组蛋白乙酰化对A549和BEAS-2B细胞哮喘易感基因ADAM33的影响  

Effect of histone acetylation on asthma susceptibility gene ADAM33 in A549 and BEAS-2B cells

作　　者：郝忠分 杜娟[1] 芮菲菲 杭芸 汤陈璐 李章 束进 HAO Zhongfen;DU Juan;RUI Feifei;HANG Yun;TANG Chenlu;LI Zhang;SHU Jin(Department of Pediatrics,the Fourth Affiliated Hospital of Jiangsu University,Zhenjiang Jiangsu 212001;Department of Neonatal,Changzhou Maternal and Child Health Hospital,Changzhou Jiangsu 213000,China)

机构地区：[1]江苏大学第四附属医院儿科,江苏镇江212001 [2]常州市妇幼保健院新生儿科,江苏常州213000

出　　处：《江苏大学学报（医学版）》2024年第3期242-247,253,共7页Journal of Jiangsu University：Medicine Edition

基　　金：镇江市重点研发计划社会发展项目(SH2018078)。

摘　　要：目的:研究丁酸钠、曲古抑菌素A(trichostatin A,TSA)及腺病毒E1A相关的300 kD蛋白(adenoviral E1A binding protein of 300 kD,P300)介导的组蛋白乙酰化在人非小细胞肺癌A549细胞及人支气管上皮BEAS-2B细胞中对哮喘易感的解整合素金属蛋白酶33(a disintegrin and metalloproteinase 33,ADAM33)基因表达的影响。方法:将A549细胞及BEAS-2B细胞分别分为丁酸钠对照组(双蒸水处理)和1、2.5、5 mmol/L丁酸钠组,TSA对照组(0.1%二甲基亚砜处理)和0.2、0.4、0.8μmol/L TSA组,按照组别分别予以相应处理;另将BEAS-2B细胞分为对照组(转染P300突变质粒)和P300组(转染P300表达质粒);采用双荧光素酶报告基因法分析ADAM33启动子活性的变化,实时荧光定量PCR(quantitative real time-PCR,qRT-PCR)检测ADAM33 mRNA表达,蛋白质印迹法检测ADAM33蛋白表达。结果:在人非小细胞肺癌A549细胞中,与对照组相比,1 mmol/L丁酸钠组及0.2μmol/L TSA组ADAM33基因启动子活性明显降低(P<0.01);在BEAS-2B细胞中,与对照组相比,1 mmol/L丁酸钠组及0.2μmol/L TSA组ADAM33基因启动子活性、mRNA及蛋白相对表达水平明显降低(P<0.05或P<0.01)。加大丁酸钠、TSA药物浓度,ADAM33表达无显著差异。在人支气管上皮BEAS-2B细胞中,与对照组相比,P300组ADAM33启动子活性、mRNA和蛋白相对表达水平明显降低(P<0.05)。结论:丁酸钠、TSA通过组蛋白乙酰化降低人非小细胞肺癌A549细胞和人支气管上皮BEAS-2B细胞中ADAM33表达,P300通过组蛋白乙酰化降低人支气管上皮BEAS-2B细胞中ADAM33表达。Objective:To evaluate the effects of histone acetylation mediated by sodium butyrate,trichostatin A(TSA)and adenoviral E1A binding protein of 300 kD(P300)on the expression of a disintegrin and metalloproteinase 33(ADAM33),one of asthma susceptible genes,in human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells.Methods:Human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells were divided into sodium butyrate control group(double distilled water treatment)and 1,2.5,5 mmol/L sodium butyrate groups,TSA control group(0.1%dimethyl sulfoxide treatment)and 0.2,0.4,0.8μmol/L TSA groups,respectively.BEAS-2B cells were divided into P300 control group(transfected with P300 mutant plasmid)and P300 group(transfected with P300 expressing plasmid).These cells were treated respectively according to the groups.The ADAM33 promoter activity was analyzed by dual luciferase reporter method,the expression of ADAM33 mRNA was detected by quantitative real time-PCR(qRT-PCR),and the expression of ADAM33 protein was detected by Western blotting.Results:In human non-small cell lung cancer A549 cells,compared with the control group,ADAM33 promoter activity was significantly decreased in 1 mmol/L sodium butyrate group and 0.2μmol/L TSA group(P<0.01).In human bronchial epithelial BEAS-2B cells,compared with the control group,the activity of ADAM33 gene promoter,mRNA and protein expression levels was greatly decreased in 1 mmol/L sodium butyrate group and 0.2μmol/L TSA group(P<0.05 or P<0.01).There was no significant difference in ADAM33 expression when increasing the concentration of sodium butyrate and TSA.In BEAS-2B cells,compared with the control group,the promoter activity,mRNA and protein expression levels of ADAM33 in P300 group were markedly decreased(P<0.05).Conclusion:Sodium butyrate and TSA decreased ADAM33 expression in human non-small cell lung cancer A549 cells and human bronchial epithelial BEAS-2B cells through histone acetylation,while P300 decreased ADAM33 exp

关 键 词：哮喘 解整合素金属蛋白酶33 丁酸钠 曲古抑菌素A 人支气管上皮BEAS-2B细胞 

分 类 号：R725.6[医药卫生—儿科]