不同转移潜能人肝癌细胞系酪氨酸磷酸化蛋白质差异分析  

Differential analysis of tyrosine-phosphorylated proteins in human hepatocellular carcinoma cell lines with different metastasis potentials

作　　者：张丽君[1] 崔杰峰[1] 刘银坤[1] 朱运松[2] 宋海燕[1] 申华莉[3] 冯巨涛[1] 代智[1] 陈洁[1] 孙瑞霞[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所,上海200032 [2]复旦大学海医学院分子遗传教研室 [3]复旦大学蛋白质组学研究中心

出　　处：《中华肝脏病杂志》2005年第6期436-439,共4页Chinese Journal of Hepatology

基　　金：国家重点基础研究(973)项目(2001CB510205)国家自然科学基金(3017416)

摘　　要：目的研究不同转移潜能人肝癌细胞系中酪氨酸磷酸化蛋白质表达的差异并筛查与肝癌转移相关的酪氨酸磷酸化蛋白质分子。方法应用双电泳(2-D E)、免疫印迹法及基质辅助激光解析电离飞行时间质谱分析,对三种不同转移潜能人肝癌细胞系Hep 3B、MHCC97L和MHCC97H进行酪氨酸磷酸化蛋白质组分析。结果对照2-DE胶和免疫印迹发光胶片,Hep3B检测到10个点,MHCC 9 7L检测到19个点, MHCC97H检测到17个点。经质谱鉴定,得到膜连蛋白Ⅰ等19个差异点。结论细胞内酪氨酸磷酸化蛋白的表达差异与肝癌的侵袭转移有关。Objectives To compare expressions of tyrosine-phosphorylated proteins in different hepatocel-lular carcinoma cell lines with different metastasis potential and to screen key molecules associated with HCC metastasis and recurrence. Methods Using two-dimensional electrophoresis, Western blotting and MALDI-TOF-MS/MS, we analyzed tyrosine-phosphorylated protein profiles of Hep3B, MHCC97L and MHCC97H, HCC cell lines with different metastasis potentials. Results 10 spots were detected in Hep3B, 19 in MHCC97L and 17 in MHCC97H. Seventeen significantly different phosphotyrosine proteins in gel were identified by MALDI-TOF-MS/MS, including Annexin I. Conclusion The changed expression of tyrosine-phosphorylated proteins is associated with HCC metastasis and recurrence.

关 键 词：人肝癌细胞系 酪氨酸磷酸化蛋白质 转移潜能 差异分析 HEP3B 蛋白质组分析 蛋白质分子 蛋白质表达 免疫印迹法 肝癌转移 方法应用 质谱分析 飞行时间 激光解析 基质辅助 2-DE 质谱鉴定 侵袭转移 表达差异 检测 蛋白Ⅰ 细胞内 

分 类 号：R735.7[医药卫生—肿瘤] R543.2[医药卫生—临床医学]