LPS处理肺泡上皮A549细胞诱发微RNA-21表达下降及其靶向调控PDCD4减少细胞凋亡的实验研究  

LPS-treated alveolar epithelial cell line A549 induces down-regulation of miR-21 expression and its targeting regulation of PDCD4 to reduce apoptosis

作　　者：张德振[1] 吴涛[2] 李志杰[1] Zhang Dezhen;Wu Tao;Li Zhijie(Department of Respiratory and Critical Care Medicine, Shangqiu First Municipal People’s Hospital, Shangqiu, Henan 476100, China;Department of Interventional Medicine, Henan University of Traditional Chinese Medicine, Zhengzhou, Henan 450000, China)

机构地区：[1]商丘市第一人民医院呼吸与危重症医学科,河南476100 [2]河南中医药大学介入科,郑州45000

出　　处：《中华生物医学工程杂志》2019年第3期318-322,共5页Chinese Journal of Biomedical Engineering

摘　　要：目的研究微RNA(miR)-21在脂多糖(LPS)诱导的肺泡上皮A549细胞凋亡中的作用。方法用LPS处理A549细胞,qRT-PCR检测miR-21表达变化。在A549细胞中转染miR-21 mimics,MTT测定细胞增殖,流式细胞术检测细胞凋亡,免疫印迹检测C-capase-3蛋白表达水平,使用靶基因预测软件预测程序性细胞死亡4(PDCD4)可能是miR-21的靶基因,在A549细胞中共转染miR-21 mimics、pcDNA3.1-PDCD4,用上述方法测定细胞增殖、凋亡和C-capase-3蛋白表达变化。结果LPS处理后的A549细胞中miR-21表达水平降低。LPS处理后的A549细胞增殖活性降低,细胞凋亡率和C-capase-3蛋白水平升高。miR-21 mimics可以提高LPS条件下A549细胞增殖活性,降低细胞凋亡率和C-capase-3蛋白水平。miR-21靶向负调控PDCD4表达。上调PDCD4抑制miR-21 mimics抗LPS诱导的A549细胞凋亡和增殖活性降低作用。结论LPS诱导A549细胞中miR-21表达下调,miR-21靶向调控PDCD4抑制LPS诱导的A549细胞凋亡发生。Objective To investigate the effect of microRNA (miR)-21 in lipopolysaccharide (LPS)-induced apoptosis of alveolar epithelial cell line A549. Methods A549 cells were treated with LPS, and the change in expression of miR-21 was determined by qRT-PCR. miR-21 mimics were transfected into A549 cells. The cell proliferation was measured by MTT, and the cell apoptosis was examined by flow cytometry. Western blotting was used to determine the expression level of C-capase-3 protein.The target gene prediction software predicted that programmed cell death protein 4 (PDCD4) might be the target gene of miR-21. Then, miR-21 mimics and pcDNA3.1-PDCD4 were co-transfected into A549 cells. The cell proliferation, apoptosis and change in C-capase-3 protein expression were measured by the above methods. Results The expression level of miR-21 decreased in LPS-treated A549 cells. The proliferation activity of A549 cells decreasedafter LPS treatment, and the apoptosis rate and C-capase-3 protein level increased. miR-21 mimics increased the proliferation activity of A549 cells, and decreased the apoptosis rate and C-capase-3 protein level under LPS conditions. miR-21 targeted negative regulation of PDCD4 expression. Up-regulation of PDCD4 inhibited the anti-LPS-induced apoptosis and proliferation of A549 cells by miR-21 mimics. Conclusion LPS induces down-regulation of miR-21 expression in A549 cells, and miR-21 targeting regulation of PDCD4 inhibits LPS-induced apoptosis in A549 cells.

关 键 词：肺泡上皮细胞 微RNA-21 凋亡 PDCD4 脂多糖 

分 类 号：R563.8[医药卫生—呼吸系统]