Twist-related protein 1 negatively regulated osteoblastic transdifferentiation of human aortic valve interstitial cells by directly inhibiting runt-related transcription factor 2.
Twist-related protein 1 negatively regulated osteoblastic transdifferentiation of human aortic valve interstitial cells by directly inhibiting runt-related transcription factor 2.
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DOI：
10.1016/j.jtcvs.2014.02.084
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发表时间：
2014-10
期刊：
影响因子：
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通讯作者：
Xi-wu Zhang;Bo-yao Zhang;Shu-wei Wang;De-jun Gong;Lin Han;Zhiyun Xu;Xiao-hong Liu
中科院分区：
文献类型：
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作者：
Xi-wu Zhang;Bo-yao Zhang;Shu-wei Wang;De-jun Gong;Lin Han;Zhiyun Xu;Xiao-hong Liu
目的：瓣膜钙化涉及到瓣膜间质细胞向成骨细胞的转分化。twist相关蛋白1 （TWIST1）已被确定为小鼠和人间充质干细胞成骨细胞分化的负调节因子，但其在人主动脉vic中的功能尚不清楚。在我们的研究中，我们确定了TWIST1在调节人主动脉静脉成骨细胞转分化中的作用机制。方法对人钙化和非钙化主动脉瓣进行TWIST1表达检测。分离培养人主动脉vic。结果钙化主动脉瓣中TWIST1表达水平较低。体外实验表明，TWIST1过表达通过降低runt相关转录因子2 （RUNX2）及其下游成骨细胞标志物的表达，抑制了vic向成骨细胞的转分化。通过染色质免疫沉淀和双荧光素酶实验，我们发现TWIST1通过直接结合位于RUNX2 P2启动子区域−820 bp的E-box并抑制其活性来抑制RUNX2的表达。结论TWIST1可能通过直接抑制RUNX2负向调节人主动脉内皮细胞成骨转分化，从而起到预防人主动脉瓣钙化的重要作用。
ObjectiveValve calcification involves transdifferentiation of valve interstitial cells (VICs) into osteoblasts. Twist-related protein 1 (TWIST1) has been established as a negative regulator of osteoblast differentiation in both mouse and human mesenchymal stem cells, but its function in human aortic VICs is unknown. In our study, we determined the mechanism of TWIST1 action in regulating osteoblastic transdifferentiation of human aortic VICs.MethodsHuman calcified and noncalcified aortic valves were examined for TWIST1 expression. Human aortic VICs were isolated and cultured.ResultsThe data showed that calcified aortic valves express lower levels of TWIST1. In vitro experiments showed that TWIST1 overexpression inhibited the transdifferentiation of VICs into osteoblasts by decreasing the expression of runt-related transcription factor 2 (RUNX2) and its downstream osteoblastic markers. Through chromatin immunoprecipitation and dual luciferase assays, we found that TWIST1 repressed the expression of RUNX2 by directly binding to an E-box located at −820 bp of the RUNX2 P2 promoter region and inhibiting its activity.ConclusionsOur study results suggest that TWIST1 could play an important role in preventing human aortic valve calcification by negatively regulating osteoblastic transdifferentiation of human aortic VICs through direct inhibition of RUNX2.