Department of Respiratory and Critical Care Medicine, Beijing Chaoyang Hospital affiliated to Capital Medical University, Beijing 100020, China.
Department of Respiratory and Critical Care Medicine, Beijing Chaoyang Hospital affiliated to Capital Medical University, Beijing 100020, China.
Life Sci. 2020 Aug 15;255:117835. doi: 10.1016/j.lfs.2020.117835. Epub 2020 May 23.
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.
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.
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.
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.
新出现的研究结果表明，非编码 RNA（ncRNA）在哮喘发病机制中起着关键作用。然而，环状 RNA（circRNA）在气道重塑中的生物学作用仍不清楚。本研究重点关注环状 RNA circHIPK3 在气道平滑肌细胞（ASMC）增殖和迁移中的调节作用。
使用 Sanger 测序检测 circRNA 的序列。使用 CCK-8 测定法、transwell 和流式细胞仪测定法检测细胞表型。使用双荧光素酶报告基因测定法检测 miRNA 和下游及上游靶标的潜在结合。
结果表明，血小板衍生生长因子（PDGF）诱导的 ASMC 中 circHIPK3 显著上调。使用 CCK-8、transwell 迁移测定法和流式细胞术分析的功能分析表明，circHIPK3 敲低抑制了 ASMC 的增殖、迁移并上调了细胞凋亡。机制分析表明，circHIPK3 在细胞质中海绵吸附 miR-326，从而靶向基质相互作用分子 1（STIM1）来调节 ASMC 的增殖、迁移和凋亡。
总之，这些数据表明，circHIPK3 通过 miR-326/STIM1 轴在哮喘发病过程中的气道重塑中发挥调节作用，为治疗靶点提供了新的见解。
