Activation of two mutant androgen receptors from human prostatic carcinoma by adrenal androgens and metabolic derivatives of testosterone.
The androgen receptor (AR) plays a central regulatory role in prostatic carcinoma and is a target of androgen ablation therapy. Recent detection of mutant receptors in tumor specimens suggest a contribution of AR alterations to progression towards androgen independence. In a specimen derived from metastatic prostate cancer we have reported a point mutation in the AR gene that leads to a single amino acid exchange in the ligand binding domain of the receptor. Another amino acid exchange resulting from a point mutation was also identified 15 amino acids away from our mutation. This mutation was detected in the AR gene isolated from an organ-confined prostatic tumor. Here we report the functional characterization of the two mutant receptors in the presence of adrenal androgens and testosterone metabolites. These studies were performed by cotransfecting androgen-responsive reporter genes and either the wild-type or mutant AR expression vectors into receptor negative DU-145 and CV-1 cells. The indicator genes used consisted of the promoter of the androgen-inducible prostate-specific antigen gene or the C' Delta9 enhancer fragment from the promoter of the mouse sex-limited protein driving the expression of the bacterial chloramphenicol acetyl transferase gene. Cotransfection-transactivation assays revealed that the adrenal androgen androstenedione and two products of testosterone metabolism, androsterone and androstandiol, induced reporter gene activity more efficiently in the presence of the mutant receptors than in the presence of the wild-type receptor. No difference between wild-type and mutant receptors was observed in the presence of the metabolite androstandione. The interaction of receptor-hormone complexes with target DNA was studied in vitro by electrophoretic mobility shift assays (EMSA). Dihydrotestosterone and the synthetic androgen mibolerone induced a faster migrating complex with all receptors, whereas the androgen metabolite androstandione induced this complex only with the two mutant receptors. Androsterone and androstandiol were inactive in the EMSA. These aberrant properties of the mutant receptors in the presence of adrenal androgens and products of androgen metabolism may be of importance in the course of the prostate cancer, especially during androgen ablation therapy.
Department of Urology, University of Innsbruck, Austria.
Z
Z
Culig
J
J
Stober
A
A
Gast
H
H
Peterziel
A
A
Hobisch
C
C
Radmayr
A
A
Hittmair
G
G
Bartsch
A C
AC
Cato
H
H
Klocker
D000728:Androgens / Q000494:pharmacology*; D000735:Androstenedione / Q000494:pharmacology; D000738:Androsterone / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics / Q000502:physiology; D003181:Complement C4; D004273:DNA, Neoplasm / Q000378:metabolism; D000882:Haplorhini; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D017430:Prostate-Specific Antigen / Q000235:genetics / Q000502:physiology; D011471:Prostatic Neoplasms / Q000235:genetics / Q000648:ultrastructure*; D011944:Receptors, Androgen / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D013739:Testosterone / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D015533:Transcriptional Activation; D014162:Transfection
D000728:Androgens; D001798:Blood Proteins; C018386:C4a protein, mouse; D003181:Complement C4; D004273:DNA, Neoplasm; D011944:Receptors, Androgen; D013739:Testosterone; D000735:Androstenedione; D000738:Androsterone; D017430:Prostate-Specific Antigen