抑制Runx2的表达增强BMP2诱导的干细胞成软骨分化  

Suppression of Runx2 Potentiates BMP2-induced Chondrogenic Differentiation

作　　者：孙泽绪 赵辰[1] 廖军义 王琦[1] 徐伟[1] 陈诚[1] 黄伟[1] 

机构地区：[1]重庆医科大学附属第一医院,重庆400016

出　　处：《中国生物工程杂志》2016年第4期57-62,共6页China Biotechnology

基　　金：国家自然科学基金面上项目(81371972)资助项目

摘　　要：目的:利用腺病毒介导RNA干扰抑制成骨分化关键转录调控因子Runx2(Runt-related transcription factor 2)的表达,研究其对骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)诱导间充质干细胞(mesenchymal stem cells,MSCs)成软骨分化的影响,并初步探讨相关机制。方法:利用腺病毒Ad-GFP、Ad-BMP2、Ad-SiRunx2感染C3H10T1/2细胞;共分4组:GFP组、BMP2组、BMP2+SiRunx2组和SiRunx2组。Alcian blue染色检测各组软骨细胞基质糖胺聚糖分泌;RTPCR检测Runx2、早期成软骨标志物(Col2a1)、蛋白聚糖(Aggrecan)及晚期成软骨标志物(Col10a1)mRNA表达水平;Western blot检测目的蛋白BMP2、Ⅱ型胶原(Col2a1)及X型胶原(Col10a1)的蛋白表达。结果:在BMP2诱导C3H10T1/2细胞成软骨分化过程中,下调Runx2的表达可以增强Col2a1、Aggrecan及软骨细胞外基质糖胺聚糖的合成,抑制Col10a1的合成。结论:下调Runx2表达可以增强BMP2诱导间充质干细胞成软骨分化能力,并抑制软骨细胞的成熟和肥大。Objective：To investigate the potentiation effect of RNA silenced Runx2 on bone morphogenetic protein 2（BMP2）induced chondrogenic differentiation of mesenchymal stem cells（MSCs）.Method：C3H10T1/2 cells were treated with adenovirus：Ad-BMP2,Ad-SiRunx2 or Ad-GFP.The sulfated glycosaminoglycan of cellular matrix was detected by Alcian Blue Staining.The mRNA level of Runx2,Col2a1,aggrecan,and Col10a1 were detected by RT-PCR,and the protein level of BMP2,Col2a1,Col10a1 were detected by Western blot.Results：The BMP2 ＋ SiRunx2 group（C3H10T1/2 cells were treated with Ad BMP2 and Ad SiRunx2）was more blue than the BMP2 group on day 10 proven by Alcian blue staining.The BMP2 ＋SiRunx2 group showed a higher mRNA level of Col2a1 and aggrecan（P〈0.05）and a lower protein level of Col10a1（P〈0.05）than the BMP2 group on day 7 and 14.The BMP2 ＋ SiRunx2 group showed a higher protein level of Col2a1（P〈0.05）and a lower protein level of Col10a1（P〈0.05）than the BMP2 group on day 10.Conclusion：The siRNA targeting Runx2 promotes BMP2-induced stem cell chondrogenic differentiation and supresses the chondrocyte hypertrophy and maturation.

关 键 词：RUNX2 骨形态发生蛋白2 C3H10T1/2细胞 成软骨分化 

分 类 号：Q254[生物学—细胞生物学]