鼻咽癌细胞株6-10B质膜的分离及其蛋白质表达谱分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (8): 176-183.

鼻咽癌细胞株6-10B质膜的分离及其蛋白质表达谱分析

张鹏飞¹, 曾谷清^1，3, 汤参娥¹, 易红¹, 梁宋平², 陈主初¹, 肖志强¹

（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；2.湖南师范大学生命科学学院，长沙 410081；3.南华大学医学院外总教研室，衡阳 421001）

收稿日期:2012-12-20 修回日期:2013-08-11 出版日期:2013-08-11 发布日期:2013-09-02
作者简介:张鹏飞，男，博士，副教授，研究方向：鼻咽癌等恶性肿瘤的蛋白质组学；E-mail：zhangpf690421@csu.edu.cn
基金资助:
国家重大新药创制资助项目（2012ZX09303013），国家自然科学基金资助项目（81272609），湖南省卫生厅科研基金资助项目（B2010-004）

Separation of Plasma Membrane from Nasopharyngeal Carcinoma Cell Line 6-10B and Preliminary Analysis of Its Protein Profile

Zhang Pengfei¹, Zeng Guqing^1，3, Tang Can’e¹, Yi Hong¹, Liang Songping², Chen Zhuchu¹, Xiao Zhiqiang¹

（1. Key Laboratory of Cancer Proteomics of Chinese Ministry of Health，Xiangya Hospital，Central South University，Changsha 410008；2. College of Life Sciences，Hunan Normal University，Changsha 410081；3. Department of General Surgery and Operative Surgery，School of Medicine, University of South China, Hengyang 421001）

Received:2012-12-20 Revised:2013-08-11 Published:2013-08-11 Online:2013-09-02

摘要/Abstract

摘要： 建立鼻咽癌（Nasopharyngeal carcinoma，NPC）细胞株6-10B的质膜蛋白质表达谱，为鼻咽癌癌变机制研究提供有益数据资料。运用差速离心联合双水相的方法纯化NPC细胞6-10B质膜，Western blot 印迹和电镜检测验证质膜纯度，一维凝胶电泳分离质膜蛋白质，液相色谱/串联质谱结合生物信息学鉴定质膜蛋白质，肿瘤差异蛋白质组数据库（dbDEPD）搜索质膜蛋白质与其他肿瘤的相关性。共鉴定到406种非冗余蛋白质，质膜蛋白质和质膜相关蛋白共255个（占62.8%），dbDEPD搜索显示145种质膜蛋白质中的46个与其他肿瘤相关，其中18个质膜蛋白质与其他肿瘤转移相关，进一步分析发现质膜蛋白质CD225、CD104 和p63与同属头颈部肿瘤的口腔癌转移相关，提示它们也可能是NPC潜在的转移相关蛋白。本研究建立了NPC细胞株6-10B的质膜蛋白质表达谱数据库，为NPC癌变分子机制研究提供有价值的数据和信息。

关键词: 鼻咽癌, 6-10B, 质膜, 蛋白质组, 双水相, 质谱

Abstract: To establish plasma membrane（PM）protein profile of 6-10B nasopharyngeal carcinoma（NPC）cell, and accumulate valuable data for understanding the carcinogenesis mechanism of NPC. First of all, two-phase partition in combination with differential velocity centrifugation was used to purify PM of 6-10B cell. Then, purified PM were detected the purity by Western blot and electron microscopy. Next, SDS-PAGE was used to isolate PM proteins. After that, PM proteins were identified using mass spectrometry and bioinformatics. Finally, identified PM proteins were determined relevance with other tumors in human cancers database（dbDEPD）searching. A total of 406 proteins of 6-10B cell were identified, and 255 proteins were PM and membrane-associated proteins（accounting for 62.8%）. 46 kinds of proteins in 145 identified PM proteins were tumor-related proteins, of which 18 proteins associated with tumor metastasis. Further analysis found that CD225, CD104 and p63 were oral cancer（belong to head and neck cancer）metastasis related proteins, suggesting that they may be potential metastasis associated proteins in NPC. A PM protein profile of NPC 6-10B cell has been established, which provides useful information to investigate carcinogenesis mechanism of NPC.

Key words: Nasopharyngeal carcinoma, 6-10B cell, Plasma membrane, Proteomics, Aqueous two-phase partition, Mass spectrometry

张鹏飞, 曾谷清, 汤参娥, 易红, 梁宋平, 陈主初, 肖志强. 鼻咽癌细胞株6-10B质膜的分离及其蛋白质表达谱分析[J]. 生物技术通报, 2013, 0(8): 176-183.

Zhang Pengfei, Zeng Guqing^，, Tang Can’e, Yi Hong, Liang Songping, Chen Zhuchu, Xiao Zhiqiang. Separation of Plasma Membrane from Nasopharyngeal Carcinoma Cell Line 6-10B and Preliminary Analysis of Its Protein Profile[J]. Biotechnology Bulletin, 2013, 0(8): 176-183.

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