肝癌亚细胞结构的蛋白质组分比较分析  

COMPARATIVE PROTEOME ANALYSIS OF HEPATOMA CELLS AT SUBCELLULAR LEVEL

作　　者：李兴[1] 潘卫[2] 邱峰[3] 邱宗荫[1] 

机构地区：[1]重庆医科大学医学检验系,重庆400016 [2]贵阳医学院医学检验系,贵阳550004 [3]重庆医科大学药学系,重庆400016

出　　处：《分子细胞生物学报》2006年第5期399-406,共8页Journal of Molecular Cell Biology

基　　金：国家自然科学基金(30476021);教育部博士点基金(教特发中心函[2004]166号)资助项目

摘　　要：运用亚细胞蛋白质组学的研究策略,分离纯化亚细胞结构,可以提高低丰度蛋白质在双向电泳中检出的数量。通过对比分析肝癌细胞与正常肝细胞线粒体、细胞核蛋白质组的差异表达情况,为肝癌发病机理的研究提供更多、更有价值的信息。以体外培养的人体肝癌细胞QGY-7703与正常肝细胞LO2为研究模型,通过超离心的方法分离细胞的线粒体和细胞核。双向电泳分离线粒体和细胞核的蛋白质,图像分析筛选差异表达蛋白斑点,MALDI-TOF-MS鉴定蛋白质。从线粒体、细胞核的蛋白质电泳图谱中筛选出54个候选差异表达的蛋白质斑点,质谱鉴定出22种差异表达蛋白质,其中17种在肝癌细胞中表达上调,5种在肝癌细胞中表达下调。筛选出的差异表达蛋白质涉及到细胞的能量代谢、蛋白质合成、细胞骨架与核骨架的改变、mRNA的加工成熟及凋亡调控等许多方面,表明癌变细胞的组织结构和代谢状态都发生了很大的变化。the subcellular proteome strategy can complement the separation power of two-dimensional electrophoresis,a step for subcellular fractionation is introduced before electrophoresis is conducted,and more proteins will be displayed in two-dimensional gels. A comparative analysis of proteomic profiling of mitochondria and nuclei was conducted between hepatoma cell and hepatocyte,in order to find more information involved in cancer development. The cultured hepatoma cell line QGY-7703 and hepatocyte line LO2 were used as research models in this work. Subcellular fractionation for mitochondrion and nucleus were done by uhracentrifugation,then two-dimensional electrophoresis was applied to the separation of mitochondrial and nuclear proteins,imaging analysis for the selection of differentially-expressed protein spots,and MALDI-TOF-MS for the identification of proteins. 54 spots from electrophoresis gels were selected as differentially-expressed protein for MS analysis which resulted in identification for 22 proteins. Among the 22 differentiallyexpressed proteins,17 show a up-regulated expression and 5 show a down-regulated expression. These differentially-expressed proteins found in this work have a wide coverage of functions which are related to energy metabolism,cytosheleton,protein biosynthesis,pre-mRNA splicing and processing,apoptosis regulation. These results imply that cancer cell has experienced a fundamental change in structure and metabolism pattern.

关 键 词：肝癌细胞 亚细胞蛋白质组 线粒体 细胞核 

分 类 号：R735.7[医药卫生—肿瘤]