Construction and In Vitro Evaluation of Tissue-Specific Prostate-Specific Antigen Promoter/Enhancer-Regulated Vectors

摘要

To construct specific vectors and evaluate the biological feature of prostate specific antigen (PSA) promoter/enhancer, three fragments harboring PSA enhancer (PSAE) and promoter (PSAP) were amplified using the PCR method. Two PSA promoters, PSA enhancer and pTAL-Luc, were used to construct PSAP1-Luc, PSAP2-Luc, and PSAE-Luc. These recombinant plasmids were co-transfected with a β-galactosidase plas-mid into the three cell lines of PC-3, Hela, and MCF-7. The expression levels of β-galactosidase luciferase were respectively measured quantitatively. The recombinant plasmids, PSAP1-Luc, PSAP2-Luc, and PSAE-Luc, were successfully constructed. Prostate cancer cell PC-3 expressed a higher luciferase activity than that of cervical-cancer cell Hela and breast cancer cell MCF-7 after being transfected with one of the three plasmids. Among the three plasmids in the three types of cells studied, the highest luciferase level was observed with the PSAP2-Luc transfection. The PSA promoters and enhancer possessed some tissue-specific activity in reporter gene analysis. Among them, the PSA promoter2 containing 620 bp showed the highest expression efficiency. In the three cell lines tested, the highest tissue-specific activity of luciferase over β-galactosidase was observed in the PC-3 cells. These relatively tissue-specific promoters have an impact in the gene therapy of prostate cancer.