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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 To investigate the in vitro effects of HSV-tk / GCV (human herpes simplex virus thymidine kinase plus ganciclovir) suicide gene therapy system on laryngeal squamous cell carcinoma cell line Hep-2. Methods HSV-tk gene was transfected into Hep-2 cells with lipofectamine 2000. The expression of HSV-tk gene was detected by reverse transcription polymerase chain reaction (RT-PCR) The inhibitory effect of this system on Hep-2 cells was detected by flow cytometry, and the cell cycle arrest and apoptosis were detected by flow cytometry. Results The HSV-tk gene was transfected into Hep-2 cells by liposomes. HSV-tk / GCV system had a growth inhibitory effect on Hep-2 cells. When ganciclovir (GCV) was 20μg / ml, Rate of 40%; HSV-tk / GCV suicide gene therapy system mainly through the S phase cell cycle arrest Hep-2 cells in vitro inhibition, no significant apoptosis and necrosis. Conclusion HSV-tk / GCV system can inhibit Hep-2 cells in vitro through cell cycle arrest. Whether HSV-tk / GCV system has a direct killing effect on Hep-2 cells remains to be further explored. HSV-tk / GCV system has potential clinical value.