差异 miRNA 谱分析显示 miR-4433a-5p 通过 PIK3R2 介导的表型调节成为慢性阻塞性肺疾病进展的关键调节因子。
Differential miRNA Profiling Reveals miR-4433a-5p as a Key Regulator of Chronic Obstructive Pulmonary Disease Progression via PIK3R2- mediated Phenotypic Modulation.
机构信息
Department of Respiratory and Critical Care Medicine, Fourth Affiliated Hospital of Xinjiang Medical University, Urumqi, China.
Xinjiang Laboratory of Respiratory Disease Research, Traditional Chinese Medicine Hospital Affiliated to Xinjiang Medical University, Urumqi, China.
出版信息
Comb Chem High Throughput Screen. 2024;27(16):2323-2334. doi: 10.2174/0113862073243966231030093213.
OBJECTIVE
In this study, a high-throughput sequencing technology was used to screen the differentially expressed miRNA in the patients with "fast" and "slow" progression of chronic obstructive pulmonary disease (COPD). Moreover, the possible mechanism affecting the progression of COPD was preliminarily analyzed based on the target genes of candidate miRNAs.
METHODS
The "fast" progressive COPD group included 6 cases, "slow" and Normal progressive COPD groups included 5 cases each, and COPD group included 3 cases. The peripheral blood samples were taken from the participants, followed by total RNA extraction and high throughput miRNA sequencing. The differentially expressed miRNAs among the progressive COPD groups were identified using bioinformatics analysis. Then, the candidate miRNAs were externally verified. In addition, the target gene of this miRNA was identified, and its effects on cell activity, cell cycle, apoptosis, and other biological phenotypes of COPD were analyzed.
RESULTS
Compared to the Normal group, a total of 35, 16, and 7 differentially expressed miRNAs were identified in the "fast" progressive COPD, "slow" progressive COPD group, and COPD group, respectively. The results were further confirmed using dual-luciferase reporter assay and transfection tests with phosphoinositide- 3-kinase, regulatory subunit 2 () as a target gene of miR-4433a-5p; the result showed a negative regulatory correlation between the miRNA and its target gene. The phenotype detection showed that the activation of the phosphatidylinositol 3 kinase (PI3K)/protein kinase B (AKT) signaling pathway might participate in the progression of COPD by promoting the proliferation of inflammatory A549 cells and inhibiting cellular apoptosis.
CONCLUSIONS
MiR-4433a-5p can be used as a marker and potential therapeutic target for the progression of COPD. As a target gene of miR-4433a-5p, PIK3R2 can affect the progression of COPD by regulating phenotypes, such as cellular proliferation and apoptosis.
目的
本研究采用高通量测序技术筛选慢性阻塞性肺疾病（COPD）“快进展”和“慢进展”患者差异表达的 microRNA，基于候选 microRNA 的靶基因初步分析影响 COPD 进展的可能机制。
方法
“快进展”COPD 组纳入 6 例，“慢进展”和“正常进展”COPD 组各纳入 5 例，COPD 组纳入 3 例，采集研究对象外周血，提取总 RNA，进行高通量 miRNA 测序。采用生物信息学分析鉴定 COPD 进展组间差异表达的 microRNA，然后进行候选 microRNA 的外部验证。此外，鉴定该 microRNA 的靶基因，并分析其对 COPD 细胞活性、细胞周期、凋亡等生物学表型的影响。
结果
与正常组相比，“快进展”COPD、“慢进展”COPD 组和 COPD 组分别鉴定出 35、16 和 7 个差异表达的 microRNA。利用双荧光素酶报告基因检测和以磷酸肌醇 3-激酶，调节亚基 2（）为 miR-4433a-5p 靶基因的转染实验进一步验证，结果显示 microRNA 与靶基因之间存在负调控关系。表型检测显示，通过促进炎症 A549 细胞增殖和抑制细胞凋亡，激活磷脂酰肌醇 3 激酶（PI3K）/蛋白激酶 B（AKT）信号通路可能参与 COPD 的进展。
结论
miR-4433a-5p 可作为 COPD 进展的标志物和潜在治疗靶点。作为 miR-4433a-5p 的靶基因，PIK3R2 可通过调节细胞增殖和凋亡等表型影响 COPD 的进展。
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