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目的 :观察HSV -tk/ACV(HerpesSimplexVirusthymidinekinasegene/Acyclovir)系统基因治疗恶性肿瘤过程中旁观者效应的作用。方法 :HSV -tk基因重组逆转录病毒载体转染PA317细胞为PA317-tk细胞。培养PA317-tk细胞即可获得有感染力的携带HSV -tk基因的病毒颗粒 ,然后转染肝癌细胞株SMMC -772 1细胞为SMMC -tk细胞。SMMC -TK细胞与SMMC - 772 1细胞以不同百分比混合 ,用ACV(10 0ug/ml)培养基培养 ,MTT法检测ACV对各组细胞抑制率 (InhibitRate ,IR) ,根据各组IR与该组SMMC -tk细胞数的百分比值 (IR/SMMC -tk % ,IR/St)的大小判断旁观者效应作用的有无和强弱。结果 :旁观效应试验表明 ,含SMMC -tk细胞 10 % ,2 0 % ,30 % ,4 0 % ,5 0 % ,6 0 % ,70 % ,80 % ,90 % ,10 0 %各组IR/St比值分别为2 .15 ,1.91,1.83,1.4 7,1.30 ,1.11,1.0 2 ,0 .93,0 .84 ,0 .76。 10 %～ 70 %各组IR/St比值均大于 1,存在明显旁观者效应 ,以 10 % ,2 0 % ,30 % ,4 0 %组最强 ,80 %～ 10 0 %三组IR/St比值小于 1,旁观者效应不明显。结论 :研究结果证实用HSV -tk/ACV系统基因治疗恶性肿瘤 ,适宜比例TK+ 细胞时 ,存在明显的旁观者效应 ,TK/细胞为 10 %时旁观者效应最强 ,这对于HSV -TK/ACV系统基因治疗恶性肿瘤具有重要意义。
Objective: To observe the effect of bystander effect of HSV-tk / ACV (HerpesSimplexVirusthymidine kinase gene / Acyclovir) system in the treatment of malignant tumor. Methods: PA317 cells were transfected with PA317-tk cells by HSV-tk gene retroviral vector. The PA317-tk cells were cultured to obtain infectious virus particles carrying HSV-tk gene, and then transfected into hepatoma cell line SMMC -772 1 cells SMMC-tk cells. SMMC-TK cells were mixed with SMMC-772 1 cells in different percentages and cultured in ACV (100ug / ml) medium. Inhibitory rate (IR) of ACV was determined by MTT assay. The percentage of SMMC-tk cells (IR / SMMC -tk%, IR / St) determines the presence and absence of strong and weak bystander effects. Results: Bystander effect test showed that IR / MSCs of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 10% St ratios were 2.15, 1.91, 1.83, 1.4 7, 1.30, 1.11, 1.02, 0.93, 0.84, 0.76, respectively. The IR / St ratios of 10% ~ 70% were higher than 1 in each group, and there was obvious bystander effect. The IR / St ratio of 10%, 20%, 30%, 40% and 80% ~ 100% The ratio is less than 1, bystander effect is not obvious. CONCLUSIONS: The results of the study demonstrate that there is a significant bystander effect when using the HSV-tk / ACV system for the treatment of malignant tumors with a suitable proportion of TK + cells, with the strongest bystander effect at 10% TK / cell, which is most important for HSV-TK / ACV Systemic gene therapy for malignancies is of great importance.