环状RNA circHIPK3通过miR-326/STIMI轴调节气道平滑肌细胞的增值
2020/10/14
摘要
目的：新发现证明了非编码RNA（ncRNA）在哮喘发展中的关键作用。尽管如此，环状RNA（circRNA）在气道重塑中的生物学作用仍然难以捉摸。在此，目前的研究集中在circRNA circHIPK3在气道平滑肌细胞（ASMCs）增值和迁移中的调控。
材料和方法：使用Sanger测序检测circRNA的序列。采用CCK-8、transwell和流氏细胞仪检测细胞表型。使用双荧光素酶报告基因检测miRNA与下游和上游靶点的潜在结合。
关键发现：结果显示circRNAHIPK3在血小板衍生生长因子（PDGF）诱导的ASMCs中显著上调。使用CCK8、transwell迁移实验和流氏细胞术的功能分析表明，circHIPK3敲除抑制ASMCs的增殖、迁移和上调细胞凋亡。机制分析表明，在细胞质中circHIPK3吸收miR-326，从而靶向机制相互作用分子1（STIMI）调节ASMCs的增殖、迁移和凋亡。
意义：总的来说，结果阐明了circHIPK3在哮喘发展过程中通过miR326/STIMI轴作为气道重塑的调节因子，为治疗靶点提供了新的见解。
Circular RNA circHIPK3 modulates the proliferation of airway smooth muscle cells by miR-326/STIM1 axis
Junling Lin,Xiaokai Feng,Jun Zhang
Abstract
Aims: Emerging findings demonstrate the critical roles of noncoding RNA (ncRNA) in asthma development. Nevertheless, the biological roles of circular RNA (circRNA) in airway remodeling are still elusive. Here, the present research focuses on the regulation of circRNA circHIPK3 in airway smooth muscle cells (ASMCs) proliferation and migration.
Materials and methods: The sequence of circRNA was detected using Sanger sequencing. Cellular phenotypes were detected using CCK-8 assay, transwell and flow cytometer assay. The potential binding of miRNA and downstream and upstream targets was detected using dual-luciferase reporter assay.
Key findings: Results showed that circHIPK3 was significantly upregulated in platelet-derived growth factor (PDGF) induced ASMCs. Functional analysis using CCK-8, transwell migration assays and flow cytometry analysis showed that circHIPK3 knockdown repressed proliferation, migration and up-regulated the apoptosis in ASMCs. Mechanistic assays showed that circHIPK3 sponged miR-326 in the cytoplasm, thereby targeting stromal interaction molecule 1 (STIM1) to regulate ASMCs' proliferation, migration and apoptosis.
Significance: Collectively, the data elucidates that circHIPK3 functions as a regulator in the airway remodeling during the asthma development through miR-326/STIM1 axis, providing a novel insight for the therapeutic target.
目的：新发现证明了非编码RNA（ncRNA）在哮喘发展中的关键作用。尽管如此，环状RNA（circRNA）在气道重塑中的生物学作用仍然难以捉摸。在此，目前的研究集中在circRNA circHIPK3在气道平滑肌细胞（ASMCs）增值和迁移中的调控。
材料和方法：使用Sanger测序检测circRNA的序列。采用CCK-8、transwell和流氏细胞仪检测细胞表型。使用双荧光素酶报告基因检测miRNA与下游和上游靶点的潜在结合。
关键发现：结果显示circRNAHIPK3在血小板衍生生长因子（PDGF）诱导的ASMCs中显著上调。使用CCK8、transwell迁移实验和流氏细胞术的功能分析表明，circHIPK3敲除抑制ASMCs的增殖、迁移和上调细胞凋亡。机制分析表明，在细胞质中circHIPK3吸收miR-326，从而靶向机制相互作用分子1（STIMI）调节ASMCs的增殖、迁移和凋亡。
意义：总的来说，结果阐明了circHIPK3在哮喘发展过程中通过miR326/STIMI轴作为气道重塑的调节因子，为治疗靶点提供了新的见解。
(中日友好医院呼吸与危重症医学科 张清 摘译 林江涛 审校)
(Life Sci. 2020 Aug 15; 255:117835. doi: 10.1016/j.lfs.2020.117835. Epub 2020 May 23.)
(Life Sci. 2020 Aug 15; 255:117835. doi: 10.1016/j.lfs.2020.117835. Epub 2020 May 23.)
Circular RNA circHIPK3 modulates the proliferation of airway smooth muscle cells by miR-326/STIM1 axis
Junling Lin,Xiaokai Feng,Jun Zhang
Abstract
Aims: Emerging findings demonstrate the critical roles of noncoding RNA (ncRNA) in asthma development. Nevertheless, the biological roles of circular RNA (circRNA) in airway remodeling are still elusive. Here, the present research focuses on the regulation of circRNA circHIPK3 in airway smooth muscle cells (ASMCs) proliferation and migration.
Materials and methods: The sequence of circRNA was detected using Sanger sequencing. Cellular phenotypes were detected using CCK-8 assay, transwell and flow cytometer assay. The potential binding of miRNA and downstream and upstream targets was detected using dual-luciferase reporter assay.
Key findings: Results showed that circHIPK3 was significantly upregulated in platelet-derived growth factor (PDGF) induced ASMCs. Functional analysis using CCK-8, transwell migration assays and flow cytometry analysis showed that circHIPK3 knockdown repressed proliferation, migration and up-regulated the apoptosis in ASMCs. Mechanistic assays showed that circHIPK3 sponged miR-326 in the cytoplasm, thereby targeting stromal interaction molecule 1 (STIM1) to regulate ASMCs' proliferation, migration and apoptosis.
Significance: Collectively, the data elucidates that circHIPK3 functions as a regulator in the airway remodeling during the asthma development through miR-326/STIM1 axis, providing a novel insight for the therapeutic target.
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调节性B细胞控制鼠哮喘模型中气道高反应性和肺部重塑
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TSLP和IL-33相互促进肺蛋白表达和ILC2受体表达增强先天2型气道炎症
