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OACSTPCD
LncRNA MALAT1 Mediates Epigenetic Silencing of Systemic Lupus Erythematosus-Associated Anti-Inflammatory Genes through Recruitment of EZH2
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 MAL-AT1、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 MAL-AT1 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 Ln-cRNA 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 mecha-nism.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 signifi-cantly 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 in-creased 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 in-creased 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 molecular mechanism is related to LncRNA MALAT1 regulating the expression of H3K27me3 by recruiting EZH2.
高飞;谈园;张莲;罗立芳;罗雯;马乐;黎姣艳;黄敖
湖南省长沙市第一医院,湖南 长沙 410005湖南省中医学院第一附属医院皮肤科,湖南 长沙 410005
甲基化转移酶表观遗传学沉默系统性红斑狼疮抗炎因子
MethyltransferaseEpigenetic silencingSystemic lupus erythematosusAnti-in-flammatory factor
《河北医学》 2024 (009)
1434-1439 / 6
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)
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