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中文摘要：
目的分离纯化膀胱移行细胞癌T24细胞来源的exosome,对其形态学和蛋白组成进行初步分析,为进一步利用exosome治疗膀胱移行细胞癌奠定基础。方法采用超滤和蔗糖密度梯度离心法分离T24细胞释放的exosome,电镜观察形态大小,SDS-PAGE分离蛋白并用软件分析,Western blot检测热休克蛋白70（HSP70）、细胞黏附分子（ICAM-1）、细胞角蛋白20（CK20）的表达。结果成功的分离出了exosome,其大小为30～90nm,类圆碟形小囊泡,Western blot证实exosome表达HSP70、ICAM-1和CK20分子。结论 T24细胞可以分泌exosome,并负载了HSP70、ICAM-1等免疫相关蛋白,为肿瘤免疫治疗提供了理论基础。
英文摘要：
Objective To separate and purify exosome derived from transitional cell carcinoma of bladder of T24 cell,and initially analyze the morphology and protein composition in order to offer a foundation for its utilization in immunotherapy.Methods The exosomes secreted by T24 cell were purified by ultracentrifugation and sucrose gradient centrifugation,and the electron microscope was used to check the exosome.Proteins were separated by SDS-PAGE,and protein bands were analyzed by software. HSP70,ICAM-1 and CK20 were identified by western blot.Results Exosomes were successfully purified by ultracentrifugation and sucrose gradient centrifugation. The exosomes were syngeneous in size about 30~90nm, and were saucer-shaped membrane vesicles. HSP70,ICAM-1 and CK20 were detected by western blot.Conclusion It was confirmed that T24 cell can secrete exosome,and the presence of immune proteins of HSP70 and ICAM-1 offer a theoretical foundation for anti-tumor in immunotherapy.
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