Expression of exogenous mud, survivin and nanog gene promoters in colon cancer stem cells
10.3724/SP.J.1008.2012.01082
- Author:
Du MIN
1
Author Information
1. Department of General Surgery, The Tenth People's Hospital of Shanghai, Tongii University
- Publication Type:Journal Article
- Keywords:
Colonic neoplasms;
Md1;
Nanog;
Neoplastic stemcells;
Promoter;
Survivin
- From:
Academic Journal of Second Military Medical University
2012;33(10):1082-1085
- CountryChina
- Language:Chinese
-
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.
