全反式维甲酸增强HSV-tk/GCV系统对晶状体上皮细胞的旁观者效应

All-trans retinoic acid enhances bystander effect of HSV-tk/GCV system in rabbit lens epithelial cells in vitro

目的:体外观察兔晶状体上皮细胞N/N1003A经全反式维甲酸(all-trans retinoic acid,ATRA)诱导后,2型重组腺相关病毒(recombinant adeno-associated virus2,rAAV2)介导的单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine kinase,HSV-tk)/丙氧鸟苷(ganciclovir,GCV)自杀基因系统对细胞旁观者效应的影响。方法:应用RT-PCR法检测rAAV2介导HSV-tk基因转染N/N1003A细胞后TK基因的表达,MTT法观察经ATRA诱导后HSV-tk/GCV系统对N/N1003A细胞旁观者效应的变化,流式细胞术检测经ATRA诱导后HSV-tk/GCV系统对N/N1003A细胞凋亡的影响。结果:rAAV2介导HSV-tk基因成功转染N/N1003A细胞并稳定表达。与对照组比较,ATRA诱导组可显著提高HSV-tk/GCV系统对兔晶状体上皮细胞N/N1003A的旁观者效应(P<0.01),且ATRA诱导组细胞凋亡率显著增加(P<0.01)。结论:ATRA可显著增强rAAV2介导的HSV-tk/GCV系统对体外培养兔晶状体上皮细胞的旁观者效应。

AIM： To investigate the effect of all-trans retinoic acid（ATRA）on the bystander effect of herpes simplex virus thymidine kinase（HSV-tk）/ganciclovir（GCV）suicide gene system mediated by recombinant adeno-associated virus 2（rAAV2）in lens epithelial cells（N/N1003A）in vitro.METHODS： The expression of TK gene in lens epithelial cells mediated by rAAV2/HSV-tk was detected by RT-PCR. MTT assay was used to observe the changed bystander effects of HSV-tk/GCV system in N/N1003A cells after ATRA therapy. The effects of ATRA therapy on the apoptotic rates of HSV-tk/GCV system in N/N1003A cells were investigated by flow cytometry.RESULTS： HSV-tk gene mediated by rAAV2 was transfected successfully into N/N1003A cells and expressed stably. The bystander effect of HSV-tk/GCV system on N/N1003A cells in ATRA treatment group was significantly improved when compared with control group（P〈0.01）, and the apoptotic rates of the study group were notably increased（P〈0.01）.CONCLUSION： ATRA can significantly enhance the bystander effect of HSV-tk/GCV suicide gene system mediated by rAAV2 in rabbit lens epithelial cells in vitro.