| 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。 |
| 关键词: Muc1 Nanog Survivin 启动子 结肠肿瘤干细胞；肿瘤 |
| DOI：10.3724/SP.J.1008.2012.001082 |
| 投稿时间：2012-02-23修订日期：2012-09-12 |
| 基金项目: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). |
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| Expression of exogenous Muc1, Survivin and Nanog gene promoters in colon cancer stem cells |
| DU Min1,GE Hai-yan2,XU Bin1* |
(1. Department of General Surgery, The Tenth People’s Hospital of Shanghai, Tongji University, Shanghai 200072, China
2. Department of General Surgery, East Hospital, Tongji University, Shanghai 200120, China
*Corresponding author.) |
| Abstract: |
| ObjectiveTo 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. MethodsCD133+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. ResultsThe 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. ConclusionSurvivin and Muc1 promoters may serve as effective candidate for targeted treatment of colon cancer stem cells. |
| Key words: Muc1 Nanog Survivin promoter colonic neoplasms neoplastic stem cells |
