Expression of exogenous mud, survivin and nanog gene promoters in colon cancer stem cells / 第二军医大学学报

ABSTRACT

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, Mud 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 activitywas examined by dual-luciferase assays. Results The constructed pGL3-basic-Nanog, pGL3-basic-Md1 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 Mud 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 Mud promoters may serve as effective candidate for targeted treatment of colon cancer stem cells.