miR-183-96-182簇通过靶向FOXO1减轻氧化型低密度脂蛋白诱导的血管内皮细胞凋亡。
miR-183-96-182 clusters alleviated ox-LDL-induced vascular endothelial cell apoptosis by targeting FOXO1.
作者信息
Liu Zhi-Qin, Du Jing-Jing, Ren Jing-Jing, Zhang Zhi-Yong, Guo Xiao-Bo, Yan Yu-E, Jia Xiao-Tao, Gu Nai-Bing, Di Zheng-Li, Li San-Zhong
机构信息
Department of Neurology, Xi'an Central Hospital, Xi'an Jiaotong University, School of Medicine Xi'an 710003 Shaanxi China.
Department of Hematology, Xi'an Central Hospital, Xi'an Jiaotong University, School of Medicine Xi'an 710003 Shaanxi China.
出版信息
RSC Adv. 2018 Oct 12;8(61):35031-35041. doi: 10.1039/c8ra06866f. eCollection 2018 Oct 10.
OBJECTIVE
To investigate the role of FOXO1 and miR-183-96-182 clusters in ox-LDL induced endothelial cell apoptosis.
METHODS
FOXO1 overexpression (OE) and knockdown (KD) as well as AKT1 OE in human umbilical vein endothelial cells (HUVECs) and human aortic endothelial cells (HAECs) were achieved by lentiviral transduction. Upregulation of miR-183-5p, miR-182-5p or miR-96-5p was mimicked by agomir treatment. FOXO1 gene transcription was monitored by FOXO1 promotor reporter assay. Cell apoptosis in culture was monitored by TiterTACS detection. Regulation of FOXO1 gene expression by an miRNA targeting mechanism was monitored by AGO2-RNA immunoprecipitation assay.
RESULTS
FOXO1 mRNA and protein expression levels in ox-LDL treated HUVECs or HAECs were significantly upregulated due to transcriptional and miRNA targeting mechanisms. MiR-183-5p, miR-182-5p and miR-96-5p expression levels in HUVECs or HAECs were significantly reduced by ox-LDL treatment, the overexpression of which by agomir treatment partially reduced the FOXO1 mRNA/protein expression levels and cell apoptosis which was upregulated by ox-LDL treatment. FOXO1 overexpression antagonized the effect of the agomir treatment indicated above. MiR-183-5p, miR-182-5p and miR-96-5p agomir treatment partially rescued the FOXO1 pSer256/total FOXO1 protein ratio and the AKT1 pSer473 level that were reduced by ox-LDL treatment in the HUVECs or HAECs. AKT1 overexpression significantly reduced FOXO1 protein expression, increased miR-182-5p and miR-183-5p expression, and partially alleviated ox-LDL induced HUVEC or HAEC apoptosis in an miR-183-5p and miR-182-5p-dependent manner.
CONCLUSION
miR-183-96-182 clusters could partially alleviate ox-LDL-induced apoptosis in HUVECs or HAECs by targeting FOXO1.
目的
研究FOXO1和miR-183-96-182簇在氧化型低密度脂蛋白（ox-LDL）诱导的内皮细胞凋亡中的作用。
方法
通过慢病毒转导实现人脐静脉内皮细胞（HUVECs）和人主动脉内皮细胞（HAECs）中FOXO1的过表达（OE）和敲低（KD）以及AKT1的过表达。通过agomir处理模拟miR-183-5p、miR-182-5p或miR-96-5p的上调。通过FOXO1启动子报告基因检测监测FOXO1基因转录。通过TiterTACS检测监测培养中的细胞凋亡。通过AGO2-RNA免疫沉淀检测监测miRNA靶向机制对FOXO1基因表达的调控。
结果
由于转录和miRNA靶向机制，ox-LDL处理的HUVECs或HAECs中FOXO1的mRNA和蛋白表达水平显著上调。ox-LDL处理显著降低了HUVECs或HAECs中miR-183-5p、miR-182-5p和miR-96-5p的表达水平，通过agomir处理过表达这些miRNA可部分降低FOXO1的mRNA/蛋白表达水平以及ox-LDL处理上调的细胞凋亡。FOXO1过表达拮抗上述agomir处理的效果。miR-183-5p、miR-182-5p和miR-96-5p的agomir处理可部分挽救HUVECs或HAECs中被ox-LDL处理降低的FOXO1 pSer256/总FOXO1蛋白比值和AKT1 pSer473水平。AKT1过表达显著降低FOXO1蛋白表达，增加miR-182-5p和miR-183-5p表达，并以miR-183-5p和miR-182-5p依赖的方式部分减轻ox-LDL诱导的HUVEC或HAEC凋亡。
结论
miR-183-96-182簇可通过靶向FOXO1部分减轻ox-LDL诱导的HUVECs或HAECs凋亡。
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