期刊论文

Establishment of the conditions for the extraction of nuclear proteins from the primary focus of nasopharyngeal carcinoma

中文

医学研究生学报

1008-8199

2010

23

4

347-351

10.16571/j.cnki.1008-8199.2010.04.024

30672738,30572408:国家自然科学基金 06WK3016:湖南省科技厅国际科技合作项目 2007TP4021:湖南省科技厅项目 08C637:湖南省教育厅一般项目

本校为第一机构

目的鼻咽癌细胞蛋白组分复杂,分离并分析其细胞核蛋白有助于进行肿瘤细胞蛋白质组学研究。文中建立并优化从原发鼻咽癌活检组织标本中提取肿瘤细胞核蛋白的方法。方法采用差速离心结合蔗糖密度梯度离心方法分离和富集鼻咽癌患者活检组织中肿瘤细胞的细胞核,用甲醇/三氯甲烷法和丙酮沉淀法提取核蛋白,用Western blot方法鉴定细胞核蛋白。结果分离细胞核和提取核蛋白至少需要200mg鼻咽癌组织。用差速离心和蔗糖密度梯度离心方法进行细胞核抽提,用甲醇/三氯甲烷和丙酮沉淀法提取的细胞核蛋白具有较高纯度,基本无其他细胞器污染,可用于鼻咽癌原发病灶中细胞核蛋白的提取分离。结论建立了从鼻咽癌原发...

Objective The total protein components of nasopharyngeal carcinoma cells is very complex, and an insight into the isolated nuclear proteins may be helpful for the study on wholecell proteomics. This article aimed to explore the establishment of the conditions for the extraction of nuclear proteins from the primary focus of nasopharyngeal carcinoma （NPC）. Methods The nuclei of nasopharyngeal tissues from the primary focus of NPC were extracted by differential centrifugation and sucrose density-gradient centrifugation. Methanol/trichloromethane...