Mir-141靶向结合ZEB1调控肺成纤维细胞的增殖和凋亡  

Mir-141 regulates proliferation and apoptosis of lung fibroblasts by targeting ZEB1

作　　者：杨益秀 孙娟[1] 周小曼[1] 李全妮 吴慈冰 赵洁 柳建芳 冯琼 丁毅鹏[1] YANG Yi-xiu;SUN Juan;ZHOU Xiao-man;LI Quan-ni;WU Ci-bing;ZHAO Jie;LIU Jian-fang;FENG Qiong;DING Yi-peng(Department of General Medicine,Hainan Provincial People’s Hospital,Haikou 570311,China;Hainan Hospital Affiliated to Hainan Medical University,Haikou 570311,China)

机构地区：[1]海南省人民医院全科医学科,海南海口570311 [2]海南医学院附属海南医院,海南海口570311

出　　处：《海南医学院学报》2019年第22期1693-1697,1702,共6页Journal of Hainan Medical University

基　　金：国家自然科学基金资助项目(81160008、81660013)~~

摘　　要：目的:研究mir-141在慢性阻塞性肺疾病(chronic obstructive pulmonary diseases,COPD)单个核细胞表达的意义,证明其对纤维细胞增殖凋亡的影响及靶向锌指蛋白E盒结合同源异形盒1蛋白(zinc finger E-box binding homeobox 1,ZEB1)的可能。方法:收集在我院40例COPD急性期,80例COPD稳定期病人和110例正常人对照的单个核细胞,RT-PCR检测mir-141并统计分析。构建mir-141 mimic,转染肺成纤维细胞,RT-PCR检测mir-141表达情况,CCK8检测细胞增殖情况,流式细胞术检测细胞凋亡,Targetscan预测ZEB1与mir-141的靶向关系,并使用RT-PCR与western blot进行验证。结果:COPD急性期与稳定期单个核细胞mir-141的表达显著高于正常对照组,而在COPD急性期显著高于COPD稳定期。使用mir-141 mimic转染后,mir-141显著上调,细胞活性明显促进,细胞凋亡率下降。Targetscan数据库预测ZEB1与mir-141有结合位点,双荧光素酶实验证实ZEB1与mir-141存在结合位点结合。使用mir-141 mimic转染后,ZEB1 mRNA与蛋白表达均下调。结论:COPD中高表达的mir-141可能通过靶向结合ZEB1促进肺成纤维细胞增殖,抑制凋亡。Objective:To study the significance of mir-141 expression in COPD(chronic obstructive pulmonary diseases)mononuclear cells,and to demonstrate its effect on proliferation and apoptosis of fibroblasts Possible to ZEB1(zinc finger E-box binding homeobox 1).Methods:A total of 40 cases acute phase of COPD patients,80 cases of stable period COPD patients and 110 normal controls in our hospital were collected.RT-PCR was used to detect mir-141 and statistical analysis.The mir-141 mimic was constructed and transfected into lung fibroblasts.The expression of mir-141 was analyzed by RT-PCR.Cell proliferation was analyzed by CCK8 and cell apoptosis was detected by flow cytometry.Targetscan was used to predict the binding site of ZEB1 and mir-141.The target relationship between ZEB1 and mir-141 was identified by RT-PCR and western blot.Results:The expression of mir-141 in mononuclear cells of acute phase of COPD and stable period of COPD was significantly higher than that of normal controls.Mir-141 levels in acute phase of COPD were significantly higher than that of stable period of COPD.Mir-141 mimic transfection significantly up-regulated the expression of mir-141,promoted cell activity and decreased cell apoptosis rate compared with the empty control group.ZEB1 and mir-141 had binding sites predicted by Targetscan database and verified by dual luciferase assays.After mir-141 mimic transfection hat,ZEB1 mRNA and protein expression were down-regulated.Conclusion:High expression of mir-141 in COPD may promote the proliferation of lung fibroblasts and inhibit apoptosis by targeting ZEB1.

关 键 词：COPD mir-141 ZEB1 增殖 凋亡 

分 类 号：R563[医药卫生—呼吸系统]