title: Prostate-specific antigen promoter driven gene therapy targeting DNA polymerase-alpha and topoisomerase II alpha in prostate cancer.
abstract: Attainment of cell type-specific cytotoxicity with minimal side effects is the ultimate goal of cancer therapy. By employing the prostate-specific antigen promoter (PSAP), we investigated (1) whether PSAP-driven antisense genetic constructs targeting DNA polymerase-alpha and topoisomerase II alpha (Top II alpha), designated PSAP-antipol and PSAP-antitop respectively, could induce death of prostate cancer cells, and (2) whether the cytotoxicity is restricted to cells of prostate origin. A PSAP-driven beta-galactosidase gene, PSAP-LacZ, was also used to estimate the expression of the PSAP-driven transcripts. Lipofection-mediated gene transfers were performed with these 3 constructs and a control plasmid, pCDNA3, in 3 human prostate cancer cell lines (LNCaP, DU-145, PC-3) and 5 other cell lines (Cos-1 [monkey kidney], HL-60 [human myeloid leukemia], Hep G2 [human hepatoma], NCI H460 [human lung cancer] and SW 480 [human colon cancer]). On transfection with PSAP-LacZ, LNCaP, DU-145, and PC-3 showed a 10.8, 1.8, and 1.6 fold increase in beta-galactosidase activity, respectively. The remaining 5 cell lines showed no changes after transfection. Corresponding to the levels of the induced beta-galactosidase activity, LNCaP showed the strongest growth inhibition by the antisense constructs: 36% by PSAP-antipol, 39% by PSAP-antitop and 80% by PSAP-antipol+PSAP-antitop. DU-145 and PC-3 had minimal growth inhibition with PSAP-antipol alone or PSAP-antitop alone. However, when cotransfected with PSAP-antipol and PSAP-antitop, DU-145 and PC-3 displayed 42% and 55% growth inhibition, respectively. In contrast, no cytotoxicity was observed in the remaining 5 cell lines when transfected with PSAP-antipol, PSAP-antitop or both. Therefore, PSAP-driven antisense gene therapy targeting DNA polymerase-alpha and Top II alpha inhibits the growth of human prostate cancer cells and the cytotoxic effect is restricted in cells of prostate origin.
authors.affiliation: Division of Laboratory Medicine, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
authors.forename: C H
authors.initials: CH
authors.lastname: Lee
authors.forename: M
authors.initials: M
authors.lastname: Liu
authors.forename: K L
authors.initials: KL
authors.lastname: Sie
authors.forename: M S
authors.initials: MS
authors.lastname: Lee
mesh_terms: D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D003110:Colonic Neoplasms; D004257:DNA Polymerase II / Q000096:biosynthesis*; D017931:DNA Primers; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis*; D016373:DNA, Antisense / Q000494:pharmacology*; D018076:DNA, Complementary; D015870:Gene Expression; D015316:Genetic Therapy; D018922:HL-60 Cells; D006801:Humans; D007668:Kidney; D008113:Liver Neoplasms; D008175:Lung Neoplasms; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017430:Prostate-Specific Antigen / Q000096:biosynthesis / Q000235:genetics*; D011471:Prostatic Neoplasms; D011994:Recombinant Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D017931:DNA Primers; D016373:DNA, Antisense; D018076:DNA, Complementary; D011994:Recombinant Proteins; D004257:DNA Polymerase II; D001616:beta-Galactosidase; D017430:Prostate-Specific Antigen; D004250:DNA Topoisomerases, Type II