The Effects Of CD&UPRT/5-FC Double Suicide Gene System On Prostate Cancer Cell Was Improved By The Targeted Regulation Of Prostate-specific Membrane Antigen Promoter And Enhancer

Objective To construct the recombinant plasmid pPSMApromoter /enhancer-CD (pPSMA_E/P-CD) which Cytosine Deaminase(CD) gene is regulated by PSMA promoter/enhancer. To explore the specially targeted killing role of the pPSMA_E/P-CD & Uracil Phosphoribosyltransferase(UPRT) /5-Flurocytosine(5-FC) double suicide gene system to prostate cancer. Methods â‘ CD gene is amplified from E.coli JM109 by polymerase chain reaction (PCR). CD gene was cloned into the recombinant plasmid, to construct recombinant plasmid pPSMA_E/P-CD regulated by PSMA promoter/enhancer, and the recombinant plasmids pCMV-CD and pCMV-UPRT regulated by cytomegalovirus (CMV). Each recombinant plasmid has been verified by PCR , digestion and DNA sequencing. â‘¡After pPSMA_E/P-EGFP contained reportor gene EGFP was transfected into prostate cancer cell line and non-prostate cancer cell line by Lipofectamine 2000, EGFP expression was exclusively observed in LNCaP cell line, which indicated the specific regulatory function of PSMA_E/P. â‘¢ pPSMAe/p-CD, pPSMA_E/P-UPR, pCMV-CD, pCMV-UPRT, pPSMA_E/P-CD&UPRT...