METTL14上调miR-141的m6A修饰抑制ZEB1促进肺成纤维细胞增殖与炎症  

METTL14 upregulates m6A modification of pri-miR-141 inhibiting ZEB1 to promote proliferation and inflammation of lung fibroblasts

作　　者：杨益秀 陈洁 李全妮 林梅 丁毅鹏[1,2] YANG Yi‑xiu;CHEN Jie;LI Quan-ni;LIN Mei;DING Yi‑peng(Hainan Hospital Affiliated to Hainan Medical University,Haikou 570311,China;Department of General Practice,Hainan General Hospital,Haikou 570311,China)

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

出　　处：《海南医学院学报》2023年第2期123-128,共6页Journal of Hainan Medical University

基　　金：海南省卫生健康行业科研项目(21A200259)。

摘　　要：目的:探究METTL14是否通过调控pri-miR-141的m6A修饰参与调控成纤维细胞的增殖和炎症因子分泌。方法:使用METTL14过表达慢病毒转染MRC-5细胞,采用qPCR和western blot检测METTL14和ZEB1的表达量;采用CCK-8和流式细胞术检测METTL14对MRC-5增殖和凋亡的影响;IL-2,IL6和TNF-α的ELISA试剂盒检测METTL14对MRC-5分泌炎症因子的影响;采用meRIP检测pri-miR-141上的m6A修饰位点,RIP检测pri-miR-141与METTL14的结合情况。结果:在MRC-5细胞中成功过表达METTL14基因;METTL14高表达促进MRC-5细胞增殖,抑制其凋亡,并促进MRC-5细胞分泌炎症因子;pri-miR-141上存在m6A修饰位点,且pri-miR-141能够和METTL14直接结合;METTL14高表达增加miR-141的表达量,并抑制ZEB1 mRNA与蛋白表达。结论:METTL14通过增加pri-miR-141的m6A修饰位点促进miR-141的表达,抑制ZEB1从而促进成纤维细胞的增殖和炎症因子的分泌。Objective:To explore whether METTL14 is involved in regulating the fibroblast proliferation and inflammatory cytokine secretion by regulating the m6A modification of pri-miR-141.Methods:MRC-5 cells were transfected via METTL14 overexpression lentivirus to increase METTL14 expression.Levels of METTL14 and ZEB1 were measured by qPCR and western blot.The effect of METTL14 on MRC-5 proliferation and apoptosis was determined by CCK-8 and flow cytometry,respectively.The ELISA kits of IL-2,IL6 and TNF-αwere used to detect the effect of METTL14 on MRC-5 inflammatory secretion.m6A modification site on pri-miR-141 was detected by meRIP.The binding site between pri-miR-141 and METTL14 was determined by RIP.Results:We successfully upregulated METTL14 expression in MRC-5 cells.Elevated METTL14 promoted MRC-5 cell proliferation,suppressed its apoptosis and promoted inflammatory factors secretion in MRC-5 cells.pri-miR-141 had m6A modification sites.pri-miR-141 can directly bind to METTL14.METTL14 upregulation increased miR-141 while suppressed ZEB1 expression.Conclusion:METTL14 can promote the expression of miR-141 by increasing the m6A modification site of pri-miR-141,and inhibit ZEB1,thereby promoting the proliferation of fibroblasts and the secretion of inflammatory factors.

关 键 词：成纤维细胞 增殖 炎症因子 METTL14 m6A 

分 类 号：R563.9[医药卫生—呼吸系统]