LncRNA MALAT1通过募集EZH2介导系统性红斑狼疮相关抗炎基因表观遗传沉默  

LncRNA MALAT1 Mediates Epigenetic Silencing of Systemic Lupus Erythematosus-Associated Anti-Inflammatory Genes through Recruitment of EZH2

作　　者：高飞 谈园 张莲 罗立芳 罗雯 马乐 黎姣艳 黄敖 GAO Fei;TAN Yuan;ZHANG Lian(Changsha First Hospital,Hunan Changsha 410005,China)

机构地区：[1]湖南省长沙市第一医院,湖南长沙410005 [2]湖南省中医学院第一附属医院皮肤科,湖南长沙410005

出　　处：《河北医学》2024年第9期1434-1439,共6页Hebei Medicine

基　　金：湖南省卫生健康委科研计划项目,(编号:202104120051);长沙市科学技术局项目,(编号:kdz2401039)。

摘　　要：目的:探究系统性红斑狼疮(SLE)中长链非编码RNA(LncRNA)MALAT1和甲基化转移酶EZH2与SLE相关抗炎因子的相关性和分子调控机制。方法:提取健康对照(n=10)和SLE患者(n=10)外周血单个核细胞(PBMC),采用实时荧光定量聚合酶链式反应(RT-qPCR)检测LncRNA MALAT1、EZH2和抗炎因子PTEN、Ets-1、FOXO1的mRNA水平,并分别验证LncRNA MALAT1、EZH2与各抗压因子表达的相关性;使用小干扰RNA(si-RNA)转染THP-1细胞分别敲低LncRNA MALAT1、EZH2后RT-qPCR检测各抗炎因子表达,RNA免疫沉淀(RIP)验证LncRNA MALAT1和EZH2的相互作用;同时敲低EZH2和过表达LncRNA MALAT1后使用蛋白质免疫印迹(western blot)检测组蛋白3(H3)第27位赖氨酸(K27)三甲基化(me3)水平。结果:与健康对照相比,SLE患者外周血PBMC LncRNA MALAT1和EZH2显著高表达(P<0.05),抗炎因子PTEN、Ets-1、FOXO1表达显著低表达(P<0.05),且LncRNA MALAT1和EZH2和各抗炎表达显著负相关;分别敲低LncRNA MALAT1和EZH2后各抗炎因子表达均显著升高(P<0.05);RIP结果显示LncRNA MALAT1和EZH2存在相互结合;过表达LncRNA MALAT1后各抗炎因子表达显著降低,但是敲低EZH2同时过表达LncRNA MALAT1检测到各抗炎因子表达升高且H3K27me3显著降低(P<0.05)。结论:LncRNA MALAT1和EZH2表达与SLE相关抗炎因子表达显著负相关,其分子机制涉及LncRNA MALAT1通过招募EZH2调节H3K27me3表达相关。Objective:To explore the correlation of long non-coding RNA(LncRNA)MALAT1 and methyltransferase EZH2 with SLE-associated anti-inflammatory factors and the molecular regulatory mechanism.Methods:Peripheral blood mononuclear cells(PBMC)were extracted from healthy controls(n=10)and SLE patients(n=10),and mRNA levels of LncRNA MALAT1,EZH2,and anti-inflammatory factors PTEN,Ets-1,and FOXO1 were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).The correlation between LncRNA MALAT1,EZH2,and the expression of stress factors was verified respectively.After transfecting THP-1 cells with small interfering RNA(si-RNA),the expression of various anti-inflammatory factors was detected by RT-qPCR after LncRNA MALAT1 and EZH2 was knocked down,and the interaction between LncRNAMALAT1 and EZH2 was verified by RNA immunoprecipitation(RIP).After simultaneous knockdown of EZH2 and overexpression of LncRNAMALAT1,the 27th lysine(K27)trimethylation(me3)level of histone 3(H3)was detected by Western blot(western blot).Results:Compared with healthy controls,PBMC LncRNAMALAT1 and EZH2 in peripheral blood of SLE patients were significantly overexpressed(P<0.05),anti-inflammatory factors PTEN,Ets-1,and FOXO1 were significantly decreased(P<0.05),and LncRNA MALAT1 and EZH2 were significantly negatively correlated with the anti-inflammatory expressions.The expressions of various anti-inflammatory factors were significantly increased after LncRNA MALAT1 and EZH2 were knocked down respectively(P<0.05).The RIP result shows that LncRNA MALAT1 and EZH2 combine.After overexpression of LncRNA MALAT1,the expression of anti-inflammatory factors was significantly decreased.Still,the expression of anti-inflammatory factors was increased and H3K27me3 was decreased considerably when EZH2 was knocked down and LncRNA MALAT1 was overexpressed at the same time(P<0.05).Conclusion:The expression of LncRNA MALAT1 and EZH2 is significantly negatively correlated with the expression of SLE-associated anti-inflammatory factors,and the m

关 键 词：甲基化转移酶 表观遗传学沉默 系统性红斑狼疮 抗炎因子 

分 类 号：R593.241[医药卫生—内科学]