Gene therapy for prostate cancer by targeting poly(ADP-ribose) polymerase.
Poly(ADP-ribose) polymerase (PARP) has strong affinity for DNA strand breaks and cycles on and off the DNA ends to allow DNA repair. A DNA-binding domain of PARP (PARP-DBD) acts as a dominant-negative mutant by binding to DNA strand breaks irreversibly and sensitizing mammalian cells to DNA-damaging agents. Therefore, expression of PARP-DBD in prostate carcinoma cells offers a strategy to achieve sensitization to genotoxic treatments. Toward this end, we developed recombinant plasmids expressing the PARP-DBD under the control of the 5'-flanking sequences of the human prostate-specific antigen (PSA) gene. Tissue specificity of PARP-DBD expression in human tumor cells was confirmed using the PSA-producing (LNCaP) and PSA-negative (PC-3) prostate cancer cells, as well as cells of nonprostate origin, Ewing's sarcoma (A4573 cells). LNCaP cells stably transfected with the PSA-regulated cDNA for PARP-DBD exhibit an androgen-dependent induction of PARP-DBD expression as determined by Western blotting, reverse transcription-PCR, and in situ immunofluorescence. Furthermore, we found that PARP-DBD sensitized LNCaP cells to DNA-damaging agents, such as ionizing radiation and etoposide. Androgen (R1881) -dependent stimulation of PARP-DBD expression resulted in a 2-fold growth inhibition in LNCaP cells as compared with controls, and an augmented apoptotic cell death in response to ionizing radiation or etoposide. Taken together, the plasmid vector developed in this study permits the expression of the human PARP-DBD in an androgen-inducible and PSA-dependent fashion, and sensitizes prostatic adenocarcinoma cells to DNA-damaging treatments. These results provide proof-of-principle for a novel therapeutic strategy for the treatment of prostate cancer.
Trofimova
Irina
I
Department of Radiation Medicine, Georgetown University Medical Center, Washington, DC 20007, USA.
Dimtchev
Alexandre
A
Jung
Mira
M
Rosenthal
Dean
D
Smulson
Mark
M
Dritschilo
Anatoly
A
Soldatenkov
Viatcheslav
V
D000230:Adenocarcinoma / Q000201:enzymology* / Q000235:genetics / Q000628:therapy*; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D003131:Combined Modality Therapy; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D005047:Etoposide / Q000494:pharmacology; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008297:Male; D015741:Metribolone / Q000494:pharmacology; D009376:Neoplasms, Hormone-Dependent / Q000201:enzymology / Q000235:genetics / Q000628:therapy; D010957:Plasmids / Q000235:genetics; D011065:Poly(ADP-ribose) Polymerases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017430:Prostate-Specific Antigen / Q000096:biosynthesis / Q000235:genetics; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000628:therapy*; D017434:Protein Structure, Tertiary; D045165:Testosterone Congeners / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D000972:Antineoplastic Agents, Phytogenic; D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D045165:Testosterone Congeners; D015741:Metribolone; D005047:Etoposide; D011065:Poly(ADP-ribose) Polymerases; D017430:Prostate-Specific Antigen