中文摘要：

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。

英文摘要：

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.