榄香烯复合抗肿瘤疫苗免疫的效应机制.pdfVIP

中国农业科学 2004,37(9):1379-1384 Scientia Agricultura Sinica 榄香烯复合抗肿瘤疫苗免疫的效应机制 金 梅，白 雪, 胡景慧 （辽宁师范大学生命科学院，大连 116029 ） 摘要 ：以小鼠H22腹水型肝癌和L615白血病为实验模型，采用榄香烯或／和热休克等为应激原(stressor)， 对小鼠H22腹水型肝癌细胞和L615白血病细胞进行不同浓度榄香烯、不同时间热休克处理，用透射电镜观察肿瘤 细胞的形态变化；采用直接免疫荧光法流式细胞术(FCM)检测肿瘤细胞凋亡百分率及对肿瘤细胞膜 HSP70、HSP90 表达的影响；进而分离纯化不同应激原(榄香烯、丝裂霉素-C)诱导的H22和L615肿瘤细胞的HSP70－肽复合物， 并用之免疫Balb/c和615小鼠，进行体内抗瘤免疫效应的研究。透射电镜显示，榄香烯或热休克处理都能引起 2 种肿瘤细胞的凋亡和坏死，L615白血病细胞较H22肝癌细胞更敏感，形态学变化更剧烈；流式细胞术检测进一步 证明，榄香烯与热休克都能诱导 2 种肿瘤细胞发生凋亡，两者复合作用更能促进肿瘤细胞的凋亡，在一定范围内 L615细胞凋亡与榄香烯的浓度和热休克的时间呈明显的依赖关系；用榄香烯或MMC 诱导的2种肿瘤细胞分离纯化 的 HSP70－肽复合物给 Balb/c 和 615 小鼠进行主动免疫，免疫组小鼠在 1 个月内全部存活。而 buffer D 免疫对 照组小鼠无一存活。 关键词： 榄香烯； 凋亡； 热休克蛋白 ；抗肿瘤疫苗 Immunity Effects Mechanism Treated with Elemene- Combo Tumor Vaccine JIN Mei ，BAI Xue, HU Jing-hui (Institute of Life Science, Liaoning Normal University, Dalian 116029 ) Abstract: The possible relationship between heat shock protein (HSP) and the molecular mechanisms of active immunization of elemene-combo tumor cell vaccine (TCV) was tudied by using murine H22 ascitic hepatic carcinoma and L615 leukemia as the experimental models. Stressors, such as elemene or/and heat shock, were applied to study the following items: treated with different concentrations of elemene and different time course of heat shock, transmission electron microscopy were used to observe the morphological influence on tumor cells. Direct immunofluorescent flow cytometry (FCM) was used to detect the apoptotic rate of H22 and L615 tumor cells and influence on the expression of membrane HSP70 and HSP90. The membrane HSP70-peptide complex was separated and purified from H22 and