论文部分内容阅读
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。
Objective: To investigate the regulation of Runx2 and Osterix (OSX) overexpression on osteogenic differentiation of human umbilical vein endothelial cells (HUVECs). METHODS: Runx2 and Osterix genes were transfected into HUVECs by lentiviral vectors respectively. The effect of Runx2 and Osterix on osteogenic differentiation of HUVECs was investigated by alkaline phosphatase (ALP) staining and semi-quantitative assay. The expressions of Runx2, OSX, ALP, bone sialoprotein (BSP), osteopontin (OPN) and osteocalcin (OCN) in HUVECs were detected by RT-PCR, Western blotting and immunofluorescence staining. Data were statistically analyzed using the Graph Pad Prism 6.01 software package. RESULTS: Overexpression of Runx2 was beneficial to the osteogenic differentiation of HUVECs, whereas overexpression of Osterix did not. The transcript levels of Runx2, OSX, ALP, BSP, OPN and OCN were up-regulated in HUVECs transfected with Runx2 overexpression lentivirus, and the expressions of Runx2, OSX, OPN and OCN were also up-regulated. Conclusion: Overexpression of Runx2 can promote the osteogenic differentiation of HUVECs.