双向电泳和质谱技术研究鼻咽癌候选抑瘤基因NGX6的功能  

Preliminary Function Study of a Putative Tumor Suppressor Gene in Human Nasopharyngeal Carcinoma Cells Using Two-Dimensional Electrophoresis and Mass Spectrometry

作　　者：李江[1] 谭琛[1] 向秋[1] 张小梅[1] 马健[1] 李伟芳[1] 王洁如[1] 梁宋平[2] 李桂源[1] 

机构地区：[1]中南大学湘雅医学院肿瘤研究所细胞遗传室,湖南长沙410078 [2]湖南师范大学生命科学院,湖南长沙410006

出　　处：《癌症》2001年第7期692-695,共4页Chinese Journal of Cancer

基　　金：国家"863"计划(No. 102-10-01-05);国家自然科学基金项目(No.39900163)

摘　　要：目的：NGX6基因是我室克隆的与鼻咽癌相关的肿瘤抑制候选基因,它的功能与作用机制目前尚不清楚。方法：通过亚克隆的方法将NGX6 cDNA 片段克隆至pcDNA3.1(＋)的表达载体，经脂质体导入HNE1细胞,采用双向凝胶电泳分离细胞内蛋白质，筛选差异表达的蛋白质点进行质谱分析和生物信息学资料处理。结果：我们鉴定出7种表达下调的蛋白质，其中包括RAB蛋白、NG24前体、LIM同源盒9等。这些蛋白参与了细胞生长、增殖和分化的信号传导通路。结论：这些结果说明NGX6基因可能通过多种途径影响鼻咽癌的生长。本文为研究NGX6的作用机制提供了很好的实验资料，对鼻咽癌的基因治疗奠定了一定的研究基础，也为研究其他基因的作用机制提供了新思路。Objective：This study was designed to reveal the preliminary function of a novel candidate for tumor suppressor gene, named NGX6, to nasopharyngeal carcinoma (NPC) cells. Method：NGX6 was introduced into HNE1 cells by liposome transfection. The authors used two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) to identify proteins that were differently expressed in NGX6 transfected cells. After staining and image analysis, spots of interest were isolated and subjected to mass spectrometry (MS). Results：We identified seven proteins, which were down-regulated in NGX6 transfected cells. These proteins included RAB protein, NG24 precusor, and LIM-homebox 9; which involved in cell cycling, transcription regulation, and metabolism. Conclusion：NGX6 may exert its effect in NPC cells by regulating the expression of these proteins.

关 键 词：双向电泳 基质辅助性 激光解吸飞行时间质谱 细胞转染 NGX6基因 鼻咽癌 

分 类 号：R739.63[医药卫生—肿瘤] 756[医药卫生—临床医学]