title: Recombinant adenovirus mediated prostate-specific enzyme pro-drug gene therapy regulated by prostate-specific membrane antigen (PSMA) enhancer/promoter.
abstract: Gene directed enzyme pro-drug therapy (GDEPT) is one of the adjuvant therapeutic regimens for advanced prostate adenocarcinoma, and this research intended to explore how to apply targeting therapy of prostate adenocarcinoma under the mediation of a promoter/enhancer of prostate-specific membrane antigen (PSMA(EP)) as a specific regulatory element. Recombinant adenoviruses (Ad-PSMA(E-P)-enhanced green fluorescent protein [EGFP], Ad-CMV-EGFP, Ad-PSMA(E-P)-CD, and Ad-CMV-CD) were constructed and could express cytosine deaminase (CD) or the EGFP reporter gene driven by a PSMA(EP) or cytomegalovirus (CMV) promoter. LNCaP, CL-1, MCF-7, and A549 were infected with CD-produced recombinant adenoviruses and treated with pro-drug 5-fluorocytosine (5-FC) in vivo and vitro; then, the growth inhibition of the cells and the cell cycle variation were assessed by an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assay and flow cytometry. Growth suppression of the xenograft tumor was also adopted to evaluate the efficiency of the suicide system. Morphologic changes after treatment in vivo were assessed with hematoxylin and eosin staining. In the 4 examined cancer cell lines, PSMA-positive prostate cancer cells LNCap and CL-1 were exclusively sensitive to the Ad-PSMA(E-P)-CD/5-FC system. The S phase of cell cycle arrest was thought to be involved in the cytotoxicity of 5-fluorouracil (5-FU) converted from 5-FC by CD. CL-1 implanted Athymic BALB/c mice showed growth inhibition of tumors when they were treated with the Ad-PSMA(E-P)-CD/5-FC system without systemic conversion toxicity. The PSMA-based, CD-produced adenovirus, deserving further investigation in the future, might be a good candidate for targeting gene therapy of prostate adenocarcinoma.
authors.affiliation: Department of Urology, West China Hospital, Sichuan University, 37 Guoxue Xiang St, Chendu, Sichuan, People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zeng
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.forename: Rui
authors.initials: R
authors.lastname: Huang
authors.forename: Ni
authors.initials: N
authors.lastname: Chen
authors.forename: Qiang
authors.initials: Q
authors.lastname: Dong
authors.forename: Yuru
authors.initials: Y
authors.lastname: Yang
authors.forename: Qiao
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics* / Q000494:pharmacology / Q000633:toxicity; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003587:Cytomegalovirus / Q000235:genetics; D004742:Enhancer Elements, Genetic; D017930:Genes, Reporter; D015316:Genetic Therapy; D043425:Glutamate Carboxypeptidase II / Q000235:genetics* / Q000494:pharmacology / Q000633:toxicity; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms; D011995:Recombination, Genetic; D014158:Transcription, Genetic
chemical_list: D000954:Antigens, Surface; D049452:Green Fluorescent Proteins; C473848:FOLH1 protein, human; D043425:Glutamate Carboxypeptidase II