overexpression of Runx2 could promote osteogenie differentiation of HUVECs.