Basic Medicine College, Henan University of Chinese Medicine, Zhengzhou, China.
Henan Key Laboratory of Chinese Medicine for Respiratory Disease, Henan University of Chinese Medicine, Zhengzhou, China.
J Cell Biochem. 2019 Apr;120(4):6223-6236. doi: 10.1002/jcb.27910. Epub 2018 Oct 18.
Chronic obstructive pulmonary disease (COPD) is a leading cause of death due to tis high morbidity and mortality. microRNAs have emerged as new biomarkers for the prognosis and diagnosis of patients with COPD. In this study, we aimed to investigate the expression of microRNA-206 (miR-206) in lung tissues from COPD patients and to explore the regulatory role of miR-206 in the human pulmonary microvascular endothelial cells (HPMECs). Our results showed that cigarette smoke extract (CSE) promoted cell apoptosis, increased caspase-3 activity, and upregulated the expression of miR-206 in HPMECs, which was significantly reversed by the miR-206 knockdown. Transfection with miR-206 mimics led to cell apoptosis and was closely related to changes in the protein expression levels of caspase-3, caspase-9, and Bcl-2 in HPMECs. Further bioinformatics prediction analysis revealed that the 3'-untranslated region (3'UTR) of Notch3 and vascular endothelial growth factor-A (VEGFA) harbored miR-206-binding sites, and overexpression of miR-206 repressed the luciferase activity of the vectors containing Notch3 and VEGFA 3'UTR. Overexpression of either Notch3 or VEGFA attenuated miR-206-induced cell apoptosis in HPMECs. More importantly, miR-206 expression was upregulated in the lung tissues from COPD patients and was positively corrected with forced expiratory volume 1% predicted in COPD patients, while Notch3 and VEGFA mRNA levels were downregulated and were negatively correlated with the expression level of miR-206 in the lung tissues from COPD patients. In conclusion, our results showed that miR-206 was upregulated in COPD patients and CSE-treated HPMECs, promoted cell apoptosis via directly targeting Notch3 and VEGFA in HPMECs.
慢性阻塞性肺疾病（COPD）是导致高发病率和死亡率的主要死亡原因。 microRNAs 已成为 COPD 患者预后和诊断的新型生物标志物。在这项研究中，我们旨在研究 COPD 患者肺组织中 microRNA-206（miR-206）的表达，并探讨 miR-206 对人肺微血管内皮细胞（HPMEC）的调节作用。我们的结果表明，香烟烟雾提取物（CSE）促进细胞凋亡，增加 caspase-3 活性，并上调 HPMEC 中 miR-206 的表达，而 miR-206 的敲低则显著逆转了这一作用。转染 miR-206 模拟物导致细胞凋亡，与 HPMEC 中 caspase-3、caspase-9 和 Bcl-2 蛋白表达水平的变化密切相关。进一步的生物信息学预测分析表明，Notch3 和血管内皮生长因子 A（VEGFA）的 3'非翻译区（3'UTR）含有 miR-206 结合位点，并且 miR-206 的过表达抑制了含有 Notch3 和 VEGFA 3'UTR 的载体的荧光素酶活性。Notch3 或 VEGFA 的过表达减弱了 miR-206 诱导的 HPMEC 细胞凋亡。更重要的是，COPD 患者肺组织中 miR-206 的表达上调，并与 COPD 患者的用力呼气量 1%预测值呈正相关，而 Notch3 和 VEGFA mRNA 水平下调，并与 COPD 患者肺组织中 miR-206 的表达水平呈负相关。总之，我们的结果表明，miR-206 在 COPD 患者和 CSE 处理的 HPMEC 中上调，通过直接靶向 HPMEC 中的 Notch3 和 VEGFA 促进细胞凋亡。
