CAJ | 학술논문

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。
목적탐토선태병독Tianjin주결손간우과립（ defective interfering particles ，DI과립）체내외유도대서뇌효질류세포C6조망적작용。방법장불동적도선태병독Tianjin주DI과립분별여대서뇌효질류세포C6작용불동시간，이배양기작위음성대조、완정병독작위양성대조，통과DNA편단경지당응효전영、TUNEL염색、AnnexinⅤ-FITC/PI표기류식세포의분석등검측세포조망정황。건립대서피하효질류모형，통과측량종류대소관찰DI과립억류작용，병리절편HE염색관찰종류조직병리변화，TUNEL법검측종류조직세포조망정황。결과 C6세포재체외경DI과립유도후， DNA편단경지당응효전영정계제상；류식세포의급TUNEL검측현시，DI과립조여완정병독조세포조망솔명현증고，차정시간-제량의뢰형。동물실험결과현시，DI과립화완정병독균가명현억제종류생장；종류조직병리절편HE염색현시DI과립조화완정병독조류결절내류세포교소；TUNEL원위세포조망검측현시DI과립조화완정병독조조망세포명현증가，이상결과여음성대조조비교차이균유통계학의의（P＜0．01）。결론선태병독Tianjin주DI과립재체내외균능인기대서뇌효질류C6세포조망，차정시간-제량의뢰형，제시DI과립유보조치료뇌효질류적가능성。
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 (P<0.01) as indicated by flow cytometry assay and TUNEL assay .In vivo, DI par-ticles could markedly inhibit the growth of the tumors in comparison with saline control group .There were fe-wer tumor cells in tumor nodules in DI particles group or intact virus group as shown by histological examina -tion.The TUNEL assay showed that the apoptosis rate of tumor tissues injected with DI particles or intact vi -rus was much higher than that of the saline group (P<0.01), but there was no significant difference between the DI particles group and the intact virus group (P>0.05).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.