Journal Article

Expression of long non‐coding RNA LUCAT1 in patients with chronic obstructive pulmonary disease and its potential functions in regulating cigarette smoke extract‐induced 16HBE cell proliferation and apoptosis

Lu, Junjie, Cheng, Jing, Yang, Tao, Lin, Chunyan, Qian, Xiaoyu, Zhao, Shan

2021

Overview

Background Chronic obstructive pulmonary disease (COPD), characterized by persistent airflow limitation, was a disease mediated by a combination of inflammatory factors, immune cells, and immune mediators. COPD was an inflammatory and autoimmune disease involving T‐lymphocytes triggered by cigarette smoke and other factors that progressively affected the bronchi, lung parenchyma, and pulmonary blood vessels. LncRNAs were reported to be implicated in COPD pathogenesis and development. Methods Non‐smokers, smokers (non‐COPD), and COPD patients were randomly selected in an established COPD surveillance cohort. Demographic and clinical information of all subjects were collected. Pulmonary function was measured by post‐bronchodilator testing. qRT‐PCR and ELISA assays were performed to detect the expression levels of lncRNA LUCAT1, miR‐181a‐5p, and inflammatory cytokines. An in vitro exposure model was constructed using cigarette smoke extract (CSE)‐induced human bronchial epithelial (16HBE) cells. The dual‐luciferase reporter and RNA pull‐down assays were used to detect the binding relationship between lncRNA LUCAT1 and miR‐181a‐5p; meanwhile, Spearman's correlation assay was used to verify the correlation between lncRNA LUCAT1 and miR‐181a‐5p. Afterward, the lncRNA LUCAT1 silencing plasmid was constructed and co‐transfected with a miR‐181a‐5p inhibitor to evaluate the effects on CSE‐induced 16HBE cell proliferation and apoptosis. Finally, a Western blot assay was utilized to determine the mechanism of lncRNA LUCAT1/miR‐181a‐5p/Wnt/β‐catenin axis in COPD. Results LncRNA LUCAT1 was upregulated in the serums of COPD patients. Correlation analysis further confirmed the strong correlation between LUCAT1 expression and inflammatory cytokines IL‐1β, IL‐6, and TNF‐α. Receiver operating characteristic (ROC) analysis verified the potential of LUCAT1 in COPD diagnosis. After treatment with CSE, LUCAT1 was significantly increased while its target miR‐181a‐5p was decreased in 16HBE cells. Cell proliferation and apoptosis assays showed that LUCAT1 silencing alleviated CSE’s effects on 16HBE cell proliferation and apoptosis. Mechanically, rescue assays demonstrated that miR‐181a‐5p inhibition could partially counteract the impact of LUCAT1 on COPD progression through the Wnt/β‐catenin pathway. Conclusions LncRNA LUCAT1 may be a valuable indicator for differentiating COPD. Moreover, LncRNA LUCAT1/miR‐181‐5p/Wnt/β‐catenin axis behaved as a critical role in COPD development, shedding new sights for clinical treatment. Chronic obstructive pulmonary disease (COPD) was a chronic, non‐homogeneous lung disease characterized by persistent excessive inflammation that led to tissue remodeling, alveolar damage, airflow limitation, and accelerated lung function decline. The clinical manifestations of COPD include chronic cough, coughing sputum, dyspnea, and acute exacerbation in the presence of infections and other triggering factors. Pulmonary function tests were the primary means of confirming the diagnosis of COPD (Respiratory Care, 57, 2012 and 165). Appropriate drug therapy can effectively relieve symptoms, reduce the number and frequency of acute exacerbations, improve exercise tolerance, and improve quality of life. Cigarette smoking was a significant risk factor for COPD. Inhaled cigarette smoke first encountered respiratory epithelial cells, causing the release of pro‐inflammatory mediators such as interleukin (IL)‐1, IL‐6, and tumor necrosis factor (TNF)‐α. Although inflammation‐based airway injury was recognized as a core feature of COPD, the underlying mechanism was unclear. From the results, we found that lncRNA LUCAT1 was upregulated in the serums of COPD patients. Correlation analysis further confirmed the strong correlation between LUCAT1 expression and inflammatory cytokines IL‐1β, IL‐6, and TNF‐α. Receiver operating characteristic (ROC) analysis verified the potential of LUCAT1 in COPD diagnosis. After treatment with CSE, LUCAT1 was significantly increased, while its target miR‐181a‐5p was decreased in 16HBE cells. Cell proliferation and apoptosis assays showed that LUCAT1 silencing alleviated CSE’s effects on 16HBE cell proliferation and apoptosis. Mechanically, rescue assays demonstrated that miR‐181a‐5p inhibition could partially counteract the impact of LUCAT1 on COPD progression through the Wnt/β‐catenin pathway. In sum, lncRNA LUCAT1 may be a valuable indicator for differentiating COPD. Moreover, lncRNA LUCAT1/miR‐181‐5p/Wnt/β‐catenin axis behaved as a critical role in COPD development, shedding new sights for clinical treatment.

• Publisher: John Wiley & Sons, Inc,John Wiley and Sons Inc
• Subject: 16HBE / Aged / Apoptosis / Apoptosis - genetics / Autoimmune diseases / Base Sequence / Biomarkers - metabolism / Biotechnology / Blood vessels / Bronchodilators / Bronchus / Catenin / Cell Line / Cell proliferation / Cell Proliferation - genetics / Chronic obstructive pulmonary disease / Cigarette smoke / cigarette smoke extract / Cigarettes / Correlation analysis / Disease Progression / Down-Regulation - genetics / Enzyme-linked immunosorbent assay / Enzymes / Female / Gene Silencing / Humans / lncRNA LUCAT1 / Lung diseases / Lymphocytes / Male / MicroRNAs - genetics / MicroRNAs - metabolism / miR‐181a‐5p / Non-coding RNA / Obstructive lung disease / Parenchyma / Patients / Proteins / Pulmonary Disease, Chronic Obstructive - diagnosis / Pulmonary Disease, Chronic Obstructive - genetics / Pulmonary Disease, Chronic Obstructive - pathology / Respiratory function / RNA, Long Noncoding - metabolism / Smoking / Smoking - genetics / Tumor necrosis factor / Tumor necrosis factor-TNF / Up-Regulation - genetics / Wnt protein / Wnt Signaling Pathway - genetics