LncRNA ANRIL regulates cell proliferation and migration via sponging miR-339-5p and regulating FRS2 expression in atherosclerosis
LncRNA ANRIL regulates cell proliferation and migration via sponging miR-339-5p and regulating FRS2 expression in atherosclerosis
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LncRNA ANRIL 通过海绵 miR-339-5p 和调节动脉粥样硬化中的 FRS2 表达来调节细胞增殖和迁移
DOI：
10.26355/eurrev_202002_20373
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发表时间：
2020-02-01
影响因子：
3.3
通讯作者：
Yang, H-F
中科院分区：
文献类型：
--
作者：
Huang, T.;Zhao, H-Y;Yang, H-F
目的：动脉粥样硬化（AS）是世界范围内最常见的死亡原因之一，具有卒中或脑血管疾病的高风险。越来越多的证据表明INK 4基因座的长链非编码RNA（lncRNA）反义非编码RNA（ANRIL）与动脉粥样硬化血栓形成的易感性有关，并有助于AS的进展。患者和方法：人主动脉血管平滑肌细胞（HA-VSMCs）和人脐静脉内皮细胞（HU-VECs）与氧化低密度脂蛋白（ox-LDL）处理，并考虑作为AS细胞模型。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法分别检测mRNA和蛋白质表达水平。通过荧光原位杂交（FISH）的显微镜检查用于确定ANRIL的位置。细胞增殖和迁移实验证明ANRIL在AS中的功能作用。结果：在人动脉粥样硬化斑块和ox-LDL诱导的细胞中，ANRIL表达上调，miR-399- 5 p表达下调。ANRIL定位于细胞质，通过海绵状作用于miR-399- 5 p促进细胞增殖和迁移。进一步的分析确定成纤维细胞生长因子受体底物2（FRS 2）为miR-399- 5 p的直接靶点。结论：ANRIL通过吞噬miR-399- 5 p和调节RAS/RAF/ERK信号通路促进AS的发生发展，提示ANRIL可能是AS治疗的潜在靶点。
OBJECTIVE: Atherosclerosis (AS), with high risk of stroke or cerebrovascular disease, is one of the most common causes of death worldwide. Increasing evidence shows that long non-coding RNA (lncRNA) antisense non-coding RNA in the INK4 locus (ANRIL) is related to atherothrombotic stroke susceptibility and contributes to AS progression. However, the underlying mechanism was not explained yet.PATIENTS AND METHODS: Human aorta vascular smooth muscle cells (HA-VSMCs) and human umbilical vein endothelial cells (HU-VECs) were treated with oxidized Low Density Lipoprotein (ox-LDL) and considered as AS cell models. Quantitative reverse transcriptase PCR (qRT-PCR) and Western blotting were employed to investigate the mRNA and protein expression level, respectively. Microscopic examination through fluorescent in situ hybridization (FISH) was used to determine the location of ANRIL. Cell proliferation and migration assays were demonstrated to evaluate the functional role of ANRIL in AS. Potential target of ANRIL was determined using Luciferase reporter assay and RNA immunoprecipitation (RIP).RESULTS: ANRIL was upregulated and miR-399-5p was down-regulated in both human atherosclerotic plaques and ox-LDL-induced cells. ANRIL was located in cytoplasm and promoted cell proliferation and migration by sponging miR-399-5p. Further analysis identified fibroblast growth factor receptor substrate 2 (FRS2) as a direct target of miR-399-5p. Finally, RAS/RAF/ERK signal pathway was proved to be involved in the regulation of ANRIL on the progression of AS.CONCLUSIONS: These results revealed the underlying mechanism that ANRIL promoted AS progression by sponging miR-399-5p and regulating RAS/RAF/ERK signal pathway, suggesting that ANRIL might be a potential target for the therapeutic strategy of AS.