Development of prostate-specific antigen promoter-based gene therapy for androgen-independent human prostate cancer.
OBJECTIVE
The goal of this study is to develop a tissue-specific toxic gene therapy utilizing the prostate specific antigen (PSA) promoter for both androgen-dependent (AD) and androgen-independent (AI) PSA-secreting prostate cancer cells. Ideally this gene therapy would be effective without the necessity of exposing the target cells to circulating androgens.


METHODS
An AI subline of LNCaP, an AD PSA-secreting human prostate cancer cell line, C4-2, was used in this study. Castrated mice bearing C4-2 tumors secrete PSA. A transient expression experiment was used to analyze the activity of two PSA promoters, a 5837 bp long PSA promoter and a 642 bp short PSA promoter, in C4-2 cells. A recombinant adenovirus (Ad-PSA-TK) carrying thymidine kinase under control of the long PSA promoter was generated. The tissue-specific activity of Ad-PSA-TK was tested in vitro and in vivo.


RESULTS
The long PSA promoter had superior activity over short PSA promoter, and higher activity in C4-2 cells than in LNCaP cells. High activity of Ad-PSA-TK was observed in C4-2 cells in an androgen deprived condition. In vitro, Ad-PSA-TK was further demonstrated to induce marked C4-2 cell-kill by acyclovir in medium containing 5% FBS. No cell-kill was observed in control WH cells (a human bladder cancer cell line). In vivo, Ad-PSA-P-TK with acyclovir significantly inhibited subcutaneous C4-2 tumor growth and PSA production in castrated animals.


CONCLUSIONS
The 5837 bp long PSA promoter was active in the androgen free environment and could be used to target both androgen-dependent and independent PSA-producing prostate cancer cells in vitro, and prostate tumors in castrated hosts.
Gotoh
A
A
Department of Urology, Molecular Urology and Therapeutics Program, University of Virginia, Charlottesville 22908, USA.
Ko
S C
SC
Shirakawa
T
T
Cheon
J
J
Kao
C
C
Miyamoto
T
T
Gardner
T A
TA
Ho
L J
LJ
Cleutjens
C B
CB
Trapman
J
J
Graham
F L
FL
Chung
L W
LW
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015316:Genetic Therapy; D006801:Humans; D008297:Male; D051379:Mice; D017430:Prostate-Specific Antigen / Q000096:biosynthesis / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D011995:Recombination, Genetic; D013045:Species Specificity; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D013937:Thymidine Kinase; D017430:Prostate-Specific Antigen