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目的 体外观察腺病毒载体介导的单纯疱疹病毒胸苷激酶基因及羟基无环鸟苷系统 (ADV -HSV -Tk/GCV)对人腺样囊性癌细胞 (ACC -2及ACC -M )的杀伤作用。方法 分别将人腺样囊性癌细胞株 (ACC -2及ACC -M )用腺病毒构建的绿色荧光蛋白按 1∶10 ,1∶2 0 ,1∶3 0转染 ,在 12～ 3 6h内用荧光显微镜观察细胞的转染率。MTT法观察ADV -HSV -TK在不同感染复数 (5～ 60 )以及不同浓度GCV处理时对ACC -2及ACC -M的细胞杀伤作用以及“旁观者效应”。结果 体外研究发现 ,ACC -2及ACC -M均能被腺病毒 -绿色荧光蛋白转染。最小感染复数为 10～ 2 0 ,转染率与感染复数呈正相关。在感染复数为 5 ,GCV浓度为 2 5 μg/ml时 ,能够发挥细胞杀伤效应且出现剂量依赖性。MTT法检测发现经ADV -HSV -TK处理后 ,当GCV浓度为 2 5 μg/ml或高于该浓度时ACC -2及ACC -M的细胞抑制率为 40 % -60 % ,而当GCV单独作用时则无此作用。转染ADV -HSV -TK的细胞与未转染细胞按不同比例混合提示“旁观者效应”的存在 ,且转染细胞的比例越高 ,被杀伤的细胞比例越大。结论 腺病毒载体介导的单纯疱疹病毒胸苷激酶基因及羟基无环鸟苷系统 (ADV -HSV -TK/GCV)在体外能够有效抑制腺样囊性癌细胞 ,然而 ,其细胞杀伤效应及“旁观者效应”仍有待提高
Objective To observe the effect of adenovirus vector-mediated herpes simplex virus thymidine kinase gene and ADV-HSV-TK / GCV on human adenoid cystic carcinoma cells (ACC-2 and ACC-M) Killing effect. Methods Human adenoid cystic carcinoma cell lines (ACC-2 and ACC-M) were transfected with adenovirus-containing green fluorescent protein (1:10, 1:20, 1:30, respectively) Fluorescence microscopy was used to observe the transfection efficiency of cells. MTT assay was used to observe the cytotoxicity of ADV-HSV-TK against ACC-2 and ACC-M cells and the “bystander effect” at different concentrations (5 to 60) and different concentrations of GCV. Results In vitro studies showed that both ACC-2 and ACC-M were transfected with adenovirus-GFP. The minimum number of infection was 10 ~ 20, and the infection rate was positively correlated with the number of infection. At a multiplicity of infection of 5 and a GCV concentration of 25 μg / ml, the cell killing effect was demonstrated in a dose-dependent manner. MTT assay showed that after ADV-HSV-TK treatment, the cell inhibition rates of ACC-2 and ACC-M were 40% -60% when the GCV concentration was 25 μg / ml or higher, while when GCV alone When the role of no such role. Cells transfected with ADV-HSV-TK were mixed with non-transfected cells at different ratios to suggest the presence of “bystander effect”, and the higher the proportion of transfected cells, the greater the proportion of cells killed. Conclusions Adenoviral vector-mediated herpes simplex virus thymidine kinase gene and hydroxyl acyclovir system (ADV-HSV-TK / GCV) can effectively inhibit adenoid cystic carcinoma cells in vitro, however, its cytotoxicity and “ Bystander effect ”still needs to be improved