The Lnc-RNA APPAT Suppresses Human Aortic Smooth Muscle Cell Proliferation and Migration by Interacting With MiR-647 and FGF5 in Atherosclerosis
The Lnc-RNA APPAT Suppresses Human Aortic Smooth Muscle Cell Proliferation and Migration by Interacting With MiR-647 and FGF5 in Atherosclerosis
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Lnc-RNA APPAT 通过与动脉粥样硬化中的 MiR-647 和 FGF5 相互作用来抑制人主动脉平滑肌细胞的增殖和迁移
DOI：
10.1177/15266028221112247
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发表时间：
2022-07
期刊：
影响因子：
--
通讯作者：
Junwen Liu
中科院分区：
文献类型：
--
作者：
Fanming Meng;Luyang Han;Qin Liang;Shanshan Lu;Yanqing Huang;Junwen Liu
可以在循环血液中检测到LNCRNA-牵牛性斑块发病机理相关的转录本（APPAT），并已被证明与我们先前工作中的动脉粥样硬化的发展相关。它可能是早期诊断临床心血管疾病的潜在无创生物标志物。此外，在经OXLDL处理的血管平滑肌细胞和冠状动脉疾病患者的外周血中，miR-647的表达增加。确认了APPAT和miR-647之间的负相关性，并将FGF5筛选为miR-647的分子靶标。但是，目前尚不清楚APPAT，miR-647和FGF5如何在疾病发展中相互作用和功能。在这里，我们旨在通过定量逆转录聚合酶链反应检测到APPAT，miR-647和FGF5表达水平。通过EDU掺入分析检测到细胞增殖；通过伤口愈合测定检测细胞迁移；通过双 - 酸性酶报告基因测定法测定了APPAT/FGF5与miR-647的分子相互作用。进行蛋白质印迹以确定蛋白质水平的基因表达。通过原位杂交观察到APPAT和miR-647的亚细胞位置。进行了胞质和核分馏测定法进一步检测miR-647.APPAT和miR-647的分布对人主动脉平滑肌细胞（HASMCS）增殖和迁移具有反反应。 APTAT对细胞活性进行了负调节，而miR-647则以积极的方式进行（p <0.05）。发现了三对分子相互作用：APPAT和miR-647之间的相互负调节，APPAT下调FGF5，MiR-647 FGF5对FGF5的调节（P <0.05）。亚细胞位置测定证实了APPAT和miR-647的分子相互作用。APPAT可以通过与miR-647和FGF5相互作用来抑制OX-LDL处理的HASMC的迁移和增殖。我们揭示了在动脉粥样硬化进展中长期非编码RNA的非典型竞争性内源性RNA机制。
LncRNA-Atherosclerotic plaque pathogenesis-associated transcript (APPAT) could be detected in circulating blood and has been demonstrated to correlate with the development of atherosclerosis in our previous work. It could be a potential noninvasive biomarker for earlier diagnoses of clinical cardiovascular disease. Moreover, the expression of miR-647 increased in ox-LDL-treated vascular smooth muscle cells and peripheral blood of patients with coronary heart disease. A negative correlation between APPAT and miR-647 was confirmed, and FGF5 was screened as molecular target of miR-647. However, it is largely unclear how APPAT, miR-647, and FGF5 interact and function in disease development. Here, we aim to explore the underlying molecular mechanism in this progression.APPAT, miR-647, and FGF5 expression levels were detected by quantitative reverse transcription polymerase chain reaction; cell proliferation was detected by EdU incorporation assay; cell migration was detected by wound-healing assay; the molecular interaction of APPAT/FGF5 with miR-647 was verified by dual-luciferase reporter assay; the western blot was performed to determine the gene expression at protein levels; subcellular localizations of APPAT and miR-647 were observed by fluorescence in situ hybridization; cytosolic and nucleus fractionation assay was performed to further detect the distribution of miR-647.APPAT and miR-647 have inverse effects on human aortic smooth muscle cells' (HASMCs) proliferation and migration. APPAT negatively regulated the cell activity, whereas miR-647 did it in a positive way (p<0.05). Three pairs of molecular interplay were found: mutual negative regulation between APPAT and miR-647, APPAT downregulated FGF5, miR-647 regulation on FGF5 (p<0.05). Subcellular location assay confirmed the molecular interaction of APPAT and miR-647.APPAT could suppress the migration and proliferation of ox-LDL-treated HASMCs via interacting with miR-647 and FGF5. We revealed a nontypical competing endogenous RNA mechanism of long noncoding RNA in the progression of atherosclerosis.
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影响因子：
6.2
作者：
Majid, Shahana;Dar, Altaf A.;Saini, Sharanjot;Yamamura, Soichiro;Hirata, Hiroshi;Tanaka, Yuichiro;Deng, Guoren;Dahiya, Rajvir
通讯作者：
Dahiya, Rajvir
影响因子：
20.1
作者：
Kobiyama K;Ley K
通讯作者：
Ley K
DOI：
10.1161/atvbaha.114.303240
发表时间：
2014-06
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Bell RD;Long X;Lin M;Bergmann JH;Nanda V;Cowan SL;Zhou Q;Han Y;Spector DL;Zheng D;Miano JM
通讯作者：
Miano JM
影响因子：
14.9
作者：
Huang V;Place RF;Portnoy V;Wang J;Qi Z;Jia Z;Yu A;Shuman M;Yu J;Li LC
通讯作者：
Li LC
长链非编码RNA-RNCR3在动脉粥样硬化相关血管功能障碍中的作用
影响因子：
9
作者：
Shan K;Jiang Q;Wang XQ;Wang YN;Yang H;Yao MD;Liu C;Li XM;Yao J;Liu B;Zhang YY;J Y;Yan B
通讯作者：
Yan B