酵母单杂交筛选人食管癌细胞系SHEEC中NF-kappa B元件结合蛋白  

Screening NF-kappa B Element Binding Protein of Esophageal Carcinoma Cell Line SHEEC With Yeast One Hybrid System

作　　者：牛永东[1] 许丽艳[2] 韩溟[1] 方王楷[1] 沈忠英[2] 李恩民[1] 

机构地区：[1]汕头大学医学院生化与分子生物学教研室 [2]汕头大学医学院肿瘤病理研究室

出　　处：《癌变．畸变．突变》2006年第4期310-313,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家自然科学基金面上项目(No.30170428;No.30370641;No.30570849);广东省自然科学基金重点项目(No.37788;No.05104541);教育部博士学科科研基金项目(No.20020560002;No.20050560003)

摘　　要：背景与目的:从食管癌细胞cDNA文库中筛选能够特异性结合NF-kappaB核心元件的蛋白质分子,探讨NGAL基因的表达调控机制,探寻NGAL基因在食管上皮细胞癌变中过表达的原因。材料与方法:应用酵母单杂交系统,以三重NF-kappaB元件寡聚核苷酸串为诱饵,对人食管癌细胞系SHEECcDNA酵母杂交文库进行筛选,应用DNA测序及生物信息学技术对筛选克隆进行分析。结果:共获得30个阳性克隆,包括4种编码蛋白,2个hypotheticalprotein,2种线粒体基因,3类非编码RNA等。结论:应用酵母单杂交手段筛选出食管癌细胞NF-kappaB核心元件结合蛋白,但只是一种初步的实验,还需进一步研究证实。BACKGROUND＆AIM： This study was to screen an esophageal cancer cell line （SHEEC） cDNA library for the cDNA that expressed a protein that can specifically bind the NF-kappa B element, for further understanding the expression and regulation mechanism of the NGAL（neutrophil gelatinase-associated lipocalin） gene, and the course for the over-expression of the NGAL gene in the esophageal epithelial squamous carcinoma. MATERAL AND METHODS： A tandem three oligo-nucleotides of NF-kappa B was employed as a bait in a yeast one hybrid system to screen the eDNA library of the human esophageal carcinoma cell line （SHEEC） The eDNA clones obtained were sequenced and analyzed with bioinformatics tools. RESULTS： Among the thirty clones obtained, four types of clones could encode proteins, two clones were hypothetical proteins, two types of clones were mitochondrial genes, and three types of clones were non-encoding sequences. CONCLUSION： Using the yeast one hybrid system to screen the eDNA library for the NF-kappa B core element is a preliminary work that needs confirmation by further study.

关 键 词：酵母单杂交 食管癌 CDNA文库 

分 类 号：R73-3[医药卫生—肿瘤]