LncRNA MALAT1/miR-876-5p/FOXM1轴对TNF-α诱导的HaCaT细胞增殖、凋亡和炎症反应的影响  

Influences of LncRNA MALAT1/miR-876-5p/FOXM1 axis on TNF-α-induced proliferation,apoptosis and inflammatory response of HaCaT cells

作　　者：金曌 刘钟[1] 彭静 胡荣毅[1] 吴娟[1] 黄琴斯[1] 王飞 JIN Zhao;LIU Zhong;PENG Jing;HU Rongyi;WU Juan;HUANG Qinsi;WANG Fei(Department of Dermatology,Wuhan First Hospital,Wuhan 430022,China;School of Acupuncture and Bone Injury,Hubei University of Traditional Chinese Medicine,Wuhan 430022,China)

机构地区：[1]武汉市第一医院皮肤科,武汉430022 [2]湖北中医药大学针灸骨伤学院,武汉430022

出　　处：《中国免疫学杂志》2025年第3期582-588,594,共8页Chinese Journal of Immunology

基　　金：武汉市卫生健康委科研项目(WX21A13)。

摘　　要：目的:探讨长链非编码RNA肺腺癌转移相关转录子1(lncRNA MALAT1)/miR-876-5p/叉头框蛋白质M1(FOXM1)轴对TNF-α诱导的HaCaT细胞增殖、凋亡和炎症反应的影响。方法:将HaCaT细胞分为Ct组、Model组、si-NC组、si-MALAT1组、mimic NC组、miR-876-5p mimic组、si-MALAT1+inhibitor NC组、si-MALAT1+miR-876-5p inhibitor组,除Ct组外,其余组细胞均需用25μg/L TNF-α处理以诱导银屑病体外细胞模型,TNF-α诱导24 h后再转染对应的转染物48 h,用于后续实验。qRT-PCR检测细胞中MALAT1、miR-876-5p表达;CCK-8、EdU染色检测细胞增殖;流式细胞术检测细胞凋亡;ELISA检测细胞上清液中IL-6、TNF-α、IL-1β水平;Western blot检测FOXM1、增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)蛋白表达;双荧光素酶报告基因实验验证MALAT1与miR-876-5p、miR-876-5p与FOXM1的关系;RNA pull down实验验证MALAT1与miR-876-5p的关系。结果:相较于对照组,实验组MALAT1、FOXM1蛋白表达增加,miR-876-5p表达下降(P<0.05);相较于Ct组,Model组HaCaT细胞中MALAT1表达、FOXM1蛋白表达、A450值、EdU阳性率、细胞上清液中IL-6、TNF-α、IL-1β水平、PCNA、Bcl-2蛋白表达升高,miR-876-5p表达、细胞凋亡率、Bax蛋白表达降低(P<0.05);沉默MALAT1或过表达miR-876-5p可抑制TNF-α诱导的HaCaT细胞增殖、炎症反应,促进细胞凋亡;miR-876-5p inhibitor恢复了MALAT1敲低对TNF-α刺激的HaCaT细胞凋亡、增殖及炎症反应的作用;MALAT1靶向下调miR-876-5p,miR-876-5p靶向下调FOXM1。结论:敲低MALAT1可能通过提高miR-876-5p表达来下调FOXM1表达,进而促进TNF-α刺激的HaCaT细胞凋亡,减轻炎症反应并降低增殖。Objective:To investigate the influences of long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1(lncRNA MALAT1)/miR-876-5p/forkhead box protein M1(FOXM1)axis on TNF-α-induced proliferation,apoptosis and inflammatory response of HaCaT cells.Methods:HaCaT cells were grouped into Ct group,Model group,si-NC group,si-MALAT1 group,mimic NC group,miR-876-5p mimic group,si-MALAT1+inhibitor NC group,and si-MALAT1+miR-876-5p inhibitor group.Except for the Ct group,cells in other groups were treated with 25μg/L TNF-α to induce the in vitro cell model of psoriasis,and after 24 hours of TNF-α induction,the corresponding transfectants were transfected for 48 hours for subsequent experiments.qRT-PCR was applied to detect the expression of MALAT1 and miR-876-5p in cells;CCK-8 method and EdU staining were applied to detect cell proliferation;flow cytometry was applied to detect apoptosis;ELISA method was applied to detect the levels of IL-6,TNF-α,and IL-1β in cell supernatant;Western blot was applied to detect the protein expressions of FOXM1,proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax)and B-lymphocytoma-2(Bcl-2);dual-luciferase reporter gene assay was applied to verify the relationship between MALAT1 and miR-876-5p,miR-876-5p and FOXM1;and RNA pull down experiments were applied to verify the relationship between MALAT1 and miR-876-5p.Results:Compared with the control group,the expressions of MALAT1 and FOXM1 protein expression in the experimental group were increased,and the expression of miR-876-5p was decreased(P<0.05);compared with Ct group,the expressions of MALAT1 and FOXM1 protein expression in HaCaT cells,A450 value,EdU positive rate,the levels of IL-6,TNF-α,IL-1β,and the protein expressions of PCNA and Bcl-2 in cell supernatant in Model group increased,the expression of miR-876-5p,apoptosis rate and the protein expression of Bax were decreased(P<0.05);silencing MALAT1 or overexpressing miR-876-5p could inhibit the proliferation and inflammatory response of HaCaT ce

关 键 词：肺腺癌转移相关转录子1 miR-876-5p 叉头框蛋白质M1 细胞增殖 银屑病 

分 类 号：R275.9[医药卫生—中西医结合]