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目的探讨恶性胶质瘤自杀基因治疗中旁效应的机理。方法野生型C6鼠胶质瘤细胞与基因修饰的C6tk细胞不同比例混合，在GCV作用下体外共培养；C6与C6tk细胞1：1混合大鼠颅内接种，观察肿瘤治疗反应；收集发生凋亡的C6tk细胞的条件培养基．经不同孔径的滤膜过滤，滤液及0220μm滤膜上的物质重新加入野生型瘤细胞，观察细胞存活，送检电镜。结果C6细胞的抑制率随C6tk细胞混入比例的增高而增高并具有GCV剂量依赖性（P＜0.01）；实验组比对照组瘤龄二周时肿瘤体积明显减小（P＜0.01），存活期延长；和溶性物质无细胞毒作用，0.22μm滤膜上物质可使野生型瘤细胞存活率下降（P＜0.01）。电镜观察到瘤细胞对毒性囊泡的吞噬，并经历凋亡。结论野生型瘤细胞对基因修饰细胞凋亡囊泡的吞洚是旁效应的原因，凋亡囊泡的内容最大可能是毒性核酸。
Objective To investigate the side effect mechanism of suicide gene therapy in malignant gliomas. Methods Wild-type C6 murine glioma cells were mixed with genetically modified C6tk cells in different ratios and co-cultured under GCV in vitro. C6 and C6tk cells were mixed in a 1:1 manner and intracranially inoculated to observe tumor treatment response; apoptosis was collected. Conditioning medium for C6tk cells. After filtration through different pore size filters, the filtrate and the substance on the 0220 μm filter were reintroduced into the wild-type tumor cells to observe cell survival and examined by electron microscopy. Results The inhibitory rate of C6 cells increased with the increase of the proportion of C6tk cells and was GCV dose-dependent (P<0.01). The tumor volume of the experimental group was significantly reduced (P<0.01) compared with the control group at 2 weeks of tumor age (P<0.01). Prolonged; and soluble substances without cytotoxicity, 0.22μm filter material can reduce the survival rate of wild-type tumor cells (P <0.01). Electron microscopy observed that tumor cells phagocytosed toxic vesicles and experienced apoptosis. Conclusion The wild-type tumor cells are responsible for the side effect of the apoptotic vesicles of genetically modified cells. The content of apoptotic vesicles may be toxic nucleic acid.