Transcription factor AP-2 gamma/Krüppel-like factor 10 axis is involved in miR-3656-related dysfunction of endothelial cells in hypertension

Abstract

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 ofTFAP2Cby directly targeting 3’UTR ofTFAP2C; moreover, miR-3656-induced injury of endothelial cells was rescued byTFAP2Coverexpression. Furthermore, downregulated TFAP2C decreasedKLF10expression by binding toKLF10promoter region, and upregulatedKLF10reversed the effects of silencingTFAP2Con 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 ofTFAP2CandKLF10promoter region are the hubs of the TFAP2C/KLF10 axis.