胶质瘤细胞来源exosome的鉴定及其蛋白组成研究  

Identification of glioma derived exosome and its proteomic analysis

作　　者：杜嘉瑞[1] 张路遥[1] 杨洪发[1] 别黎[1] 徐惠勇[2] 陈立兵[2] 鞠砚[1] 

机构地区：[1]吉林大学第一附属医院神经外科,吉林长春130021 [2]吉林省九台市中医院

出　　处：《中国实验诊断学》2008年第1期72-76,共5页Chinese Journal of Laboratory Diagnosis

摘　　要：目的证实胶质瘤细胞是否可分泌exosome,并初步分析其蛋白组成,探讨胶质瘤来源exosome的潜在免疫调节功能,从而为进一步利用exosome对胶质瘤行免疫治疗提供理论依据。方法差速离心法从胶质瘤细胞培养上清液中提纯exosome,用透射电镜鉴定,利用SDS-PAGE初步分析exosome蛋白组成,并用western-blot验证hsp70的存在。结果胶质瘤细胞可以产生大小均一的囊泡性结构,即exosome,其平均直径约100 nm。exosome内所含蛋白质经初步分析,其表观分子量介于270-30 kD之间。经western-blot证实其内含有hsp70分子。结论胶质瘤细胞也可分泌exosome,其一般特性与已报道的exosome一致,蛋白组成与其他细胞来源的exosome具有共性,且含有hsp70分子,具有一定的免疫调节功能,可以为抗胶质瘤免疫治疗提供理论基础。Objective Our aim was to prove that ghoma can also secrete exosome-like vesicles,and initially analyze the protein composition in order to offer a theoretical fotmdation for its utilization in anti-glioma immunotherapy. Methods Exosome was purified by differential altracentrifugation, and the electron microscopy was used to check the purity of exosomes. Proteins were separated by one-dimensional SDS-PAGE, and protein bands were analyzed by software. And hsp70 was identified by western-blot. Results Gliorna cell could also secrete exosome-like vesicles. The morphologic characterization of exosome was： spherical or oval-shaped membrane vesicles, syngeneous in size with an average diameter about 100 nm. The data obtained from SDS-PAGE and software showed that the appearance molecular weights （MW） of all the proteins contained by exosomes were between 270 kD and 30kD, which is consistent with previous studies. Hsp70 was detected to present in glioma derived exosome by western-blotting. Conclusion It was confirmed that ghoma cells also bad the ability to secrete exosome-like vesicles. All the data obtained from this experiment and other articles that have reported revealed a common constituent between exosomes from glioma cells and other cells. The presence of hsp70 reveals some properties of immunomodulation for glioma derived exosome.

关 键 词：EXOSOME 胶质瘤 HSP70 透射电镜 WESTERN-BLOT 

分 类 号：R739.4[医药卫生—肿瘤]