Histone modifications interact with DNA methylation at the GATA4 promoter during differentiation of mesenchymal stem cells into cardiomyocyte-like cells
Histone modifications interact with DNA methylation at the GATA4 promoter during differentiation of mesenchymal stem cells into cardiomyocyte-like cells
复制标题
间充质干细胞分化为心肌细胞样细胞期间，组蛋白修饰与 GATA4 启动子处的 DNA 甲基化相互作用
DOI：
10.1111/cpr.12253
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发表时间：
2016-06-01
影响因子：
8.5
通讯作者：
Zhu, Jing
中科院分区：
文献类型：
--
作者：
Xu, Hao;Yi, Qin;Zhu, Jing
目的我们前期的研究证实Islet-1特异性诱导MSCs向心肌细胞分化，其机制之一是调控组蛋白乙酰化。在此，我们从表观遗传修饰之间的相互作用的角度进一步探讨MSC分化为心肌细胞的机制。材料与方法我们使用慢病毒载体在MSC中过表达Islet-1，并进行ChIP-qPCR、MSP和BSP检测GATA 4和Nkx 2. 5启动子中组蛋白乙酰化/甲基化和DNA甲基化水平。为了进一步探讨这些表观遗传修饰之间的关系，我们使用5-aza或TSA干扰DNA甲基化和组蛋白乙酰化，分别，和检测对其他两个modification.ResultsHistone乙酰化水平增加，其甲基化水平下降，在GATA 4和Nkx2.5启动子; DNA甲基化水平降低，但在Nkx2.5启动子没有改变。此外，5-aza可增加组蛋白乙酰化水平，降低GATA 4启动子甲基化水平，但对Nkx2.5启动子无影响; TSA可降低GATA 4启动子区组蛋白甲基化和DNA甲基化水平，但仅降低Nkx2.5启动子区组蛋白甲基化水平。而Nkx2.5的表达不受DNA甲基化的调控。这三个修饰在GATA 4的调控过程中彼此高度相关，并在GATA 4启动子处产生了一个调控环。
ObjectivesA previous study of ours confirmed that Islet-1 specifically induces differentiation of MSCs into cardiomyocytes, and that one of the mechanisms underlying that process is regulation of histone acetylation. Here, we further explore the mechanism of MSC differentiation into cardiomyocytes from the perspective of interactions between epigenetic modifications.Materials and methodsWe used lentiviral vectors to overexpress Islet-1 in MSCs, and ChIP-qPCR, MSP and BSP were performed to detect levels of histone acetylation/methylation and DNA methylation in the GATA4 and Nkx2.5 promoters. To further explore relationships between these epigenetic modifications, we used 5-aza or TSA to interfere with DNA methylation and histone acetylation, respectively, and detected effects on the other two modifications.ResultsHistone acetylation level increased and its methylation level decreased at GATA4 and Nkx2.5 promoters; DNA methylation level was reduced at the GATA4 promoter but did not change at the Nkx2.5 promoter. Furthermore, 5-aza increased histone acetylation level and reduced its methylation level at the GATA4 promoter but had no effect on the Nkx2.5 promoter; TSA reduced histone methylation and DNA methylation levels at the GATA4 promoter, but it only reduced histone methylation level at the Nkx2.5 promoter.ConclusionsHistone acetylation/methylation and DNA methylation were both involved in regulating GATA4 expression, but Nkx2.5 expression was not regulated by DNA methylation. These three modifications had high correlation with each other during regulation of GATA4 and produced a regulation loop at the GATA4 promoter.