JNK3抑制NF-κB转录活性的研究  

JNK3 Inhibits the Transcriptional Activity of NF-κB

作　　者：韩卿[1] 宋方洲[1] 易发平[1] 卜友泉[1] 李长燕[2] 杨晓明[2] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室临床检验诊断学省部共建教育部重点实验室,重庆市400016 [2]军事医学科学院放射与辐射医学研究所,北京市100850

出　　处：《医学分子生物学杂志》2008年第4期287-291,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金(No.30671008)~~

摘　　要：目的研究人c-Jun氨基末端激酶3(c-Jun N-terminal kinases3,JNK3)对核因子κB(nuclear factor kappa B,NF-κB)转录活性的影响。方法构建人JNK3真核表达载体pcDNA3.1-JNK3,与3×κB-luc报告基因质粒共转染人胚胎肾细胞293(HEK293),分别进行如下处理:共转染50ngpCMV-Myc-p65质粒;转染24h后,加入浓度为10ng/ml的TNF-α或IL-1β刺激6h,收集细胞,检测荧光素酶活性。结果成功构建了pcDNA3.1-JNK3真核表达载体;提高细胞内p65水平,加入TNF-α或IL-1β刺激均可明显激活NF-κB的转录,JNK3对NF-κB的转录活性有明显抑制作用,且随着JNK3转染剂量的增加,抑制作用增强。结论JNK3能明显抑制NF-κB的转录活性。Objective Research the effect of JNK3 on the transcriptional activity of NF-κB. Methods The JNK3 eukaryotic expression vector pcDNA3. 1-JNK3 was constructed as described and co-transfected with 3 x KB-luc plasmid into HEK293 cells. Subsequently, the cells were either transfected with pCMV-Myc-p65 plasmid or 24h after transfection treated with 10 ng/ml of IL-1 β/TNF-α for 6 h, and then harvested for luciferase activity analysis. Results The constructed eu- karyotic expression vector pcDNA3. 1-JNK3 expressed high level of p65 protein in transfected cells. Addition of TNF-α or IL-1β to the cells significantly activated the NF-KB reporter gene, whereas JNK3 clearly inhibited the transcriptional activity of the activated NF-κB. With the increase of the transfection dose of JNK3, the inhibitive effect was enhanced. Conclusion JNK3 inhibits the transcriptional activity of NF-κB.

关 键 词：JNK3 核因子ΚB 荧光素酶报告基因 转录活性 

分 类 号：Q257[生物学—细胞生物学]