双向电泳分析肝癌细胞线粒体的差异表达蛋白  

Differential Expression of Mitochondrial Proteins in Hepatoma Cells Analyzed Using Two-dimensional Electrophoresis

作　　者：李兴[1,2] 潘卫[3] 邱峰[1] 田昆仑[4] 邱宗荫[1] 

机构地区：[1]重庆医科大学医学检验系 [2]贵阳医学院医学检验系,贵州贵阳550004 [3]贵阳医学院医学检验系 [4]第三军医大学野战外科研究所第二研究室,重庆400016

出　　处：《癌症》2005年第11期1327-1331,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金(No.30476021)~~

摘　　要：背景与目的:线粒体在细胞凋亡中扮演关键的角色,对线粒体蛋白质组的研究可能有助于解释癌症的发生、早期诊断以及抗癌药物的研发。本研究对正常肝细胞L02线粒体、表阿霉素干预前后肝癌细胞QGY-7703线粒体进行二维电泳图谱的比较分析,筛选差异表达蛋白斑点。方法:运用蔗糖密度梯度的超离心方法分离纯化线粒体,双向凝胶电泳分离线粒体蛋白质,ImageMaster2D软件对所得图谱进行图像分析。结果:L02细胞线粒体、表阿霉素干预前QGY-7703细胞线粒体、表阿霉素干预后QGY-7703细胞线粒体三种标本超离心后,其纯化线粒体琥珀酸脱氢酶比活力分别提高13.8、11.8、10.2倍。L02细胞线粒体和表阿霉素干预前后QGY-7703细胞线粒体经双向凝胶电泳分辨出206、217、214个蛋白斑点,L02细胞与QGY-7703细胞比较筛选出49个差异表达蛋白斑点;表阿霉素干预前后的QGY-7703细胞进行比较,筛选到29个差异表达蛋白斑点。结论:运用超离心和双向电泳的方法成功筛选出一批在QGY-7703细胞线粒体中差异表达的蛋白斑点。BACKGROUND ＆ OBJECTIVE, Mitochondria play a key role in cell apoptosis. Proteomic analysis of mitochondria will contribute to the discovery of tumorigenic mechanism, early detection of cancer, and findings of anticarcinogens. This study was to identify the differentially expressed mitochondrial proteins in hepatoma cells using two-dimensional gel electrophoresis （2-DE）. METHODS.. Mitochondria were isolated from normal hepatocytes L02, and hepatoma QGY-7703 cells before and after treatment of epirubicin by density gradient centrifugation; the proteins of mitochondrial samples were analyzed with 2-DE. RESULTS. After density gradient centrifugation, the mitochondrial marker enzyme activities were increased by 11.8, 13.8, and 10.2 times, respectively, for L02 cells, untreated QGY-7703 cells, and epirubicin-treated QGY-7703 cells, and the 2-DE profiles identified 206, 217, and 214 protein spots, respectively, from the 3 samples. Compared L02 cells and QGY-7703 cells, 49 differentially expressed protein spots were identified;compared QGY-7703 cells before and after treatment of epirubicin, 29 differentially expressed protein spots were identified. CONCLUSION. A set of differentially expressed protein spots are detected in mitochondria of QGY-7703 cells with density gradient centrifugation and 2-DE.

关 键 词：肝肿瘤 QGY-7703细胞株 L02细胞株 双向电泳 线粒体 差异表达 

分 类 号：R735.7[医药卫生—肿瘤]