Miao Ran, Dong Xingbei, Gong Juanni, Wang Ying, Guo Xiaojuan, Li Yidan, Liu Min, Wan Jun, Li Jifeng, Yang Suqiao, Wang Wang, Kuang Tuguang, Zhong Jiuchang, Zhai Zhenguo, Yang Yuanhua
Medical Research Center, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
Key Laboratory of Respiratory and Pulmonary Circulation Disorders, Institute of Respiratory Medicine, Beijing, China.
Pulm Circ. 2020 Jul 10;10(3):2045894020928300. doi: 10.1177/2045894020928300. eCollection 2020 Jul-Sep.
Chronic thromboembolic pulmonary hypertension (CTEPH) is characterized by elevated pressure in pulmonary arteries. This study was performed to explore the critical miRNAs and genes affecting the pathogenesis of CTEPH.
GSE56914 dataset (10 CTEPH whole blood samples and 10 control samples) was downloaded from the Gene Expression Omnibus database. Using limma package, the differentially expressed miRNAs (DE-miRNAs) were acquired. After miRNA-target pairs were obtained using miRWalk2.0 tool, a miRNA-target regulatory network was built by Cytoscape software. Using DAVID tool, significantly enriched pathways involving the target genes were identified. Moreover, the protein-protein interaction network and transcription factor-target regulatory network were built by the Cytoscape software. Additionally, quantitative real-time PCR (qRT-PCR) experiments and luciferase assay were conducted to validate miRNA/gene expression and miRNA-target regulatory relationship, respectively.
There were 25 DE-miRNAs (8 up-regulated and 17 down-regulated) between CTEPH and control groups. The target genes of has-let-7b-3p, has-miR-17-5p, has-miR-3202, has-miR-106b-5p, and has-miR-665 were enriched in multiple pathways such as "Insulin secretion". qRT-PCR analysis confirmed upregulation of hsa-miR-3202, hsa-miR-665, and matrix metalloproteinase 2 () as well as downregulation of hsa-let-7b-3p, hsa-miR-17-5p, and hsa-miR-106b-5p. Luciferase assay indicated that was negatively mediated by hsa-miR-106b-5p.
These miRNAs and genes were associated with the pathogenesis of CTEPH. Besides, hsa-miR-106b-5p was involved in the development of CTEPH via targeting .
慢性血栓栓塞性肺动脉高压（CTEPH）的特征是肺动脉压力升高。本研究旨在探索影响CTEPH发病机制的关键微小RNA（miRNA）和基因。
从基因表达综合数据库下载GSE56914数据集（10份CTEPH全血样本和10份对照样本）。使用limma软件包获取差异表达的miRNA（DE-miRNA）。使用miRWalk2.0工具获得miRNA-靶标对后，通过Cytoscape软件构建miRNA-靶标调控网络。使用DAVID工具鉴定涉及靶基因的显著富集通路。此外，通过Cytoscape软件构建蛋白质-蛋白质相互作用网络和转录因子-靶标调控网络。另外，分别进行定量实时PCR（qRT-PCR）实验和荧光素酶测定以验证miRNA/基因表达和miRNA-靶标调控关系。
CTEPH组和对照组之间有25个DE-miRNA（8个上调和17个下调）。has-let-7b-3p、has-miR-17-5p、has-miR-3202、has-miR-106b-5p和has-miR-665的靶基因在多个通路如“胰岛素分泌”中富集。qRT-PCR分析证实hsa-miR-3202、hsa-miR-665和基质金属蛋白酶2（）上调，以及hsa-let-7b-3p、hsa-miR-17-5p和hsa-miR-106b-5p下调。荧光素酶测定表明 由hsa-miR-106b-5p负向介导。
这些miRNA和基因与CTEPH的发病机制相关。此外，hsa-miR-106b-5p通过靶向 参与CTEPH的发展。
