HMGB1对HTLV-1病毒Tax蛋白表达的T细胞中NF-κB活性的影响  

The effect of HMGB1 on the activity of NF-κB in T lymphocytes expressing HTLV-1 Tax protein

作　　者：刘苏慧[1] 牛志国[1] 李俊英[1] 胡新攀[1] 赵玲玲[1] 范勇利[1] 陈丽媛[1] 王辉[1] 

机构地区：[1]新乡医学院医学检验系,新乡453003

出　　处：《中国免疫学杂志》2010年第11期973-976,981,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.30972755)资助

摘　　要：目的:构建人高迁移率组蛋白1(high mobility group box-1,HMGB1)的真核表达载体,转入TaxP及TaxN细胞进行表达,并研究其对Tax蛋白表达的T细胞中NF-κB活性的影响。方法:用RT-PCR方法扩增人T淋巴细胞系Jurkat细胞中HMGB1的cDNA,连接于真核表达载体pcDNA3.0;将pcDNA3.0-HMGB1转染TaxP及TaxN细胞,48小时后检测RT-PCR检测HMGB1和Tax mRNA的表达,HMGB1及p65蛋白的表达;将pcDNA3.0-HMGB1和NF-κB报告基因共转染TaxP及TaxN细胞48小时后检测荧光素酶活性。结果:成功构建了重组表达质粒pcDNA3.0-HMGB1;Tax可以促进HMGB1mRNA和蛋白的表达,HMGB1可以促进p65mRNA和蛋白的表达,并能上调NF-κB活性。结论:HMGB1能协同Tax蛋白激活NF-kB。Objective：To construct eukaryotic expressing plasmid pcDNA3.0-hHMGB1 and transiently express it in TaxP and TaxN cells for researching the effect of HMGB1 on activity of NF-κB.Methods：Total mRNA was extracted from Jurkat cell,and hHMGB1 cDNA was amplified by RT-PCR,then it was inserted into a eukaryotic expressing plasmid pcDNA3.0 to construct pcDNA3.0-hHMGB1.The constructed plasmid was transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method,and the expression levels of HMGB1,p65 and Tax mRNA in transfected cells were assayed by using RT-PCR after 48 hours.Meanwhile,the proteins of HMGB1 and p65 were detected by Western blot.The constructed plasmid and pNF-κB-luc reporter gene plasmid were co-transfected into TaxN and TaxP cells,and the activity of luciferase was assayed after 48 hours.Results：The results showed that eukaryotic expressing plasmid pcDNA3.0-hHMGB1 was successfully constructed.The mRNA and protein expression of HMGB1 could be promoted significantly by Tax.HMGB1 can promote the mRNA and protein expressions of p65 in TaxN and TaxP cells,the activity of NF-κB was up-regulated by HMGB1 either.Conclusion：HMGB1 could activate NF-κB pathway by synergy with HTLV-1 Tax protein.

关 键 词：高迁移率组蛋白 成人T细胞白血病病毒1 NF-ΚB 

分 类 号：R392[医药卫生—免疫学]