Department of Respiratory Medicine, The Hospital of Lianyungang Affiliated Xuzhou Medical University, Lianyungang, Jiangsu, China.
Department of Emergency, Nanjing Drum Tower Hospital Affiliated to Nanjing University Medical School, Nanjing, Jiangsu, China.
Biotechnol Appl Biochem. 2022 Apr;69(2):735-748. doi: 10.1002/bab.2148. Epub 2021 Apr 3.
Chronic obstructive pulmonary disease (COPD) is a chronic airway disorder mainly resulted from cigarette smoke exposure. The dysregulated circular RNAs (circRNAs) are relevant to the pathogenesis of COPD. This study aims to explore the function and mechanism of circRNA hsa_circ_0006892 (circ_0006892) in cigarette smoke extract (CSE)-induced bronchial epithelial injury. The lung tissues were collected from 17 nonsmokers and 23 smokers with COPD. The bronchial epithelial cells (BEAS-2B and 16HBE) were stimulated via CSE. Circ_0006892, microRNA-24 (miR-24), and PH domain and leucine-rich repeat protein phosphatase 2 (PHLPP2) abundances were examined via a quantitative reverse transcription polymerase chain reaction or Western blot. Cell viability, apoptosis, and inflammatory response were assessed via cell counting kit-8 (CCK-8), flow cytometry, and enzyme-linked immunosorbent assay (ELISA). The target relationship of miR-24 and circ_0006892 or PHLPP2 was tested via dual-luciferase reporter analysis, RNA immunoprecipitation, and RNA pull-down. Circ_0006892 expression was reduced in lung tissues of smokers with COPD and CSE-stimulated bronchial epithelial cells. Circ_0006892 overexpression alleviated CSE-induced viability reduction and promotion of apoptosis and inflammatory response. MiR-24 was bound via circ_0006892, and miR-24 overexpression reversed the effect of circ_0006892 on CSE-induced injury. PHLPP2 was targeted via miR-24, and miR-24 knockdown mitigated CSE-induced viability reduction and promotion of apoptosis and inflammatory response via regulating PHLPP2. Circ_0006892 could promote PHLPP2 expression via regulating miR-24. Circ_0006892 attenuated CSE-induced bronchial epithelial cell apoptosis and inflammatory response via regulating miR-24/PHLPP2 axis.
慢性阻塞性肺疾病（COPD）是一种主要由吸烟引起的慢性气道疾病。失调的环状 RNA（circRNA）与 COPD 的发病机制有关。本研究旨在探讨环状 RNA hsa_circ_0006892（circ_0006892）在香烟烟雾提取物（CSE）诱导的支气管上皮损伤中的作用和机制。收集了 17 名非吸烟者和 23 名 COPD 吸烟者的肺组织。用 CSE 刺激支气管上皮细胞（BEAS-2B 和 16HBE）。通过定量逆转录聚合酶链反应或 Western blot 检测 circ_0006892、microRNA-24（miR-24）和 PH 结构域和亮氨酸丰富重复蛋白磷酸酶 2（PHLPP2）的丰度。通过细胞计数试剂盒-8（CCK-8）、流式细胞术和酶联免疫吸附试验（ELISA）评估细胞活力、凋亡和炎症反应。通过双荧光素酶报告分析、RNA 免疫沉淀和 RNA 下拉测试验证 miR-24 与 circ_0006892 或 PHLPP2 的靶关系。COPD 吸烟者肺组织和 CSE 刺激的支气管上皮细胞中 circ_0006892 的表达减少。circ_0006892 过表达减轻了 CSE 诱导的活力降低以及促进凋亡和炎症反应。circ_0006892 结合 miR-24，而过表达 miR-24 逆转了 circ_0006892 对 CSE 诱导损伤的作用。PHLPP2 是 miR-24 的靶标，而 miR-24 敲低通过调节 PHLPP2 减轻 CSE 诱导的活力降低以及促进凋亡和炎症反应。circ_0006892 通过调节 miR-24 促进 PHLPP2 表达。circ_0006892 通过调节 miR-24/PHLPP2 轴减轻 CSE 诱导的支气管上皮细胞凋亡和炎症反应。
