LncRNA FOXM1-AS靶向miR-499a-5p抑制香烟诱导的人肺上皮细胞炎症因子表达和上皮间质转化  

LncRNA FOXM1-AS targets miR-499a-5p to suppress cigarette smoke-induced expression of inflammatory factors and EMT in BEAS-2B cells

作　　者：黄佳茹 袁亚平[1] 王林宣 刘源 杨俊侠 郭晓霞 邓国平[1] 顾文超[1] Huang Jiaru;Yuan Yaping;Wang Linxuan;Liu Yuan;Yang Junxia;Guo Xiaoxia;Deng Guoping;Gu Wenchao(Department of Respiratory and Critical Care Medicine,Shanghai Pudong New Area People′s Hospital,Shanghai 201200,China)

机构地区：[1]上海市浦东新区人民医院呼吸与危重症医学科,上海201200

出　　处：《国际呼吸杂志》2023年第6期664-669,共6页International Journal of Respiration

基　　金：浦东新区卫生健康委员会临床特色学科建设资助项目(PWYts2021-18)。

摘　　要：目的探讨叉头蛋白M1反义长链非编码RNA(LncRNA FOXM1-AS)对香烟烟雾提取物(CSE)诱导的支气管上皮细胞(BEAS-2B)炎症因子表达和上皮-间质转化(EMT)的影响, 并研究其潜在机制。方法本研究为实验研究。培养BEAS-2B, 采用随机数字表法分成对照组, CSE组(2.5% CSE处理)、siNC+CSE组(转染siNC, 给予2.5% CSE处理)、siFOXM1-AS+CSE组(转染siFOXM1-AS, 给予2.5% CSE处理);通过实时荧光定量多聚核苷酸链式反应方法、酶联免疫吸附试验法和蛋白质印迹法检测各组FOXM1-AS、IL-6、TNF-α、E-cadherin、Vimentin基因表达水平和IL-6、TNF-α水平以及SOX6蛋白表达;利用生物信息学软件miRcode预测FOXM1-AS的潜在靶基因(miR-499a-5p), 荧光素酶报告基因实验鉴定二者的靶向关系。结果与对照组比较, CSE组细胞中FOXM1-AS的表达水平显著增加(1.06±0.08比2.89±0.31, P<0.01)。利用siFOXM1-AS沉默FOXM1-AS后, 明显抑制了CSE诱导的细胞炎症因子IL-6(2.63±0.34比1.39±0.23, P<0.01)、TNF-α(2.58±0.23比1.35±0.20, P<0.01)的表达, 细胞EMT进程中, E-cadherin基因表达水平增加(0.35±0.10比0.92±0.08, P<0.01), 而Vimentin基因表达水平减少(2.37±0.28比1.41±0.20, P<0.01)。此外, FOXM1-AS通过靶向结合miR-499a-5p调控SOX6表达。miR-499a-5p抑制剂可逆转siFOXM1-AS对CSE诱导的细胞炎症因子表达及EMT的作用。结论下调FOXM1-AS靶向miR-499a-5p可抑制CSE诱导的BEAS-2B细胞炎症因子表达和EMT。Objective To investigate the effects of long non-coding RNA antisense to FOXM1(LncRNA FOXM1-AS)on cigarette smoke extract(CSE)-induced expression of inflammatory factors and epithelial-mesenchymal transition(EMT)in human lung epithelial cells BEAS-2B and to explore the mechanisms.Methods This was an experimental study.BEAS-2B was cultured and randomly assigned into 4 groups by random number table method,including the control group,CSE group(2.5%CSE treatment),siNC+CSE group(siNC transfection and 2.5%CSE treatment),and siFOXM1-AS+CSE group(siFOXM1-AS transfection and 2.5%CSE treatment).qPCR,ELISA assay,and Western blotting were used to examine the expression of the mRNA level of FOXM1-AS,IL-6,TNF-α,E-cadherin,and Vimentin and the expression of IL-6,TNF-α,and SOX6.Bioinformatics software miRcode was used to predict the potential target miRNAs miR-499a-5p of FOXM1-AS,and then luciferase reporter assay was used to verify the relationship between them.Results Compared with the control group,the expression level of FOXM1-AS was significantly increased in CSE group(1.06±0.08 vs 2.89±0.31,P<0.01).FOXM1-AS silencing with siFOXM1-AS significantly inhibited the expression of CSE-induced inflammatory cytokines IL-6(2.63±0.34 vs 1.39±0.23,P<0.01)and TNF-α(2.58±0.23 vs 1.35±0.20,P<0.01),and FOXM1-AS silencing with siFOXM1-AS significantly delayed the CSE-induced EMT process.The expression level of E-cadherin gene was increased(0.35±0.10 vs 0.92±0.08,P<0.01),but the expression level of Vimentin gene was decreased(2.37±0.28 vs 1.41±0.20,P<0.01).Furthermore,FOXM1-AS modulates the expression of SOX6 by targeting miR-499a-5p.MiR-499a-5p inhibitor can reverse the effect of siFOXM1-AS on CSE-induced inflammatory cytokine expression and EMT.Conclusions Suppressed FOXM1-AS targeting miR-499a-5p can inhibit CSE-induced inflammatory cytokine expression and EMT in BEAS-2B cells.

关 键 词：炎症 上皮-间质转化 长链非编码RNA FOXM1-AS 微小RNA499a-5p 

分 类 号：R563[医药卫生—呼吸系统]