结直肠癌腹水来源的exosome提取方法的比较  

The Comparison Between Extraction Methods of Exosomes from Ascites of Colorectal Cancer

作　　者：陈丙辰[1,2] 贺万斌[1,2] 杨旭洋 王蒙[1,2] 杨廷翰[1,2] 伍洋平 魏明天[1,2] 杨金亮[3] 王自强[1,2] 

机构地区：[1]四川大学华西临床医学院,成都610041 [2]四川大学华西医院胃肠外科,成都610041 [3]四川大学生物治疗国家重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2016年第1期102-105,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81172373)资助

摘　　要：目的比较两种从结直肠癌患者腹水中提取纯化exosome的方法,得出最优方法。方法分别采用蔗糖/重水密度梯度超速离心法和Exo-Quick提取法从结直肠癌患者腹水中提取exosome。利用投射电镜检测形态,Western blot技术检测exosome标记蛋白CD63、CD9,使用Nanodrop进行蛋白定量。结果两种方法均可提取exosome,蛋白定量显示Exo-Quick提取法提取exosome蛋白浓度更高,单位体积腹水的提取获得率更高。结论Exo-Quick提取法是一种操作简单、提纯纯度高、提取效率高的提取腹水exosome的方法。Objective To compare the two different methods to isolate the exosome from the ascites of colorectal cancer（CRC）patient and find the efficient one.Methods Exosome from the ascites of CRC patient were isolated by two different methods：density gradient exosome isolation（DG-Exo）and Exo-Quick isolation,and followed by identification with transmission electron microscopy observation and Western blot analysis.And then,Nanodrop was used for protein quantification.Results Exosome were isolated by both of the two methods.The protein concentration of the exosome isolated by the Exo-Quick isolation were higher than that of DG-Exo.Conclusion Exo-Quick isolation can obtain higher purity and more complete exosome from the ascites.

关 键 词：EXOSOME 蔗糖/重水密度梯度超速离心法 Exo-Quick提取法 

分 类 号：R735.34[医药卫生—肿瘤]