抑制miR-155的表达对COPD患者气道平滑肌细胞增殖和迁移的影响  

Effect of inhibiting miR-155 expression on proliferation and migration of airway smooth muscle cell in patients with COPD

作　　者：王伟[1] 赵四林[1] 范伏元[1] 金朝晖[1] 李妲[1] 符艳 孙爽 肖雪飞 张慧卿 胡立 WANG Wei;ZHAO Si-lin;FAN Fu-yuan;JIN Zhao-hui;LI Da;FU Yan;SUN Shuang;XIAO Xue-fei;ZHANG Hui-qing;HU Li(Department of Respiratory Medicine,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410007,China;Department of Respiratory Medicine,Hunan Provincial People's Hospital,Changsha 410007,China)

机构地区：[1]湖南中医药大学第一附属医院呼吸内科,湖南长沙410007 [2]湖南省人民医院呼吸内科,湖南长沙410007

出　　处：《海南医学院学报》2021年第10期763-767,共5页Journal of Hainan Medical University

基　　金：湖南省中医药科研计划项目(201838)。

摘　　要：目的:探讨抑制miR-155表达对慢性阻塞性肺病(COPD)患者气道平滑肌细胞(ASMCs)增殖和迁移的影响。方法:分离培养来自8例COPD患者(观察组)和3例无COPD病史肺部良性肿瘤患者(对照组)的ASMCs。将miR-155抑制表达质粒(anti-miR-155)和阴性对照质粒(anti-miR-NC)转染至ASMCs,同时设置空白组。采用实时荧光定量PCR(RT-qPCR)检测miR-155表达水平;流式细胞仪检测细胞周期;克隆形成实验检测细胞克隆形成数;Transwell检测细胞迁移和侵袭;酶联免疫吸附试验(ELISA)法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平。结果:观察组ASMCs中miR-155表达水平明显高于对照组(P<0.05)。抑制miR-155表达组miR-155表达水平明显低于阴性对照组和空白组(P<0.05)。抑制miR-155表达组,G0-G1期细胞所占比例升高,S期细胞比例而降低,细胞克隆数及迁移、侵袭细胞数降低,TNF-α和IL-6水平升高(P<0.05)。结论:抑制miR-155的表达可抑制COPD患者气道平滑肌细胞增殖、迁移,抑制促炎因子的释放。Objective:To investigate the effect of inhibiting miR-155 expression on the proliferation and migration of airway smooth muscle cells(ASMCs)in patients with chronic obstructive pulmonary disease(COPD).Methods:ASMCs were isolated and cultured from 8 patients with COPD(observation group)and 3 patients with benign lung cancer without COPD(control group).The ASMCs were transfected with miR-155 suppression expression plasmid(anti-miR-155)and the negative control plasmid(anti-miR-NC),and blank control group was set.Real-time quantitative PCR(RT-qPCR)was used to detect the expression of miR-155;flow cytometry was used to detect the cell cycle;clone formation experiment was used to detect the number of cell clones;Transwell was used to detect cell migration and invasion;Enzyme-linked immunosorbent assay(ELISA)method was used to detect tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)level.Results:The expression level of miR-155 in ASMCs of observation group was significantly higher than that in the control group(P<0.05).The miR-155 expression level in inhibited miR-155 expression group was significantly lower,compared with the negative control group and the blank group(P<0.05).In the inhibited miR-155 expression group,the proportion of G0-G1 phase cells increased,the proportion of S phase cells decreased,the number of cell clones,migration,and the number of invasive cells decreased,and the levels of TNF-αand IL-6 increased(P<0.05).Conclusions:Inhibiting the expression of miR-155 can inhibit the proliferation and migration of airway smooth muscle cells in COPD patients,and inhibit the release of proinflammatory factors.

关 键 词：慢性阻塞性肺病 MIR-155 气道平滑肌细胞 增殖 迁移 

分 类 号：R563.9[医药卫生—呼吸系统]