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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 细胞的机理之一是诱导细胞凋亡
Objective To explore the mechanism of HSV-tk / GCV (herpes simplex virus thymidine kinase gene / hydroxymethyl acyclovir) gene therapy system to kill human highly metastatic adenoid cystic carcinoma cell line ACC-M. Methods Fluorescent staining fluorescence microscopy, cell DNA electrophoresis, flow cytometry and other methods. Results The results showed that apoptotic bodies and chromatin polyglutination were observed by fluorescence staining. DNA ladder was observed when GCV concentration was high (400 μmol / L) for a long time (4 days) FCM showed a very obvious sub-G1 peak under the effect of 400 μmol / LGCV. Conclusions One of the mechanisms of HSV-tk / GCV gene therapy system killing ACC-M cells is to induce apoptosis