HSV-tk/GCV系统杀伤ACC-M细胞的机理研究

THE MECHANISM OF HUMAN ADENOID CYSTIC CARCINOMA CELL STRAIN (ACC M) CELL DEATH INDUCED BY HSV tk/GCV GENE THERAPY SYSTEM

目的 为探索HSV-tk/GCV(单纯疱疹病毒胸腺嘧啶核苷激酶基因/羟甲基无环鸟苷)基因治疗系统杀伤人高转移性腺样囊性癌细胞ACC-M 的机理。方法 采用荧光染色荧光显微镜观察、细胞DNA 电泳、流式细胞仪检测等方法。结果 研究发现,荧光染色有明显的细胞凋亡小体和染色质边聚现象;在GCV浓度较大(400 μm ol/L)作用时间较长(4 天)时有较明显的DNA 梯状现象;而FCM 发现在400 μm ol/LGCV 作用下有极明显的亚G1 峰。结论 HSV-tk/GCV基因治疗系统杀伤ACC-M

Objective To understand the mechanism of cell death induced by HSV tk/GCV system.Methods Herpes simplex virus thymidine kinase gene (HSV tk) was transferred into human adenoid cystic carcinoma cell strain ACC M cells with lipofect AMINE,the death of gene transduced cells was observed with fluorescent microscopy,DNA electrophoresis and flow cytometry(FCM) after adding different concentrations of GCV at different time.Results There were apparent apoptotic body and chromatin condensation under fluorescent microscope,but DNA ladder only appeared at higher GCV concentration (400 μmol/L) gruop, and FCM found that there was sub G1 peak at each group,the apoptosis rate was 52.87% after 200 μmol/L GCV treatment for 4 days. Conclusion HSV tk/GCV system can kill ACC M cells efficiently and one of the mechanisms is to induce ACC M cell apoptosis.$$$$