全反式维甲酸提高HSV-TK/GCV系统治疗舌鳞癌旁观者效应的研究

All-trans retinoic acid augments the bystander effect of herpes simplex virus thymidine kinase/ganciclovir system in the treatment of tongue carcinoma cell line

目的 探讨全反式维甲酸 (all transretinoicacid ,ATRA)对单纯疱疹病毒胸苷酸激酶(herpessimplexvirusthymidinekinase ,HSV TK) /丙氧鸟苷 (ganciclovir,GCV)系统治疗舌鳞癌旁观者效应的增强作用及机制。方法 应用免疫细胞化学、流式细胞仪等技术 ,对ATRA诱导舌鳞癌细胞系(Tca81 1 3)细胞间隙连接蛋白基因Cx43的表达进行分析 ;采用析因分析法分析ATRA对HSV TK/GCV系统旁观者效应的影响。结果 舌鳞癌细胞经 1 0 - 7mol/L～ 1 0 - 5mol/LATRA处理后Cx43蛋白的表达明显提高 ;阳性细胞计数率由处理前的 5 1 7%上升至 30 53 % (1 0 - 5mol/LATRA) ,经t检验处理组与非处理组间差异有显著性 (P <0 0 1 ) ;舌鳞癌细胞经ATRA处理后增强了HSV TK/GCV系统的旁观者效应 ,两者间存在交互作用 (P <0 0 5)。结论 ATRA可显著提高HSV TK/GCV系统的旁观者效应 。

Objective To investigate the augmentation effect and mechanism of all trans retinoic acid(ATRA)on the bystander effect of herpes simplex virus thymidine kinase (HSV TK)/ganciclovir(GCV) system in the treatment of tongue carcinoma cell line. Methods Immunocytochemistry and flow cytometry (FCM) were used to analyze the expression of Cx43 protein in Tca8113 cells after treated with ATRA; The bystander effect of HSV TK/GCV system on tongue carcinoma cells before and after treatment with ATRA was detected by MTT assays. The interaction of ATRA and bystander effect was analyzed by factorial experiment. Results After treated by ATRA (10 -7 mol/L～10 -5 mol/L), the expression of Cx43 protein was up regulated in Tca8113 cells and the positive rate of Cx43 protein increased from 5 17% (before treatment) to 30.53% (10 -5 mol/L ATRA). There was significant difference between ATRA treated cells and untreated cells( P < 0.01). The bystander effect of HSV TK/GCV system on Tca8113 cells was poor, but improved after combined with ATRA. There was cooperation effect between ATRA and bystander effect of HSV TK/GCV ( P < 0.05). Conclusions ATRA can augment the bystander effect of HSV TK/GCV system in the treatment of tongue carcinoma cell line. The mechanism might be due to up regu1ation of Cx43 gene and restore gap junction intercellular communication.