有丝分裂纺锤体及中间小体提取方法的研究  

Study on mitotic spindle and midbody extraction

作　　者：武岩[1] 潘丽娜[2] 朱长军[2] 姜伟[1] 詹启敏[1] 童彤[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤医院肿瘤研究所、分子肿瘤学国家重点实验室,北京100021 [2]天津师范大学细胞与分子系统生物学重点实验室,300387

出　　处：《国际生物医学工程杂志》2014年第2期76-80,I0001,共6页International Journal of Biomedical Engineering

基　　金：国家重大科学研究计划（2010CB910703）;国家自然科学基金面上项目（31271485）;2011年教育部“新世纪优秀人才支持计划”（NCET-11-1066）

摘　　要：目的 有丝分裂纺锤体和中间小体均是以在细胞有丝分裂进程中出现的微管为结构基础的暂时性结构,对细胞有丝分裂的顺利进行起着至关重要的作用.本研究旨在建立成熟高效的提取细胞内有丝分裂纺锤体和中间小体的方法.方法 通过细胞周期同步化方法使细胞内出现有丝分裂纺锤体或中间小体,运用低渗肿胀和甘油梯度离心的原理提取纺锤体和中间小体.结果 经Western Blot和细胞免疫荧光染色法鉴定,证实提取物中含有有丝分裂纺锤体和中间小体.结论 建立从人宫颈癌细胞Hela中提取有丝分裂纺锤体及中间小体的方法,可为鉴定有丝分裂纺锤体及中间小体上的蛋白分子垫定实验基础,同时也对研究肿瘤细胞有丝分裂的分子调控机制具有重要意义.Objective Mitotic spindle and midbody are both microtubule-based temporary structures during cell growth and play essential roles in mitosis. The purpose of this study was to establish a mature and efficient method to extract mitotic spindle and midbody. Methods Through the cell cycle synchronization method, mitotic spindle or midbody was made appear inside cells. Low permeability swelling and glycerol gradient centrifugation principles were then used to extract spindle and midbody. Results By Western Blot and immunofluorescence staining, the extracts were identified as mitotic spindle and midbody. Conclusions The successful extraction of mitotic spindle and midbody from synchronized Hela cells will provide foundation for identifying the proteins located in cell during mitosis, and be of great significance to the study of molecular regulation mechanisms of mitosis and tumorigenesis.

关 键 词：有丝分裂纺锤体 中间小体 细胞同步化 有丝分裂 

分 类 号：R730.2[医药卫生—肿瘤]