STAT3调控lncRNA HOTAIR介导炎症诱导EMT在吸烟所致COPD和肺癌中的作用及干预研究

Cigarette smoking is one of unhealthy lifestyles that threatens seriously human's health. The chromic obstructive pulmonary disease (COPD) and lung cancer are the most common and serious diseases induced by smoking, respectively, however, their molecular mechanisms remain unclear. Long non-coding RNA (lncRNA) has many important biological functions, but there are very few investigative publications about the roles and mechanisms of lncRNA in the damages induced by environmental pollution. Based on our preliminary results that CSE-induced inflammation, epithelial-mesenchymal transition (EMT), and the malignant transformation of human bronchial epithelial (HBE) cells, we are going to investigate the roles and interventions of inflammation to EMT mediated by STAT3-regulated lncRNA HOTAIR in cigarette smoking-induced COPD and lung cancer on the levels of cell, animal, and population, respectively, by using the following three models. (i) Cell model, treatmemt of HBE cells by CSE in the presence or absence of IL-6-neutralizing antibody, STAT3 inhibitor or siRNA, lncRNA HOTAIR siRNA or plasmid, Tan IIA, and Genistein, respectively; (ii) The Bal/c mice and Wistar rat models of COPD and tumorigenesis induced by cigarette smoke in the present or absent of Tan IIA or Genistein; (iii) Cohort study on population with different smoking levels. Our results will be helpful to understand the molecular mechanisms underlying smoking-induced COPD and lung cancer, which will provide new clues to find the early biomarkers and new measures of preventing and controling the smoking-induced lung damage.

吸烟是一种严重危害人类健康的不良生活方式；慢性阻塞性肺病(COPD)和肺癌是吸烟最常见和最严重的疾病，但其发病机制不清楚。长链非编码RNA(lncRNA)具有重要生物学功能，而其在环境污染物所致损伤中作用及机制研究报道甚少。本项目在已建立香烟烟雾抽提物(CSE)所致肺HBE细胞炎症、上皮间质转化(EMT)和恶性转化模型基础上，分别应用IL-6中和抗体、STAT3抑制剂或siRNA、正义或反义lncRNA HOTAIR及丹参酮IIA和三羟基异黄酮等处理CSE所致HBE细胞急性损伤和恶性转化模型，香烟烟雾所致COPD和肺癌动物模型及不同程度吸烟人群队列，从正向和反向在细胞、动物和人群探讨STAT3调控lncRNA HOTAIR介导炎症诱导EMT在吸烟所致COPD和肺癌中作用及干预效果，以揭示吸烟所致COPD和肺癌的部分分子机制、为寻找吸烟所致肺损伤的早期生物标志及发现新的防控措施提供科学依据。

吸烟是一种严重危害人类健康的不良生活方式，其成为全世界广泛关注的公共卫生问题之一，而慢性阻塞性肺病（COPD）和肺癌是吸烟最常见和最严重的疾病，由于其发病机制不清楚，目前对吸烟所致肺损害尚无有效的预防和治疗药物。本项目利用香烟抽提物（CSE）急性和慢性处理人支气管上皮（HBE）细胞所致上皮间质转化（EMT）和肿瘤干细胞（CSCs）特性获得及恶性转化；肺成纤维（MRC-5）细胞与CSE处理HBE细胞共培养及被外泌体处理等细胞实验；不同程度香烟烟雾慢性处理所致小鼠COPD动物实验；不同程度吸烟和COPD人群队列研究。运用多种分子生物学和毒理学方法在细胞、动物和人群三个水平系统地探讨了吸烟所致肺组织发生EMT、CSCs特性获得、肺组织炎症、气道重塑和COPD及肺癌的分子机制，以及穿心莲内脂对吸烟所致以上损害的拮抗效果和机制。.研究结果发现：长链非编码RNA（lncRNA）HOTAIR介导EMT和CSCs在CSE所致HBE细胞恶性转化中具有重要作用；lncRNA HOTAIR通过EZH2/H3K27me3表观下调p21参与CSE所致肺HBE细胞周期紊乱过程；miR-217转录后调控lncRNA MALAT1通过EZH2/H3K27me3参与了CSE所致HBE细胞发生EMT和恶性转化过程；lncRNA CCAT1表观调控miR-218和c-Myc与let-7c相互调控在CSE所致HBE细胞恶性转化中发挥重要作用；STAT3调控外泌体miR-21促进血管生成及其影响CSE所致恶性转化细胞的恶性程度和侵袭转移能力；HIF-1α调控miR-21通过靶向PTEN而激活Akt/NF-κB通路参与CSE所致HBE细胞恶性转化；miR-218通过靶向TNFR1调控NF-B在CSE所致气道炎症及黏液高分泌中具有重要作用；支气管上皮细胞分泌的外泌体miR-21促进成纤维细胞分化在吸烟所致气道重塑和COPD中具有重要作用；穿心莲内脂通过IL-6/STAT3下调lncRNA HOTAIR阻滞炎症诱导EMT发生而拮抗吸烟所致气道重塑和COPD。.本项目获得的研究结果揭示了吸烟所致COPD和肺癌发生发展的部分分子机制及穿心莲内脂的拮抗效果和机制，从而为寻找吸烟所致肺损害的早期生物标志和发现新的防治措施提供新线索，其对于吸烟所致机体损害防控具有重要的科学意义和一定的应用价值。