Abstract
Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim ofour 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 sevenpromoters, 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, SERPINB3and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found thatthe order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELPand 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 tumourspecific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential foruse in lung cancer targeted gene therapy research.
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