Tca8113细胞exosomes的制备、鉴定及体外抗瘤作用观察

目的:探讨舌癌Tca8113细胞能否分泌exosomes,为口腔肿瘤的免疫治疗寻找新方法。方法:采用超速离心及差异密度梯度离心等方法,从Tca8113细胞中分离并纯化exosomes,透射电镜观察其超微结构,Westernblot分析其表面蛋白成分,最后通过与树突状细胞(dendritic cells,DC)及细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL)等介导的细胞毒性实验来观察exosomes体外抗肿瘤效应。结果:从Tca8113细胞中成功分离并纯化出exosomes,其结构及携带蛋白与其他细胞来源的相似,并发现其分泌的exosomes还携带有肿瘤抗原MAGE蛋白。体外抗瘤实验表明:负载exosomes的DC诱导的CTL对Tca8113细胞的杀伤率明显高于未负载exosomes的DC诱导的CTL及加IL-2培养的T细胞,有显著性差异(P<0.01)。结论:舌癌Tca8113细胞能分泌exosomes,exosomes携带各种与肿瘤免疫相关的蛋白,负载DC后,可以诱导T细胞成为抗原特异性CTL,具有特异杀伤肿瘤细胞的功能,为口腔肿瘤的免疫治疗开拓了新的途径。

Studies on the Isolation, Identification and Anti-tumor Mechanism of Exosomes from Cell Line Tca8113

Objective： To study whether the Tca8113 cell line can secrete exosomes, and identification their anti-tumor potentialities. Methods： Exosomes was isolated and purified from the culture supernatant of Tca8113 cell line by uhraspeed-centrifugation and density gradient centrifugation, and confirmed by transmission electron microscopy and Western blot. Anti-tumor efficacy of the CTL, activated by DC loaded with exosomes, was examined. Results： The CTL induced by DC pulsed with exosomes had significant activity killing Tca8113 cell line compared with CTL induced by DC without exosomes and T cells cultured with IL-2. Conclusions： TcaSll3 cells secrete exosomes. Tumor-derived exosomes may contain tumor special antigen and/or common tumor antigens. The exosome-specific CTL could kill Tca-8113 cells.