Department of Critical Care Medicine, First People's Hospital of Jining City, Jining City, Shandong Province, China.
First People's Hospital of Jining City Affiliated to Jining Medical College, Jining City, Shandong Province, China.
J Mol Histol. 2021 Jun;52(3):437-447. doi: 10.1007/s10735-021-09972-2. Epub 2021 Apr 13.
Chronic obstructive pulmonary disease (COPD) is an inflammatory respiratory disease. Long non-coding RNAs (lncRNAs) have been implicated in the pathogenesis of COPD. In the present study, we set to investigate the role and mechanism of LOC729178 on cigarette smoke extract (CSE)-induced inflammatory damage in 16HBE cells. The expression levels of LOC729178, miR-144-3p, and PH domain leucine-rich repeat protein phosphatase 2 (PHLPP2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell viability and apoptosis were assessed by Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. Enzyme-linked immunosorbent assay (ELISA) assay was performed to evaluate the levels of interleukin-1β (IL-1β), IL-6, tumor necrosis factor-alpha (TNF-α), and IL-8. Targeted relationships among LOC729178, miR-144-3p, and PHLPP2 were verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Our data indicated that LOC729178 was underexpressed in COPD tissues and CSE-treated 16HBE cells. Exogenous expression of LOC729178 alleviated CSE-induced inflammatory injury in 16HBE cells. LOC729178 targeted miR-144-3p by directly binding to miR-144-3p. miR-144-3p was a downstream effector of LOC729178 function. PHLPP2 was identified as a direct and functional target of miR-144-3p. Furthermore, LOC729178 operated as a post-transcriptional regulator of PHLPP2 expression through miR-144-3p. Our current study suggested that LOC729178 overexpression alleviated CSE-induced inflammatory injury in 16HBE cells at least in part by up-regulating PHLPP2 via sponging miR-144-3p, providing a rationale for developing LOC729178 as a potential therapeutic agent against COPD.
慢性阻塞性肺疾病（COPD）是一种炎症性呼吸系统疾病。长链非编码 RNA（lncRNA）已被认为与 COPD 的发病机制有关。在本研究中，我们旨在研究 LOC729178 在香烟烟雾提取物（CSE）诱导的 16HBE 细胞炎症损伤中的作用和机制。通过定量实时聚合酶链反应（qRT-PCR）和 Western blot 检测 LOC729178、miR-144-3p 和 PH 结构域亮氨酸丰富重复蛋白磷酸酶 2（PHLPP2）的表达水平。通过细胞计数试剂盒-8（CCK-8）和流式细胞术分别评估细胞活力和细胞凋亡。酶联免疫吸附试验（ELISA）测定白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和白细胞介素-8（IL-8）的水平。通过双荧光素酶报告和 RNA 免疫沉淀（RIP）实验验证 LOC729178、miR-144-3p 和 PHLPP2 之间的靶向关系。我们的数据表明，LOC729178 在 COPD 组织和 CSE 处理的 16HBE 细胞中表达下调。外源性表达 LOC729178 减轻了 CSE 诱导的 16HBE 细胞炎症损伤。LOC729178 通过直接结合 miR-144-3p 靶向 miR-144-3p。miR-144-3p 是 LOC729178 功能的下游效应物。PHLPP2 被鉴定为 miR-144-3p 的直接和功能靶标。此外，LOC729178 通过 miR-144-3p 作为 PHLPP2 表达的转录后调节剂起作用。我们的研究表明，LOC729178 通过海绵吸附 miR-144-3p 来上调 PHLPP2，从而至少部分缓解 CSE 诱导的 16HBE 细胞炎症损伤，为将 LOC729178 作为 COPD 的潜在治疗药物提供了依据。
