顺铂联合exosomes抗小鼠肝癌效应的实验研究  

Experimental studies on anti-mouse hepatocellular carcinoma effects of cisplatin combined with exosomes

作　　者：汪少华[1] 沈宜[1] 李静[1] 向自武[1] 范维珂[1] 陈黎[1] 

机构地区：[1]重庆医科大学基础医学院病理生理教研室干细胞与组织工程研究室,重庆400016

出　　处：《细胞与分子免疫学杂志》2009年第1期49-52,共4页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:评价顺铂(DDP)与exosomes联用的抗肿瘤效果,研究其可能机制。方法:采用MTT法检测顺铂对小鼠肝癌H22细胞增殖的影响,用H22源exosomes瘤苗免疫小鼠,3H-TdR释放法检测exosomes诱导产生的CTL活性及顺铂对CTL杀伤的增敏作用,RT-PCR检测顺铂作用后H22细胞中Fas及exosomes免疫后脾淋巴细胞FasL的mRNA表达水平,Western blot检测顺铂对H22细胞Fas的蛋白表达水平的影响。以小鼠肝癌H22细胞接种BALB/c小鼠建立动物模型,观察顺铂联合exosomes治疗对小鼠生存期的影响。结果:顺铂抑制H22细胞生长呈量效关系;exosomes免疫小鼠可诱导产生针对H22细胞的CTL反应,经2.5mg/L顺铂预处理24h的H22细胞对CTL杀伤的敏感性增强(P<0.05)。顺铂在mRNA和蛋白水平,显著增强H22Fas的表达;exosomes免疫小鼠后,脾淋巴细胞FasL表达增加。顺铂联合exosomes组小鼠生存期较单独治疗组及对照组明显延长(P<0.05)。结论:顺铂与exosomes联合治疗有协同抑制肿瘤的作用,产生协同作用的机制与增强CTL活性有关。AIM： To study the anti-tumor effects and mechanisms of cisplatin （DDP） combined with exosomes. METHODS： The cell-growth inhibition of DDP on H22 cells was analyzed by MTT assay. The mice were immunized by H22 cell-derived exosomes. The exosomes elicited CTL-specific cytotoxicity and DDP enhanced cytotoxicity in combination with CTL were detected by ^3 H-TdR release assay. The effect of DDP on mRNA and protein levels of Fas in H22 was analyzed using RT-PCR and Western blot. The expression of FasL on spleen lymphocyte was determined by RT-PCR. BALB/c mice inoculated with hepatoma carcinoma cell line H22 were used as tumor models. The mice received exosomes solely or in combination with DDP. The survival of the mice was observed. RESULTS： DDP inhibited H22 cell in a dose-dependant manner. The exosomes elicited CTL-specific cytotoxicity was enhanced by DDP （ P 〈 0.05）. RT-PCR and Western blot showed Fas increased gradually after administering DDP on H22 cells. RT-PCR also indicated the mRNA level of FasL on mice spleen lymphocyte increased after immunized by exosomes. Compared with other groups, the combination group（ DDP plus exosomes）could statistically prolong the survival time（ P〈 0.05）. CONCLUSION： The therapy of DDP combined with exosomes had significant synergistic effect against tumor. The mechanism of synergistic effect includes enhancement of CTL activity.

关 键 词：顺铂 EXOSOMES 联合治疗 肿瘤疫苗 

分 类 号：R730.5[医药卫生—肿瘤] R392.1[医药卫生—临床医学]