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 (PIK3R2) 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.
方法:“快速”进展性COPD组包括6例,“缓慢”和正常进行性COPD组各5例,COPD组3例。外周血样本取自参与者,其次是总RNA提取和高通量miRNA测序。使用生物信息学分析鉴定进展性COPD组中差异表达的miRNA。然后,候选miRNA进行外部验证。此外,该miRNA的靶基因被鉴定出来,以及它对细胞活性的影响,细胞周期,凋亡,分析COPD的其他生物学表型。
结果:与正常组相比,在“快速”进行性COPD中鉴定出总共35、16和7种差异表达的miRNA,“缓慢”进行性COPD组,COPD组,分别。使用双荧光素酶报告基因测定和使用磷酸肌醇-3-激酶的转染测试进一步证实了结果。调节亚基2(PIK3R2)作为miR-4433a-5p的靶基因;结果显示miRNA与其靶基因之间存在负调节相关性。表型检测表明,磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路的激活可能通过促进炎性A549细胞的增殖和抑制细胞凋亡参与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可以通过调节表型影响COPD的进展,如细胞增殖和凋亡。