仙台病毒 Tianjin 株缺陷干扰颗粒体内外诱导大鼠脑胶质瘤细胞凋亡的研究  

Study on the apoptosis of rat glioma cells induced by defective interfering particles of Sendai virus strain Tianjin

作　　者：韩哲[1] 周洪经[1] 赵杰[1] 贾浩波[1] 陈曦腾[1] 石立莹[1] 

机构地区：[1]天津医科大学基础医学院微生物学教研室,300070

出　　处：《中华微生物学和免疫学杂志》2013年第9期677-682,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目

摘　　要：目的探讨仙台病毒Tianjin株缺损干扰颗粒（ defective interfering particles ，DI颗粒）体内外诱导大鼠脑胶质瘤细胞C6凋亡的作用。方法将不同滴度仙台病毒Tianjin株DI颗粒分别与大鼠脑胶质瘤细胞C6作用不同时间，以培养基作为阴性对照、完整病毒作为阳性对照，通过DNA片段琼脂糖凝胶电泳、TUNEL染色、AnnexinⅤ-FITC/PI标记流式细胞仪分析等检测细胞凋亡情况。建立大鼠皮下胶质瘤模型，通过测量肿瘤大小观察DI颗粒抑瘤作用，病理切片HE染色观察肿瘤组织病理变化，TUNEL法检测肿瘤组织细胞凋亡情况。结果 C6细胞在体外经DI颗粒诱导后， DNA片段琼脂糖凝胶电泳呈阶梯状；流式细胞仪及TUNEL检测显示，DI颗粒组与完整病毒组细胞凋亡率明显增高，且呈时间-剂量依赖型。动物实验结果显示，DI颗粒和完整病毒均可明显抑制肿瘤生长；肿瘤组织病理切片HE染色显示DI颗粒组和完整病毒组瘤结节内瘤细胞较少；TUNEL原位细胞凋亡检测显示DI颗粒组和完整病毒组凋亡细胞明显增加，以上结果与阴性对照组比较差异均有统计学意义（P＜0．01）。结论仙台病毒Tianjin株DI颗粒在体内外均能引起大鼠脑胶质瘤C6细胞凋亡，且呈时间-剂量依赖型，提示DI颗粒有辅助治疗脑胶质瘤的可能性。Objective To investigate the apoptosis of rat glioma C 6 cells induced by defective in-terfering（ DI） particles of Sendai virus strain Tianjin .Methods Rat glioma C6 cells were treated with dif-ferent titers of DI particles of Sendai virus strain Tianjin in vitro with culture media as negative control and intact virus as positive control .At different time point , cells were collected and their apoptosis was detected by DNA gel electrophorsis , TUNEL assay and AnnexinⅤ/PI double-labeled flow cytometry .The C6 glioma-bearing rat model was established and then treated with three intratumoral injections of DI particles , intact virus or saline three times at interval of two days .The antitumor effects of ID particles were evaluated through daily measuring of the tumor size .Hematoxylin-eosin（ HE） staining was used to observe the patho-logical changes in tumor tissues .TUNEL assay was performed to detect the apoptosis of tumor tissues .Re-sults Rat glioma C6 cells treated with DI particles or intact virus in vitro showed typical DNA ladder pattern in agarose gel electrophoresis in a time-and dose-dependent manner .With the intervention of DI particles , the apoptosis rate of C6 cells showed a time-and dose-dependent manner and was significantly higher than that of the control group （P0.05）.Conclusion The DI particles of Sendai virus strain Tianjin could induce apoptosis of rat glioma C 6 cells in a time-and dose-dependent manner both in vitro and in vivo, suggesting that the DI particles might be applicable for the treatment of neurogliocytoma in the future.

关 键 词：仙台病毒Tianjin株 缺损干扰颗粒 大鼠脑胶质瘤细胞C6 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]