|
|||||
|
|
| 线粒体DNA缺失A549细胞与其母本细胞核蛋白质组差异分析(英文) | |
| 引用本文： | 赵鹏,张朝晖,仲伟鉴,应贤平,袁准,姚碧云,傅娟玲,周宗灿.线粒体DNA缺失A549细胞与其母本细胞核蛋白质组差异分析(英文)[J].中国药理学与毒理学杂志,2012,26(4):482-488. |
| 作者姓名： | 赵鹏 张朝晖 仲伟鉴 应贤平 袁准 姚碧云 傅娟玲 周宗灿 |
| 作者单位： | 1. 北京大学医学部毒理学系,北京,100191 2. 北京大学医学部毒理学系,北京100191;南华大学环境医学和放射卫生院,湖南衡阳421001 3. 上海市疾病预防控制中心毒理科,上海,200336 |
| 基金项目： | The,project,supported,by,National,Natural,Science,Foundation,of,China,National,Natural,Science,Foundation,of,China,Fundamental,Research,Funds,for,Central,Universities,国家自然科学基金,国家自然科学基金,北京大学医学部新教师科研启动基金 |
| 摘 要： | 目的比较线粒体DNA(mtDNA)缺失A549细胞(Rho0细胞)与其母本细胞(Rho+细胞)核蛋白表达谱,并探讨细胞核对线粒体功能缺陷的应答反应。方法二维凝胶电泳(2-DE)和表面增强激光法解吸电离-飞行时间(SELDI-TOF)蛋白芯片测定Rho0细胞和Rho+细胞核蛋白表达谱,基质辅助激光解吸电离-飞行时间(MALDI-TOF)质谱结合数据库检索鉴定差异表达的蛋白点,Western印迹法测定核磷蛋白和P53表达,激光共聚焦显微镜测定线粒体膜电位。结果 2-DE显示Rho0细胞核中11个蛋白点表达下调,21个蛋白点表达上调。基于NP20蛋白质芯片的SELDI-TOF质谱分析发现4个蛋白质峰在Rho0细胞核中明显下降。其中1个表达下调的蛋白点被鉴定为eIF-6,4个表达上调的蛋白点被鉴定为核磷蛋白,SFRS1,SFRS3和hnRNP G。Western印迹实验结果显示,Rho0细胞中核磷蛋白表达增加。P53和线粒体膜电位(MMP)测定结果显示,Rho0细胞中P53表达高于Rho+细胞,两种细胞MMP基本一致。结论 mtDNA缺失诱导了细胞核蛋白质组改变。Rho0细胞可以作为研究线粒体与核交互作用的模型。 |
| 关 键 词： | 核蛋白质组 基因 p53 膜电位 线粒体 |
| 收稿时间： | 2011-6-1 |
| 修稿时间： | 2012-3-19 |
Comparative analysis of nuclear proteomes in mitochondrial DNA-depleted A549 cells and their parental cells |
|
| ZHAO Peng , ZHANG Zhao-hui , ZHONG Wei-jian , YING Xian-ping , YUANN Zhun , YAO Bi-yun , FU Juan-ling , ZHOU Zong-can.Comparative analysis of nuclear proteomes in mitochondrial DNA-depleted A549 cells and their parental cells[J].Chinese Journal of Pharmacology and Toxicology,2012,26(4):482-488. | |
| Authors: | ZHAO Peng ZHANG Zhao-hui ZHONG Wei-jian YING Xian-ping YUANN Zhun YAO Bi-yun FU Juan-ling ZHOU Zong-can |
| Institution: | 1. Department of Toxicology, Health Science Center, Peking University, Beijing 100191, China;2. Institute of Environmental Medicine and Radiation Hygiene, University of South China, Hengyang 421001, China;3. Department of Toxicology, Shanghai Municipal Center for Disease Control & Prevention, Shanghai 200336, China |
| Abstract: | OBJECTIVE To investigate the nuclear proteomes in mitochondrial DNA (mtDNA)-depleted A549 cells (Rho0 cells) and their parental cells (Rho+ cells), and to learn more about the nuclear responses to mitochondrial dysfunction. METHODS The nuclear proteomes of Rho0 and Rho+ cells were characterized by two dimensional electrophoresis (2-DE) and SELDI-TOF ProteinChip technologies, the differentially expressed protein-spots were identified by MALDI-TOF mass spectrum (MS), the nucleophosmin and P53 expression were detected by Western blotting assay, and the mitochondrial membrane potential (MMP) was measured by the laser scanning confocal microscope. RESULTS 2-DE results showed 11 protein-spots were down-regulated and 21 protein-spots were up-regulated in Rho0 cell nuclei. SELDI-TOF MS analysis with NP20 ProteinChips revealed 4 protein-peaks decreased in Rho0 cell nuclei. One down-regulated protein-spot was identified as eIF-6, and 4 up-regulated protein-spots were identified as nucleophosmin, SFRS1, SFRS3 and hnRNP G, respectively. The increased expression of nucleophosmin in Rho0 cells was verified by Western blotting. Based on the clues from proteomic analysis, P53 expression in Rho0 cells was higher than in Rho+ cells, and MMP was consistent in Rho+ and Rho0 cells. CONCLUSIONmtDNA-depletion induces nuclear proteome alteration. Rho0 cells can be used as a model to study the cross-talk between mitochondrion and nucleus. |
| Keywords: | nuclear proteomics genes p53 membrane potential mitochondrial |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |
| 点击此处可从《中国药理学与毒理学杂志》浏览原始摘要信息 | |
| 点击此处可从《中国药理学与毒理学杂志》下载免费的PDF全文 | |