中国病理生理杂志2001，Vol.17Issue(3)：210-214,5.
单纯疱疹病毒胸苷激酶/丙氧鸟苷系统对
人骨肉瘤细胞杀伤作用的研究
The killing effect of herpes simplex virus thymidine kinase/ganciclovior
system on osteosarcoma cells
乔慧 1丘钜世 1张惠忠 2李扬 1郑树森1
作者信息
- 1. 中山医科大学病理教研室,
- 2. 中山医科大学附属肿瘤医院病理科,
- 折叠
摘要
Abstract
AIM: To investigate killing effect of herpes simplex virus thymidine kinase/ganciclovior system(HSV-TK/GCV) on osteosarcoma cell and its mechanisms.METHODS: Recombinant retroviral vector (DORHyTK) containing hygromycin phosphotransferase-thymidine kinase fusion gene(HyTK) was constructed and introduced into human osteosarcoma cell line OS732 with DOTAP. DNA and total RNA extracted from HyTK expressing cells (OS732TK) were tested by PCR and RNA dot blot analysis. A chemosensitivity of OS732TK cells to GCV and “bystander effect” were measured by means of MTT colorimetric assay. Hoeschst 332258 staining and flow cytometric analysis were performed for mechanism of HSV-TK/GCV system gene therapy. RESULTS: DORHyTK was constructed successfully; the HyTK gene existed and expressed in OS732TK cells; All OS732TK cells were killed after 5 days of exposure to 5 mg/L GCV. The “bystander effect” was observed in both a high population and a low population, but the former was stronger than the latter. Hoeschst 332258 staining revealed the characteristic hallmarks of apoptosis, however necrosis also existed. Flow cytometric analysis of DNA content showed a G0-G1 phase blockade. CONCLUSION: HSV-TK/GCV gene therapy system showed its strong killing effects on osteosarcoma cells OS732; The phenomenon of “bystander effect” was also very apparent. GCV exposure induced both necrotic and apoptotic death in HSV-TK expressing cells, and perturbed the cell cycle. HSV-TK/GCV gene therapy system may provide a new therapeutic approach for treatment of osteosarcoma.关键词
骨肿瘤/基因疗法/遗传学分类
医药卫生引用本文复制引用
乔慧,丘钜世,张惠忠,李扬,郑树森..单纯疱疹病毒胸苷激酶/丙氧鸟苷系统对
人骨肉瘤细胞杀伤作用的研究[J].中国病理生理杂志,2001,17(3):210-214,5.
