丹参酮ⅡA增强HSV-tk/GCV旁观者效应及其与Cx43 mRNA表达的关系  

Enhancement of the bystander effect by tanshinone Ⅱ A in HSV-tK/GCV system is related to expression of connexin 43 mRNA

作　　者：黄光琦[1] 宋毅[1] 张洁[1] 陆燕蓉[1] 肖林[1] 杨元[2] 郭元彪[3] 

机构地区：[1]四川大学华西医院肿瘤研究所,成都610041 [2]四川大学华西临床医学院医学遗传教研室 [3]四川大学华西基础医学院生化教研室

出　　处：《中华肿瘤杂志》2004年第3期146-149,共4页Chinese Journal of Oncology

摘　　要：目的　研究丹参酮ⅡA(TanshinoneⅡA ,Tan)对HSV tk/GCV旁观者效应的增强作用及其与间隙连接蛋白Cx4 3转录表达的关系。方法　应用polybrene转染、荧光定量RT PCR等技术 ,观察Tan对宫颈癌细胞ME180 (ME)、ME/TK转化细胞旁观者效应及诱导Cx4 3mRNA表达的作用。结果　在HSV tk/GCV系统中 ,Tan显著地提高了ME/TK细胞对GCV的敏感性。在 2 μg/mlGCV作用下 ,以1.3× 10 9mol/LTan与不加Tan的作用相比较 ,在ME与ME/TK不同比例混合细胞的各组中 ,细胞的存活率明显降低 ,差异均有显著性 (P <0 .0 5 )。并观察到Tan对GCV旁观者效应的增强作用 ,在一定范围内为 1.3× 10 -8mol/L和 1.3× 10 -9mol/L。RT PCR结果表明 ,经 1.3× 10 -8mol/L和 1.3× 10 -9mol/LTan处理的ME细胞 ,其Cx4 3mRNA的相对拷贝数比值增高约 8.83倍及 8.4 7倍。结论　在宫颈癌ME180细胞中 ,Tan在 1.3× 10 -8mol/L和 1.3× 10 -9mol/L范围内具有明显增强HSV tk/GCV旁观者效应的作用。Tan在转录水平诱导Cx4 3mRNA表达上调 ,与旁观者效应的增强作用密切相关。ObjectiveTo investigate enhancement of the bystander effect by tanshinone ⅡA(Tan) in HSV-tK/GCV system and the correlation with expression of connexin 43 mRNA. MethodsThe cytotoxic effect in HSV-tK/GCV in cervical carcinoma cell line ME180 (ME) and ME/TK was examined by MTT assays. Cx43 mRNA expression was detected by fluor-quantitative RT-PCR. ResultsTan markedly increased sensitivity of ME/TK cells for GCV in HSV-tK/GCV system. In the presence of 2 μg/ml GCV, compared with the absence of Tan (0 mol/L) , an obvious decrease in survival rate was seen at any given mixture of ME and ME/TK cells exposed to 1.3×10 -9 mol/L Tan. Statistics showed significant difference( P <0.05). However, enhancement of bystander mediated cell killing occurred only in the range of Tan concentrations used (1.3×10 -8 , 1.3×10 -9 mol/L). RT-PCR showed that the ratio of relative copy number of Cx43 mRNA increased by 8.83 and 8.47 -fold in ME cells exposed to 1.3× 10 -8 and 1.3×10 -9 mol/L Tan, respectively. ConclusionFor the first time we report that in cervical carcinoma ME180 cell line, Tan possesses a remarkable enhancing role on the bystander effect in the HSV-tK/GCV system. It is associated with up-regulation of Cx43 mRNA expression.

关 键 词：丹参酮ⅡA 旁观者效应 CX43 靼纯疱疹病毒-胸苷激酶基因 丙氧基鸟苷 自杀基因 基因治疗 

分 类 号：R73-36[医药卫生—肿瘤]