外源性Muc1、Survivin和Nanog基因启动子在结肠癌干细胞中的表达活性  

Expression of exogenous Muc1,Survivin and Nanog gene promoters in colon cancer stem cells

作　　者：杜敏[1] 葛海燕[2] 徐彬[1] 

机构地区：[1]同济大学附属第十人民医院普外科,上海200072 [2]同济大学附属东方医院普外科,上海200120

出　　处：《第二军医大学学报》2012年第10期1082-1085,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81001007);同济大学附属第十人民医院优秀青年基金(10RQ105)~~

摘　　要：目的探讨结肠癌干细胞中不同基因启动子的活性,寻找可能用于结肠癌干细胞靶向治疗的启动子。方法应用流式细胞仪分选人结肠癌HCT116细胞系中CD133+CD44+标记的肿瘤干细胞。采用PCR技术获取Nanog、Muc1和Survivin基因启动子并分别克隆入pGL3-basic质粒。将含有启动子的pGL3-basic质粒和pGL3-control质粒分别与pRL-sv40共转染结肠癌细胞(HCT116、SW620、HT29)、结肠癌干细胞(CD133+CD44+HCT116)及人正常肝细胞(QSG7701),通过检测双荧光素酶活性以测定启动子活性。结果pGL3-basic-Nanog、pGL3-basic-Muc1、pGL3-basic-Survivin经测序鉴定正确。HCT116细胞系中CD133+CD44+细胞比率约占42.2%。双荧光素酶活性检测显示:在HCT116、SW620、HT29和CD133+CD44+HCT116细胞中,Muc1与Survivin均表现出了较强的活性,而在正常细胞QSG7701中活性较低。结论 Muc1、Survivin启动子可能是用于靶向结肠癌干细胞治疗的有效候选启动子。Objective To study the activities of different gene promoters in colon cancer stem cells and to search for effective promoter for targeted therapy of colon cancer stem cells.Methods CD133＋CD44＋ cells were sorted from cell line HCT116 by flow cytometry.Nanog,Muc1 and Survivin gene promoters were amplified by PCR and were separately cloned into pGL3-basic plasmid.pGL3-basic-promoter plasmid or pGL3-basic-control plasmid together with plasmid pRL-sv40 were co-transfected into colon cancer cell lines（HCT116,SW620,and HT29）,CD133＋CD44＋ HCT116 cells,and normal human liver cell line QSG7701.And then promoter activity was examined by dual-luciferase assays.Results The constructed pGL3-basic-Nanog,pGL3-basic-Muc1 and pGL3-basic-Survivin were confirmed correct by sequencing.We found that 42.2% of HCT116 cells were CD133^＋CD44^＋ cells.Dual-luciferase activity detection showed that Muc1 and Survivin promoters had strong activities in both colon cancer cells and CD133^＋CD44^＋ HCT116 cells,and they had low activities in normal cells QSG7701.Conclusion Survivin and Muc1 promoters may serve as effective candidate for targeted treatment of colon cancer stem cells.

关 键 词：MUC1 NANOG SURVIVIN 启动子 结肠肿瘤干细胞 肿瘤 

分 类 号：R735.35[医药卫生—肿瘤]