Article Figures & Data
Figures
- Fig. 1.
Concentration of dihydrotestosterone (DHT) and androstenediol (adiol) in benign prostate hypertrophy (BPH) and prostate cancer (PCa) tissue after hormone therapy. Fourteen BPH tissues from patients who did not undergo hormone therapy and 12 PCa tissues from patients who were subjected to neoadjuvant hormone therapy were collected by open surgery. The concentrations of DHT and adiol were measured with high-performance liquid chromatography. The data are presented as the mean of triplicate measurements; bars, ± SE.
- Fig. 2.
Effect of androstenediol (adiol) on LNCaP cells. A, 24 h after 1 × 105 LNCaP cells were seeded, cells were treated with the indicated concentration of adiol, dihydrotestosterone (DHT), testosterone, androstenedione (4-dione), or dehydroepiandrosterone (DHEA) and cultured for 4 days. The cells then were counted by a hemocytometer. The medium was changed every 2 days, and the reagents were added at the same time. B, regulation of prostate-specific antigen (PSA) mRNA by androgens. Twenty-four h after LNCaP cells were seeded, cells were treated with the indicated concentrations of adiol, DHT, 4-dione, or DHEA and cultured for 12 h. The cells then were harvested, and total RNA was extracted. Reverse transcription-PCR analysis of PSA mRNA was performed according to the “Materials and Methods.” C, the amplified products from PSA mRNA in B were quantitated by NIH image 1.61 and normalized by GAPDH. The data are presented as the mean of triplicate experiments; bars, ±SD.
- Fig. 3.
Effect of androgens on prostate-specific antigen (PSA) promoter in LNCaP cells. A, 2 × 105 LNCaP cells were transfected with 0.5 μg pGLPSAp5.8, 1.5 μg pBluescript, and 0.2 μg β-gal expression plasmid. Twenty-four h after transfection, cells were treated with the indicated concentrations of androstenediol (adiol), dihydrotestosterone (DHT), testosterone, androstenedione (4-dione), or dehydroepiandrosterone (DHEA) for 24 h, and luciferase activities then were measured. The data were normalized with β-galactosidase activity. B, effect of bicalutamide on adiol activity in LNCaP cells. The 2 × 105 LNCaP cells were transfected with 0.5 μg pGLPSAp5.8, 1.5 μg pBluescript, and 0.2 μg β-gal expression plasmid. Twenty-four h after transfection, cells were treated with 10−9 m adiol or DHT and the indicated concentrations of bicalutamide for 24 h, and luciferase activities then were measured. The data were normalized with β-galactosidase activity.
- Fig. 4.
Effect of androstenediol (adiol) on PC-3 cells transfected with wild androgen receptor (AR). The 2 × 105 PC-3 cells were transfected with 0.5 μg pGLPSAp5.8, 1.0 μg pCMV-hAR, 0.5 μg pBluescript, and 0.2 μg β-gal expression plasmid. Twenty-four h after transfection, cells were treated with the indicated concentrations of adiol, dihydrotestosterone (DHT), androstenedione (4-dione), or dehydroepiandrosterone (DHEA) for 24 h, and luciferase activities then were measured. The data were normalized with β-galactosidase activity. The data are presented as the mean of triplicate experiments; bars, ±SD.
- Fig. 5.
Effect of androgen receptor (AR) mutation and ARA70 on androstenediol (adiol) activity in PC-3 cells. The 1 × 105 PC-3 cells were transfected with 0.5 μg pGLPSAp5.8, 0.25 μg pCMV-hAR or pCMV-hARmut877, 1.25 μg pSG5 or pSG5ARA70, and 0.2 μg β-gal expression plasmid. Twenty-four h after transfection, cells were treated with the indicated concentrations of 10−10 m adiol or dihydrotestosterone (DHT) for 24 h, and luciferase activities then were measured. The data were normalized by β-galactosidase activity. The data are presented as the mean of three experiments; bars, ±SD.
- Fig. 6.
Scatchard analysis of dihydrotestosterone (DHT) and androstenediol (adiol) binding to androgen receptor (AR) in LNCaP cells (not the wild-type AR). A, [3H]DHT binding to mutant AR (Kd = 0.58 nm). B, [3H]adiol binding to mutant AR (Kd = 1.90 nm)
- Fig. 7.
Nuclear androgen receptor (AR) expression in PC-3 cells transfected with AR and LNCaP cells. Nuclear proteins were extracted as described in “Materials and Methods” and loaded on an 8% SDS-polyacrylamide gel for Western blot analysis. After protein was transferred to polyvinylidene diflouride membrane, and NH27 anti-AR antibody was used for detection of Mr 110,000 AR. Band density was measured by NIH image 1.62. The data are presented as mean. A, 7 × 105 PC-3 cells were transfected with 4 μg pCMV-hAR or pCMV-hARmut877 as described in Fig. 4 <$REFLINK> . One day after transfection, cells were cultured in the absence (c) or presence of 1 × 10−9 m dihydrotestosterone (DHT) (d) or androstenediol (adiol) (a) for 6 h and harvested. B, LNCaP cells were cultured for 12 h in the presence of indicated concentration of DHT or adiol and harvested.
- Fig. 8.
Tumorigenesis of LNCaP cells in severe combined immunodeficient mice. A total of 2 × 106 cells of LNCaP cells were inoculated s.c. in severe combined immunodeficient mice that were castrated and had either castration (cast) alone, testosterone, or androstenediol (adiol) implants. A, tumors were measured from day 20 to day 50 after inoculation. B, after measuring tumor size on day 50 of inoculation, all of the mice were killed; the blood was taken; and the weight of prostate and seminal vesicles (SV) was measured. Data are presented as the mean; bars, ±SE.
Volume 64, Issue 2
