miR-542-3p对脂多糖诱导的人支气管上皮细胞增殖和迁移的影响及机制研究  

Effect and mechanism of miR-542-3p on the proliferation and migration of LPS-induced human bronchial epithelial cells

作　　者：石晓岚[1] 赵龙[1] 王宁[1] 刘翠翠[1] 王静[1] 马彩铃[1] SHI Xiaolan;ZHAO Long;WANG Ning;LIU Cuicui;WANG Jing;MA Cailing(Respiratory Asthma Center,Xi’an Children’s Hospital,Xi’an 710002,China)

机构地区：[1]西安市儿童医院呼吸哮喘中心,陕西西安710002

出　　处：《陕西医学杂志》2021年第10期1218-1222,共5页Shaanxi Medical Journal

基　　金：西安市儿童医院院级科研项目(2018C04)。

摘　　要：目的:探讨miR-542-3p在儿童呼吸哮喘中的作用及其可能机制。方法:用不同浓度的脂多糖(LPS)分别处理BEAS-2B细胞24 h构建哮喘细胞模型,采用脂质体法将miR-542-3p模拟物、抑制物及对照转染至BEAS-2B。RT-qPCR检测miR-542-3p的表达水平,CCK-8和划痕法检测细胞增殖和迁移能力。RT-qPCR和western blot法检测白细胞介素33(IL-33)和可溶性生长刺激表达基因2蛋白(sST2)的mRNA和蛋白表达水平的变化。通过在线网站microRNA.org预测分析miR-542-3p与IL-33的靶向关系,使用双萤光素酶报告基因实验进行验证,并对miR-542-3p与IL-33进行Spearman相关性分析。通过共转染miR-542-3p和pcDNA3.1-IL-33,检测IL-33对miR-542-3p功能的影响。结果:与未处理组(untreated)相比,miR-542-3p在LPS处理的BEAS-2B细胞中相对表达量显著下调(P<0.05),而IL-33和sST2的mRNA相对表达量均显著上调(均P<0.05)。miR-542-3p下调IL-33的表达(P<0.05),过表达IL-33抑制miR-542-3p对LPS诱导的BEAS-2B增殖和迁移的影响(均P<0.05)。结论:miR-542-3p可能通过抑制IL-33/sST2信号通路缓解儿童呼吸哮喘。Objective:To investigate the effect of miR-542-3p on childhood respiratory asthma and its mechanism.Methods:BEAS-2B cells were treated with different concentrations of Lipopolysaccharide(LPS)for 24 hours to construct asthma cell model.The miR-542-3p mimic,inhibitor as well as their respective controls were transfected into BEAS-2B via liposome method.RT-qPCR was used to detect the expression level of miR-542-3p.CCK-8 and wound-healing assay were used to determine the effect of miR-542-3p on the cell proliferation and migration.The mRNA and protein expression levels of IL-33 and sST2 were detected by RT-qPCR and Western blot.The relationship between miR-542-3p and IL-33 was predicted by microRNA.org and verified by double-luciferase reporter gene experiment.Spearman correlation analysis was performed to analyze the correlation between miR-542-3p and IL-33.The miR-542-3p and pcDNA3.1-IL-33 were co-transfected to detect the effect of IL-33 on the function of miR-542-3p.Results:Compared with untreated group,the relative mRNA expression of miR-542-3p was significantly inhibited while IL-33 and sST2 were up-regulated in LPS-stimulated BEAS-2B(all P<0.05).The miR-542-3p down-regulated the expression of IL-33,while overexpressing IL-33 inhibited the effect of miR-542-3p on LPS-induced BEAS-2B proliferation and migration(P<0.05).Conclusion:In LPS-induced BEAS-2B,miR-542-3p may alleviate childhood respiratory asthma by inhibiting IL-33/sST2 signaling pathway.

关 键 词：miR-542-3p IL-33/sST2信号通路 儿童呼吸哮喘 细胞增殖 细胞迁移 

分 类 号：R392.12[医药卫生—免疫学]