摘要
目的探讨HSV-tk/GCV(人类单纯疱疹病毒胸苷激酶联合更昔洛韦)自杀基因治疗系统对喉鳞状细胞癌细胞系Hep-2的体外作用。方法脂质体介导HSV-tk基因转染Hep-2细胞,逆转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)检测HSV-tk基因的表达;四甲基偶氮唑蓝法检测该系统对Hep-2细胞的体外抑制作用,流式细胞仪检测细胞周期阻滞和凋亡现象。结果脂质体能介导HSV-tk基因转染Hep-2细胞;HSV-tk/GCV系统对Hep-2细胞具有生长抑制效应;更昔洛韦(ganciclovir,GCV)浓度为20μg/ml时其抑制率达到40%;HSV-tk/GCV自杀基因治疗系统主要通过S期细胞周期阻滞对Hep-2细胞发挥体外抑制作用,没有明显的细胞凋亡和坏死。结论HSV-tk/GCV系统主要通过细胞周期阻滞作用对Hep-2细胞发挥体外抑制作用。HSV-tk/GCV系统对Hep-2细胞是否具直接杀伤作用有待于进一步探讨。HSV-tk/GCV系统具有潜在临床应用价值。
Objective Efficacy of HSV-tk/GCV system antitumor effects was assessed on human laryngeal cancer cell line Hep-2 in vitro. To assess the HSV-tk/CGV system whether has an antitumour effect on human laryngeal squamons cell cancer Hep-2 in vitro. The mechanisms of cytotoxity were also assessed. Methods Hep-2 ceils were transfected with HSV-tk gene by lipofection . Reverse transcription polymerase chain reaction （RT-PCR） was used to detect the HSV-tk gene expression. MTT was utilized to test for the cytotoxicity of this system. The cell-circle arrest and apoptosis were analyzed by flowcytometery assay. Results HSV-tk gene transfected cells demonstrated obvious cytoreductivity followed by ganciclovir （GCV） administration and this cytoreductivity showed partial GCV dose-independent. HSV-tk gene transfected cells demonstrated obvious s-phase arrest, no apoptosis and necrosis occurred. Condusions The HSV-tk/GCV system can inhabit the growth of Hep-2 cells effectively. S-phase arrest perhaps is the main reason that leads to the cell inhibition in our study. HSV-tk/GCV system has potential antitumor effects for the future clinical practice.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2006年第5期374-376,共3页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
关键词
喉肿瘤
胸苷激酶
基因疗法
Laryngeal neoplasms
Thymidine kinase
Gene therapy
