Department of Preventive Medicine, Shantou University Medical College, Shantou, Guangdong 515041, People's Republic of China.
Center for Research and Technology of Precision Medicine, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen, Guangdong 518055, People's Republic of China.
Int J Chron Obstruct Pulmon Dis. 2019 Oct 9;14:2319-2329. doi: 10.2147/COPD.S224763. eCollection 2019.
Chronic obstructive pulmonary disease (COPD) is recognized as a chronic lung disease with incomplete reversible airflow limitation, but its pathophysiology was still not clear. This study aimed at investigating regulatory roles of special miRNA-mRNA axis in COPD development.
Differentially expressed miRNAs and downstream mRNAs were screened from the Gene Expression Omnibus (GEO) dataset by using the LIMMA package in R software. Weighted Gene Co-expression Network Analysis (WGCNA) was used to construct a co-expression network for COPD. The correlation of dysregulated miRNA(s) and COPD was analyzed, and miRNAs with significant differences were validated in peripheral blood mononuclear cells (PBMCs) from COPD patients by real-time PCR. Regulatory roles of candidate miRNAs and targeted mRNAs were investigated in vitro study.
Thirteen modules of co-expressed miRNAs and mRNAs were constructed from a selected cohort with WGCNA. Turquoise module with 12 differentially expressed miRNAs and 120 mRNAs was significantly correlated with COPD. The expression of hsa-miR-664a-3p, an upregulated miRNA in the module, was increased both in lung tissue and PBMCs from COPD patients, whereas that targeted four and a half LIM domains 1 () gene was decreased and positively correlated with forced expiratory volume in 1 sec (FEV1)/forced vital capacity (FVC%) ( = 0.59, < 0.01). In vitro, luciferase activity assay revealed as a target of hsa-miR-664a-3p and it could be directly downregulated by overexpression of hsa-miR-664a-3p. Furthermore, cigarette smoke extract could increase hsa-miR-664a-3p level and decrease level in Beas-2B cells.
The present study validated significant upregulation of hsa-miR-664a-3p in COPD patients, and its target gene was downregulated and positively correlated with FEV1/FVC%; both hsa-miR-664a-3p and could be regulated by cigarette smoke extract. Results of bioinformatic analyses and expanded validation suggest that the axis from hsa-miR-664a-3p to might play a key role in cigarette smoke-induced COPD, and the exact mechanism should be confirmed in further studies.
慢性阻塞性肺疾病（COPD）被认为是一种具有不完全可逆气流受限的慢性肺部疾病，但它的病理生理学仍然不清楚。本研究旨在探讨 COPD 发展中特殊 miRNA-mRNA 轴的调节作用。
从基因表达综合数据库（GEO）中使用 R 软件中的 LIMMA 包筛选差异表达的 miRNA 和下游 mRNA。使用加权基因共表达网络分析（WGCNA）构建 COPD 的共表达网络。分析失调 miRNA 与 COPD 的相关性，并通过实时 PCR 验证 COPD 患者外周血单个核细胞（PBMC）中差异表达的 miRNA。在体外研究中，对候选 miRNA 和靶向 mRNAs 的调节作用进行了研究。
WGCNA 从选定队列构建了 13 个 miRNA 和 mRNAs 的共表达模块。 turquoise 模块有 12 个差异表达的 miRNA 和 120 个 mRNAs，与 COPD 显著相关。该模块中上调的 hsa-miR-664a-3p 在 COPD 患者的肺组织和 PBMCs 中均增加，而靶向四个半 LIM 结构域 1()的基因则减少，并与 1 秒用力呼气量（FEV1）/用力肺活量（FVC%）呈正相关（=0.59，<0.01）。体外实验，荧光素酶活性测定显示作为 hsa-miR-664a-3p 的靶点，并可被 hsa-miR-664a-3p 的过表达直接下调。此外，香烟烟雾提取物可增加 Beas-2B 细胞中 hsa-miR-664a-3p 的水平，并降低的水平。
本研究验证了 COPD 患者 hsa-miR-664a-3p 的显著上调，其靶基因下调并与 FEV1/FVC%呈正相关；hsa-miR-664a-3p 和均可受香烟烟雾提取物调节。生物信息学分析和扩展验证的结果表明，从 hsa-miR-664a-3p 到的轴可能在香烟烟雾诱导的 COPD 中发挥关键作用，确切的机制应在进一步的研究中证实。
