A33启动子结肠癌特异性探究及SV40增强子对其转录活性影响  

Investigation of A33 Promoter's Specifically Transcriptional Activity in Colorectal Cancer and the Influence When Combined with SV40 Enhancer

作　　者：钟丹[1] 郑水娣[1] 谌贺宽子[1] 袁素敬[1] 章康健[2] 

机构地区：[1]浙江理工大学生命科学学院新元医学与生物技术研究所,杭州310018 [2]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《中国细胞生物学学报》2011年第11期1241-1247,共7页Chinese Journal of Cell Biology

基　　金：浙江理工大学基金(No.1016819-Y)资助项目~~

摘　　要：为了探究A33核心启动子结肠癌特异性及SV40增强子对其转录水平的影响,该研究通过构建A33核心启动子和带SV40增强子的A33核心启动子(eA33)的荧光素酶报告基因载体pGL3-A33和pGL3-eA33,与内参照pRL-SV40质粒共转染至不同的细胞系中,利用双荧光素酶检测系统检测分析了A33和eA33启动子在不同细胞系中的转录活性。结果显示,A33核心启动子在结肠癌细胞系中具有转录活性低,但结肠癌特异性好的特点,而在其他类型癌细胞中基本没有活性。同时发现,eA33在各类癌细胞中的转录水平与A33相比,均呈大幅度提高,有显著性差异(P<0.01),但SV40增强子能显著增强A33启动子转录活性的同时减低了其结肠癌特异性。这为靶向癌症基因—病毒治疗策略在结肠癌的生物治疗应用中寻找结肠癌特异性的启动子奠定了研究基础。In order to investigate A33 core promoter＇s specifically transcriptional activity in colorectal cancer （CRC） and to identify its transcriptional activity combined with SV40 enhancer, the transcriptional regula- tory elements were constructed by appending a SV40 enhancer 5＇ to the A33 cOre promoter （eA33）, and then we constructed two luciferase reporter gene vectors which containing A33 promoter and eA33 promoter, pGL3-A33 and pGL3-eA33, and made them co-transfected with transfection-efficiency normalization vector pRL-SV40 into different cell lines by liposome transfection. The transcriptional activity of A33 and eA33 promoter was analyzed by Dual-Luciferase Assay system and relative luciferase unit （RLU） was used to evaluate the expression efficiency. The results showed that A33 promoter performs high CRC specific expression but low transcriptional activity, and showed barely any activity in other types of cancer cells. Meanwhile, the eA33 promoter＇s transcriptional activity is slightly stronger than the A33 promoter＇s in all kinds of cell lines. Therefore, the SV40 enhancer is able to greatly up-regulate A33 promoter transcriptional activity, and by the meantime, attenuate the CRC specific of A33 promoter. It will definitely provide useful information for targeted gene-viro-therapy of CRC.

关 键 词：A33核心启动子 SV40增强子 结肠癌特异性 基因-病毒治疗 

分 类 号：R735.35[医药卫生—肿瘤]