Runx2、Osterix转录因子过表达驱动内皮细胞成骨分化的机制探讨

doi:10.19439/j.sjos.2017.04.001

上海口腔医学 ›› 2017, Vol. 26 ›› Issue (4): 353-357.doi: 10.19439/j.sjos.2017.04.001

Runx2、Osterix转录因子过表达驱动内皮细胞成骨分化的机制探讨

杨光正¹, 张文杰², 丁迅², 张翔凯¹, 蒋欣泉^{2, *}, 张志愿^{1, *}

1.上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面-头颈肿瘤科,
2.口腔修复科,上海市口腔医学重点实验室,上海市口腔医学研究所,国家口腔疾病临床研究中心,上海 200011

收稿日期:2017-02-04 修回日期:2017-03-15 出版日期:2017-08-25 发布日期:2017-09-01
通讯作者: 张志愿,E-mail：zhzhy0502@163.com;
作者简介:杨光正（1990-）,男,硕士,E-mail：ygzwork@163.com
基金资助:
国家自然科学基金（81430012）

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

YANG Guang-zheng¹, ZHANG Wen-jie², DING Xun², ZHANG Xiang-kai¹, JIANG Xin-quan², ZHANG Zhi-yuan¹

1.Department of Oromaxillofacial Head and Neck Oncology,
2.Department of Prosthodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology; National Clinical Research Center of Stomatology. Shanghai 200011, China

Received:2017-02-04 Revised:2017-03-15 Online:2017-08-25 Published:2017-09-01

摘要/Abstract

摘要： 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GraphPad 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。

关键词: Runx2基因, Osterix基因, 成骨分化, 人脐静脉内皮细胞

Abstract: 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 immunofluorescence 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 osteocalcin (OCN), and up-regulated protein level of OPN and OCN. CONCLUTIONS: Overexpression of Runx2 could promote osteogenic differentiation of HUVECs.

Key words: Runx2 gene, Osterix gene, Osteogenic differentiation, Human umbilical vein endothelial cells

中图分类号:

R739.8

杨光正, 张文杰, 丁迅, 张翔凯, 蒋欣泉, 张志愿. Runx2、Osterix转录因子过表达驱动内皮细胞成骨分化的机制探讨[J]. 上海口腔医学, 2017, 26(4): 353-357.

YANG Guang-zheng, ZHANG Wen-jie, DING Xun, ZHANG Xiang-kai, JIANG Xin-quan, ZHANG Zhi-yuan. Effect of overexpression of transcription factor Runx2 and Osterix on osteogenic differentiation of endothelial cells[J]. Shanghai Journal of Stomatology, 2017, 26(4): 353-357.

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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

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