Transcription factor AP-2 gamma/Kruppel-like factor 10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension
Transcription factor AP-2 gamma/Kruppel-like factor 10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension
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DOI：
10.1097/hjh.0000000000003359
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发表时间：
2023-04-01
影响因子：
4.9
通讯作者：
Liu，Yawen
中科院分区：
文献类型：
--
作者：
Ren，Yaxuan;Shi，Jikang;Liu，Yawen
背景：内皮细胞功能障碍与微血管稀疏有关，反映了高血压的发病机制。我们之前的研究发现，miR-3656可以减少一氧化氮的生成和血管性血液病因子（vWF）的分裂，从而延缓血流并可能升高血压。在本文中，我们研究了在高血压中mir -3656损伤内皮细胞的转录调控机制。方法：分析miR-3656对内皮细胞增殖、迁移、成管和凋亡功能的影响。采用实时荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction, qRT-PCR）和western blot检测基因mRNA和蛋白水平。双荧光素酶报告基因检测证实miR-3656与转录因子AP-2 γ (TFAP2C)的3 '非翻译区（UTR）之间的结合。染色质免疫沉淀- qpcr法证实TFAP2C与kr<s:1> ppel样因子10 （KLF10）启动子区结合。结果：miR-3656对内皮细胞增殖、迁移、成管和凋亡均有影响。miR-3656通过直接靶向TFAP2C的3'UTR抑制TFAP2C的表达；此外，TFAP2C过表达可挽救mir -3656诱导的内皮细胞损伤。此外，TFAP2C下调通过结合KLF10启动子区域降低KLF10的表达，而KLF10上调逆转了TFAP2C沉默对内皮细胞的影响。这些抑制过程导致miR-3656干扰klf10促进内皮细胞的功能。结论：TFAP2C/KLF10轴参与高血压患者内皮细胞mir -3656相关功能障碍。TFAP2C和KLF10启动子区域的3'UTR是TFAP2C/KLF10轴的枢纽。
Background:Dysfunction of endothelial cells links to microvascular rarefaction, reflecting the pathogenesis of hypertension. Our previous studies found that miR-3656 reduces nitric oxide generation and von Willebrand factor (vWF) cleavage, thereby retarding blood flow and potentially increasing blood pressure. In this paper, we investigated mechanism of transcription regulation contributing to miR-3656-damaged endothelial cells in hypertension.Methods:The effects of miR-3656 on function of endothelial cells were analyzed on the basis of proliferation, migration, tube formation, and apoptosis. The mRNA level and protein level of genes were examined using quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Dual-luciferase reporter assay was performed to confirm the binding between miR-3656 and 3’untranslated region (UTR) of transcription factor AP-2 gamma (TFAP2C). The binding between TFAP2C and the promoter region of Krüppel-like factor 10 (KLF10) was confirmed by chromatin immunoprecipitation-qPCR assay.Results:miR-3656 impaired the cell proliferation, migration, tube formation, and apoptosis of endothelial cells. miR-3656 inhibited the expression of TFAP2C by directly targeting 3’UTR of TFAP2C; moreover, miR-3656-induced injury of endothelial cells was rescued by TFAP2C overexpression. Furthermore, downregulated TFAP2C decreased KLF10 expression by binding to KLF10 promoter region, and upregulated KLF10 reversed the effects of silencing TFAP2C on endothelial cells. These inhibitory processes led to interference of miR-3656 to KLF10-promoted function of endothelial cells.Conclusion:TFAP2C/KLF10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension. The 3’UTR of TFAP2C and KLF10 promoter region are the hubs of the TFAP2C/KLF10 axis.