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目的在ｉｎｖｉｔｒｏ和ｉｎｖｉｖｏ水平建立致死神经胶质瘤的ＨＳＶ－ｔｋ／ＧＣＶ自杀基因系统，并验证该系统的有效性。方法用携带单纯疱疹病毒胸苷激酶（ＨＳＶ－ｔｋ）基因的重组逆转录病毒，感染大鼠神经胶质瘤细胞Ｃ６，筛选稳定表达ｔｋ的克隆细胞Ｃ６／ｔｋ；经生长抑制试验，比较Ｃ６／ｔｋ细胞对三种核苷类似物ＧＣＶ、ＢＶｄＵ和ＡＣＶ的敏感性；Ｃ６／ｔｋ细胞在裸鼠皮下成瘤，用ＧＣＶ进行治疗并观察疗效。结果ｔｋ基因整合入Ｃ６细胞并在Ｃ６／ｔｋ细胞中稳定表达；生长抑制试验表明，ＧＣＶ是最为有效的原药，Ｃ６／ｔｋ细胞对ＧＣＶ高度敏感，ＩＣ５０＜０．２μｍｏｌ／Ｌ，而野生型Ｃ６细胞和转染空载病毒载体的Ｃ６／０细胞对ＧＣＶ的ＩＣ５０≥１００μｍｏｌ／Ｌ，相差５００多倍。裸鼠ｉｎｖｉｖｏ实验得到相应结果，治疗组肿瘤受到明显抑制和杀伤。结论在ｉｎｖｉｔｒｏ和ｉｎｖｉｖｏ水平，表达ｔｋ基因的肿瘤细胞均被ＧＣＶ有效杀伤，这表明ＨＳＶ－ｔｋ／ＧＣＶ自杀基因系统有可能成为基因治疗脑瘤的有效方法。
Objective To establish a HSV-tk/GCV suicide gene system for lethal gliomas at both invitro and invivo levels and to verify the effectiveness of the system. METHODS: Rat glioma cell C6 was infected with a recombinant retrovirus carrying the herpes simplex virus thymidine kinase (HSV-tk) gene, and the cloned cells stably expressing tk were selected for C6/tk. The growth inhibition test was used to compare C6/ The sensitivity of tk cells to the three nucleoside analogues GCV, BVdU, and ACV; C6/tk cells were subcutaneously tumor-bearing in nude mice, treated with GCV and observed for efficacy. Results tk gene was integrated into C6 cells and stably expressed in C6/tk cells; growth inhibition test showed that GCV is the most effective drug, C6/tk cells are highly sensitive to GCV, IC50 <0.2 μmol/L, and wild type The IC50 of C6 cells and C6/0 cells transfected with no-loaded virus vector to GCV ≥100 μmol/L, a difference of more than 500 times. The corresponding results were obtained in nude mice invivo experiments, and tumors in the treatment group were significantly inhibited and killed. Conclusion In the invitro and in vivo levels, tumor cells expressing the tk gene were effectively killed by GCV, suggesting that the HSV-tk/GCV suicide gene system may be an effective method for gene therapy of brain tumors.