- Volume 14 Issue 11
DOI QR Code
Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer
- Xu, Rong (Institute of Molecular Medicine, Huaqiao University) ;
- Guo, Long-Jiang (Institute of Molecular Medicine, Huaqiao University) ;
- Xin, Jun (Department of Urology, the First Hospital of Quanzhou Affiliated Fujian Medical University) ;
- Li, Wen-Mao (Institute of Molecular Medicine, Huaqiao University) ;
- Gao, Yan (Institute of Molecular Medicine, Huaqiao University) ;
- Zheng, You-Xian (Quanzhou Center for Disease Control and Prevention) ;
- Guo, You-Hong (Department of Pharmacy, Quanzhou Medical College) ;
- Lin, Yang-Jun (Department of Pharmacy, Quanzhou Medical College) ;
- Xie, Yong-Hua (Department of Pharmacy, Quanzhou Medical College) ;
- Wu, Ya-Qing (Institute of Molecular Medicine, Huaqiao University) ;
- Xu, Rui-An (Institute of Molecular Medicine, Huaqiao University)
- Published : 2013.11.30
Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.
Luciferase assay;lung cancer;tumor-specific promoter;gene therapy;tumor biomarker
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