Methylation status of CpG sites in the NOTCH4 promoter region regulates NOTCH4 expression in patients with tetralogy of Fallot.
Methylation status of CpG sites in the NOTCH4 promoter region regulates NOTCH4 expression in patients with tetralogy of Fallot.
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NOTCH4启动子区CpG位点的甲基化状态调节法洛四联症患者NOTCH4的表达
DOI：
10.3892/mmr.2020.11535
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发表时间：
2020-11
影响因子：
3.4
通讯作者：
Huang G
中科院分区：
文献类型：
--
作者：
Zhu Y;Ye M;Xu H;Gu R;Ma X;Chen M;Li X;Sheng W;Huang G
法洛四联症（TOF）是紫绀型先天性心脏病（CHD）最常见的形式。虽然TOF患者的全基因组甲基化水平较低，但对特定基因的DNA甲基化变化及其与TOF发生的关系知之甚少。NOTCH 4是Notch信号通路的介导物，在正常心脏发育中起重要作用。然而，NOTCH 4基因的表观遗传调控在TOF发病机制中的作用仍不清楚。考虑到NOTCH 4基因在TOF患者中突变频率低，表达降低，我们推测NOTCH 4基因的异常DNA甲基化改变可能影响其表达，从而导致TOF的发生。本研究通过检测NOTCH 4基因启动子区甲基化状态，探讨其可能与TOF疾病相关的调控机制。此外，为了进一步了解可能在TOF发展中起作用的表观遗传机制，测量了NOTCH 4的启动子甲基化状态。免疫组化法检测TOF患者右室流出道心肌组织中NOTCH 4的表达。与健康对照组相比，TOF患者的NOTCH 4表达显著降低（P=0.0055）。此外，亚硫酸氢盐测序表明，在NOTCH 4启动子中的CpG位点2的甲基化水平在患者中显著高于对照组（P=0.0459）。NOTCH 4表达与CpG位点2甲基化水平呈负相关（r=-0.51; P=0.01）。ETS 1转录因子可以通过与靶基因的特异性DNA序列结合而作为转录激活因子，如DLL 4和NOTCH 4，其在正常心脏发育中起重要作用。双荧光素酶报告和电泳迁移率变动分析表明，ETS 1转录因子可以结合到NOTCH 4启动子区。然而，ETS 1与NOTCH 4启动子的结合被假定的ETS 1结合位点的甲基化所废除。这些发现表明，TOF患者中NOTCH 4表达的降低可能与NOTCH 4启动子区域中CpG位点2的高甲基化有关，这是由于ETS 1结合受损所致。
Tetralogy of Fallot (TOF) is the most common form of cyanotic congenital heart disease (CHD). Although a lower methylation level of whole genome has been demonstrated in TOF patients, little is known regarding the DNA methylation changes in specific gene and its associations with TOF development. NOTCH4 is a mediator of the Notch signalling pathway that plays an important role in normal cardiac development. However, the role of epigenetic regulation of the NOTCH4 gene in the pathogenesis of TOF remains unclear. Considering the NOTCH4 low mutation frequency and reduced expression in the TOF patients, we hypothesized that abnormal DNA methylation change of NOTCH4 gene may influence its expression and responsible for TOF development. In this study, we measured the promoter methylation status of NOTCH4 and was measured and its regulation mechanism was explored, which may be related to TOF disease. Additionally, the promoter methylation statuses of NOTCH4 was measured in order to further understand epigenetic mechanisms that may serve a role in the development of TOF. Immunohistochemical analysis was used to examine NOTCH4 expression in right ventricular outflow tract myocardial tissues in patients with TOF. Compared with healthy controls, patients with TOF displayed significantly reduced in NOTCH4 expression (P=0.0055). Moreover, bisulphite sequencing suggested that the methylation levels of CpG site 2 in the NOTCH4 promoter was significantly higher in the patients than in the controls (P=0.0459). NOTCH4 expression was negatively associated with CpG site 2 methylation levels (r=−0.51; P=0.01). ETS1 transcription factor can serve as transcriptional activators by binding to specific DNA sequences of target genes, such as DLL4 and NOTCH4, which serves an important role in normal heart development. Dual-luciferase reporter and electrophoretic mobility shift assays indicated that the ETS1 transcription factor could bind to the NOTCH4 promoter region. However, binding of ETS1 to the NOTCH4 promoter was abrogated by methylation at the putative ETS1 binding sites. These findings suggested that decreased NOTCH4 expression in patients with TOF may be associated with hypermethylation of CpG site 2 in the NOTCH4 promoter region, due to impaired binding of ETS1.
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影响因子：
20.1
作者：
Kathiriya IS;Nora EP;Bruneau BG
通讯作者：
Bruneau BG
DOI：
10.1016/j.jsbmb.2012.01.009
发表时间：
2012-05-01
影响因子：
4.1
作者：
Fuerst, Rainer W.;Kliem, Heike;Ulbrich, Susanne E.
通讯作者：
Ulbrich, Susanne E.
影响因子：
4.6
作者：
Gao, Zhiguang;Kim, Gene H.;Svensson, Eric C.
通讯作者：
Svensson, Eric C.
影响因子：
5.3
作者：
Leong, KG;Hu, XL;Karsan, A
通讯作者：
Karsan, A
DOI：
10.1073/pnas.90.16.7588
发表时间：
1993-08-15
影响因子：
11.1
作者：
KOLA, I;BROOKES, S;SETH, A
通讯作者：
SETH, A