| 应用蛋白质组学和RNAi技术筛选鼻咽癌细胞中p53功能相关蛋白质 |
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| 引用本文： | 孙懿,易 红,杨轶轩,张鹏飞,李茂玉,李建玲,杨 芳,肖志强,陈主初.应用蛋白质组学和RNAi技术筛选鼻咽癌细胞中p53功能相关蛋白质[J].生物化学与生物物理进展,2007,34(7):760-769. |
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| 作者姓名： | 孙懿 易 红 杨轶轩 张鹏飞 李茂玉 李建玲 杨 芳 肖志强 陈主初 |
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| 作者单位： | 1. 中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙,410008;南大学湘雅医学院肿瘤研究所,长沙,410078
2. 中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙,410008 |
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| 基金项目： | 国家重点基础研究发展计划(973计划);教育部跨世纪优秀人才培养计划;国家自然科学基金;湖南省科技厅重大和重点科研项目 |
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| 摘 要： | 为了阐明鼻咽癌中高表达的p53蛋白聚集与失活的机制,高通量地检测与p53功能相关的蛋白质,首先采用RNA干扰(RNAi)技术稳定沉默鼻咽癌细胞系CNE2的p53基因表达,然后用蛋白质组技术研究稳定沉默该基因对鼻咽癌蛋白质表达谱的影响.通过对稳定干扰p53基因后鼻咽癌细胞系CNE2的蛋白质表达谱改变的研究,用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析和电喷雾串联质谱(ESI-Q-TOF-MS)验证鉴定了22个差异表达蛋白质.在这些差异表达蛋白质中,有些是已经报道的p53功能相关蛋白质,如热休克蛋白27(HSP27)、异质性胞核核糖核蛋白K(hnRNPK)、14-3-3σ等,其他可能是新的p53功能相关蛋白质,如eIF4B、TPT1、hnRNPH3、SFRS1等.部分差异表达蛋白质如HSP27、14-3-3σ和GRP75经蛋白质印迹分析技术进行了验证,同时pcDNA3.1-FLAG-p53质粒转染CNE2细胞引起了HSP27、14-3-3σ表达下调,GRP75表达上调.在鼻咽癌细胞中鉴定的22个差异表达蛋白质大致可以分为5类,包括信号传导相关蛋白质、分子伴侣、与转录和翻译相关蛋白质、代谢相关蛋白和细胞结构相关蛋白质,涉及到细胞周期的调控、分子基因表达调控、细胞黏附、细胞代谢等众多事件,它们可能作为p53功能相关蛋白质,为阐明鼻咽癌中p53蛋白聚集及失活的机制提供了重要依据和线索.
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| 关 键 词： | 鼻咽癌 蛋白质组学 RNA干扰 双向电泳 |
| 收稿时间： | 2007/1/16 0:00:00 |
| 修稿时间： | 2007-01-16 |
Identification of Proteins Associated With p53 in Nasopharyngeal Carcinoma Cell Line CNE2 by Proteomics and RNAi Techniques |
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| Sun Yi,YI Hong,YANG Yi-Xuan,ZHANG Peng-Fei,LI Mao-Yu,LI Jian-Ling,Yang Fang,Xiao Zhi-Qiang and Chen Zhu-Chu.Identification of Proteins Associated With p53 in Nasopharyngeal Carcinoma Cell Line CNE2 by Proteomics and RNAi Techniques[J].Progress In Biochemistry and Biophysics,2007,34(7):760-769. |
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| Authors: | Sun Yi YI Hong YANG Yi-Xuan ZHANG Peng-Fei LI Mao-Yu LI Jian-Ling Yang Fang Xiao Zhi-Qiang and Chen Zhu-Chu |
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| Institution: | Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China; Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China; Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China;Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China; Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China |
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| Abstract: | |
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| Keywords: | p53 MALDI-TOF-MS ESI-Q-TOF |
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