环丙氧苷诱导转导有外源HSV—tk基因的C6胶质瘤细胞的 …

目的 观察单纯疱疹病毒胸苷激酶基因（ＨＳＶ－ｔｋ）－环丙氧苷（ＧＣＶ）系统在胶质瘤基因治疗过程中,ＧＣＶ能否诱导转导有ＨＳＶ－ｔｋ基因的胶质瘤细胞凋亡。方法 采用光镜、电镜和荧光显微镜观察转导有ＨＳＶ－ｔｋ基因的大鼠Ｃ６／ｔｋ胶质瘤细胞形态学的改变,以末转导ＨＳＶ－ｔｋ基因的Ｃ６细胞作为对照,并以流式细胞术,ＤＮＡ凝胶电流分析进一步验证细胞凋亡的发生。结果 在５μｇ／ｍｌ ＧＣＶ作用７２ｈ后,Ｃ６

Ganciclovir induces apoptosis of rat C6 glioma cell transduced with HSV-tk gene

OBJECTIVE: To observe whether apoptosis could be induced by ganciclovir (GCV) in the herpes simplex virus-thymidine kinase gene-ganciclovir system (HSV-tk-GCV system) gene therapy for glioma. METHODS: Transduced C6/tk glioma cell as treated group, wild C6 cell as control group, were studied with morphology, gel-electrophoresis of DNA fragment analysis, fluoroscopy, and flow cytometric study before or after exposed to GCV, and apoptosis of transduced C6/tk glioma cell was ascertained. RESULTS: When C6/tk cell was treated with GCV at 5 micrograms/ml for 72 hours, cell apoptosis occurred. Typical apoptotic morphological features included cell shrinkage, condensation, and margination of nuclear chromatin were showed by light and electron microscopy, condensed nuclear chromatin and the fragment of nuclei were demonstrated by fluoroscopy, DNA ladder was showed by DNA fragment analysis, apoptotic peak was identified by flow cytometric study. The apoptotic cells accounted for 23% of the cell population. Apoptosis was not observed in control group treated by GCV. CONCLUSIONS: The apoptosis of transduced C6/tk glioma was induced by GCV, which fit in with the basis of HSV-tk-GCV gene therapy for glioma.