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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 ,并可能通过旁观者效应扩大肿瘤杀伤效应
Objective To investigate the efficacy of gene therapy for herpes simplex virus type 1 thymidine kinase gene (HSV1 tk) in combination with Ganciclovir (GCV) therapy in animal models of glioma and the mechanism of killing tumor cells. Methods The Southern hybridization method was used to confirm that the retroviral vector-producing cell line pN2 A tk VPC (VPC) had a tk gene and was stably integrated into the genomic DNA; then the VPC and human brain glioblastoma multiforme cell line DBTRG 0 MG (0 5 MG) was inoculated subcutaneously into BALB c nude mice at a ratio of 1:1 and 1:4 (using only 0 5 MG cells as a control) to establish an animal model of subcutaneous glioma in nude mice, which was then injected intraperitoneally with GCV. [30 mg (kg·d)] for 14 consecutive days. One week after treatment, the tumor mass was removed for histopathological analysis. Results Compared with 1:4 group and control group, tumor mass in 1:1 group was significantly decreased (P < 0.01), and 30% of tumor mass disappeared completely, suggesting that HSV1 tk GCV has a significant killing effect on tumor cells. Observed by light microscope, the 1:1 group of necrotic features such as cell nucleus pyknosis, dissolution or even rupture, indicating that cytotoxic effects of HSV1 tk GCV in killing tumor cells at in vivo level. Conclusion The HSV1 tk GCV system can effectively kill tumor cells and may increase tumor killing effect by bystander effect.