论文部分内容阅读
目的 :观察单纯疱疹病毒胸苷激酶基因 (HSV TK)及其底物丙氧鸟苷 (GCV)的基因治疗系统对骨肉瘤细胞株SAOS 2的杀伤作用。方法 :用DOTAP将含有HSV TK基因的真核表达质粒tgCMV HyTK导入骨肉瘤细胞株SAOS 2 ;提取阳性转染细胞 (SAOS 2TK)的总RNA ,用斑点杂交的方法检测HSV TK基因的表达 ;用MTT比色法测定SAOS 2TK细胞对GCV的敏感性及“旁观者效应”。结果 :SAOS 2TK细胞能产生稳定的TK基因的转录产物 ;SAOS 2TK细胞在GCV的作用下呈现明显的生长抑制效应 ,且观察到强效的“旁观者效应”。结论 :HSV TK/GCV系统能够选择性杀死骨瘤细胞
Objective: To observe the killing effect of herpes simplex virus thymidine kinase gene (HSV TK) and its substrate ganosine (GCV) gene therapy system on osteosarcoma cell line SAOS 2. METHODS: The eukaryotic expression plasmid tgCMV HyTK containing HSV TK gene was introduced into osteosarcoma cell line SAOS 2 with DOTAP; total RNA of positive transfected cells (SAOS 2TK) was extracted and the expression of HSV TK gene was detected by dot blot hybridization; MTT assay was used to determine the sensitivity and “bystander effect” of SAOS 2TK cells to GCV. RESULTS: SAOS 2TK cells could produce stable TK gene transcription products; SAOS 2TK cells showed significant growth inhibitory effect under GCV, and a strong “bystander effect” was observed. Conclusion : HSV TK/GCV system can selectively kill osteosarcoma cells