食管鳞癌细胞中NF-κB与IκBα的mRNA、蛋白表达  

Nuclear Factor-κB and IκBα mRNA,Protein Overexpression in Esophageal Squamous Cell Carcinoma Cell Lines

作　　者：田芳[1] 侯卫红[1] 许培荣[1] 王建民[1] 陈华艳[1] 薛乐勋[1] 陈奎生[2] 

机构地区：[1]郑州大学细胞生物学研究室,郑州市450052 [2]郑州大学第一附属医院病理科

出　　处：《中国肿瘤临床》2006年第4期198-201,共4页Chinese Journal of Clinical Oncology

基　　金：教育部"十五"211工程重点建设项目资助(编号:教重办[2002]第2号)

摘　　要：目的:核转录因子NF-kappaB(NF-κB)是一种重要的转录因子,参与调控多种与炎症、抗凋亡、肿瘤形成和转化有关的基因表达。本研究通过检测食管鳞癌细胞中NF-κB与IκBαmRNA、蛋白的表达及NF-κB的DNA结合活性,分析NF-κB在食管鳞癌细胞中的激活状态及其在食管鳞癌细胞的生存及转化中的作用。方法:采用West-ernblotting法检测两株食管鳞癌细胞胞质中NF-κB亚单位p50和p65、阻遏物IκBα及其磷酸化IκBα的蛋白表达,检测细胞核中p50和p65的蛋白表达并采用EMSA法检测其与DNA的结合活性。使用RT-PCR法检测p50、p65、IκBαmRNA的表达。结果:NF-κB的亚单位p50、p65、IκBα及磷酸化IκBα在食管鳞癌细胞质中均表达,p50、p65蛋白在细胞核中也表达并具有较高的DNA结合活性。RT-PCR结果表明,p50、p65、IκBα的mRNA在细胞中也存在过表达。结论:本研究发现NF-κB在两株食管鳞癌细胞系中被激活,基因的过表达及IκBα的磷酸化在维持NF-κB的活化中起着重要的作用。Objective： To investigate mRNA and protein expression levels of NF-κB and IκBα, DNA-binding activity of NF-κB in ESCC cell lines. Methods： Western blotting was used to determine the protein levels of the subunits of NF-κB, p50 and p65, inhibitor-κBα and p-IκBα in two ESCC cell lines Eca109 and EC9706. Nuclear proteins from the ESCC cell lines were extracted to evaluate the protein levels of p50, p65 and the DNA-binding activity by electrophoretic mobility gel shift assay （EMSA）. RT-PCR was used to detect the mRNA of p50, p65 and IκBα. Results： ESCC cell lines constitutively overexpressed p50, p65, IκBα and p- IκBα in the cytoplasm. The proteins of p50 and p65 were also expressed in the nuclei and had high DNA-binding activity. In addition, the results of RT-PCR showed overexpression of p50, p65 and IκBα mRNA. Conclusion：The findings indicate that NF-κB is constitutively activated in human ESCC cell lines, which may be caused by its gene overexpression or the phosphorylation of IκBα, suggesting that NF-κB may play a critical role in carcinogenesis of the esophagus.

关 键 词：食管鳞癌 NF—κB IΚBΑ 

分 类 号：R735.1[医药卫生—肿瘤]