单纯疱疹病毒胸苷激酶基因/Ganciclovir系统对脑胶质瘤动物模型的治疗  

HSV_1-tk/GCV GENE THERAPY IN GLIOMA ANIMAL MODEL

作　　者：刘羿男[1] 田竟生[1] 

机构地区：[1]首都医科大学北京神经科学研究所,北京100054

出　　处：《解剖学报》2001年第4期309-313,共5页Acta Anatomica Sinica

基　　金：北京市自然科学基金资助项目 (795 10 0 3 )

摘　　要：目的　探讨单纯疱疹病毒Ⅰ型胸苷激酶基因 (HSV1 tk)结合Ganciclovir(GCV)治疗方案 ,在脑胶质瘤动物模型中基因治疗的疗效以及杀伤肿瘤细胞的机制。　方法　采用Southern杂交法 ,证实逆转录病毒载体生产细胞系pN2 A tk VPC(VPC)带有tk基因并稳定整合到基因组DNA上 ;再将VPC与人脑多形性胶质母细胞瘤细胞系DBTRG 0 5MG(0 5MG)分别以 1∶1、1∶4比例混合接种于BALB c裸鼠皮下 (以只接种 0 5MG细胞作为对照 ) ,建立裸鼠皮下胶质瘤动物模型 ,然后腹腔注射GCV治疗 [30mg (kg·d) ],连续 14d ,治疗后 1周 ,取出瘤块作病理组织学分析。　结果　 1∶1组瘤块较 1∶4组和对照组明显减小 (P <0 0 1) ,且有 30 %的瘤块完全消失 ,提示HSV1 tk GCV对肿瘤细胞有显著杀伤效应。光镜观察可见 ,1∶1组细胞核固缩、溶解甚至破裂等坏死特征 ,说明HSV1 tk GCV在invivo水平杀伤肿瘤细胞是细胞毒杀伤效应。　结论　HSV1 tk GCV系统能有效杀伤肿瘤细胞 。Objective To evaluate the therapeutic effect and the mechanism of herpes simplex virus thymidine kinase Ⅰ type (HSV 1\|tk) and Ganciclovir(GCV) in animal model of human glioma. Methods We demonstrated that tk gene was stably integrated into the genome of retrovirus vector producing cell line pN 2A\|tk/VPC(VPC) by Southern blot hybridization; then we established the nude mice model of glioma by inoculating the VPC and the human glioma cells DBTRG\|05MG(05MG) at 1∶1,1∶4 ratios respectively(only inoculated 05MG cells as control group), then GCV treatment was administrated i.p.,30mg/(kg\5d), twice daily for 14 days. A week later, the nude mice were sacrificed and the pathological sections of tumor tissues were observed by light microscopy. Results The tumor mass of 1∶1 group decreased remarkably compared with 1∶4 group or control group respectively ( P <0 01), and the 30%(6/20) tumor mass were completely regressed, which suggested to have a strong killing effect in vivo. Tumor sections revealed that the nucleus became condensed and then disrupted, which showed necrosis features. Conclusion HSV 1\|tk/GCV can curtail the growth of glioma in nude mice and enhance the killing effect by bystander effect.

关 键 词：脑胶质瘤 单纯疱疹病毒Ⅰ型胸苷激酶 基因治疗 动物模型 Ganciclovir系统 

分 类 号：R739.41[医药卫生—肿瘤] R730.5[医药卫生—临床医学]