The possibility of aromatization of androgen in human prostate.

Aromatase in human prostate tissue was determined in homogenized human prostate (three BPH and two normal specimens) incubated with [1-beta-3H]androstenedione (radiometric method) or [1,2,6,7-3H]androstenedione (estrogen production analysis method) in the presence of NADPH. Using the former procedure, significant amounts of 3H2O, resulting from the release of 3H at the C-1 position during aromatization, were measured and these increased with incubation time and amount of tissue, whereas the amount of estrone and estradiol-17 beta resulting from the latter method and calculated from the 3H/14C ratio in preparations of purified crystal was very small. The preliminary results, which suggest that an androgen aromatase system exists in the human prostate, point to the need to further investigate the identity and properties of the metabolic products resulting from the conversion of androgen to estrogens and other metabolites.

Effects of busulfan on the DNA of HL-60 cells as a drug sensitivity assay.

We evaluated the formation of DNA-DNA cross links by busulfan as an assay for determining the sensitivity of cells to the drug. HL-60 cells were incubated with busulfan and the effects of the drug on cell growth were compared with the effects of the drug on the alkaline elution pattern of the DNA. While incubation with 100 micrograms/ml for 1 h inhibited cell growth by 50%, drug concentrations of up to 500 micrograms/ml had no effect on the elution pattern of the DNA.

A comparison of estrogen and androgen receptor levels in human prostatic tissue from patients with non-metastatic and metastatic carcinoma and benign prostatic hyperplasia.

Estrogen receptors (ER, N = 72) and androgen receptors (AR, N = 33) were determined by high pressure liquid chromatography (HPLC) in 72 human prostatic tissues obtained at prostatectomy, and exploratory statistical analyses of the resulting data were performed. To facilitate use of these data as well as other pertinent information from the patient charts, a program for a comparatively large data base was implemented on a Wang minicomputer. The median values of cytosolic AR in the four cancer stages examined were statistically different from each other (P = 0.

Correlation of estrogen and androgen receptor status in prostatic disease measured by high pressure liquid chromatography.

To further characterize human prostatic estrogen receptors (ER) determined by high pressure liquid chromatography (HPLC), we checked our procedure by which we nullify estrogen binders (including testosterone binding globulin, TeBG) other than ER by preincubation of cytosols with dihydrotestosterone (DHT). We also showed that the ER exhibited ligand specificity and that ER is present in BPH nuclear extract at 10-fold its concentration in the corresponding cytosols. Of seven prostates with localized cancer determined preoperatively, only 3 showed localization; ER concentration in the cancer parts was lower than in the corresponding surrounding BPH.

A micromethod for the measurement of estrogen receptors using high pressure liquid chromatography.

Adequate sample size has been one of the difficulties encountered in routine determination of estrogen receptors in prostatic disease. The recent commercial availability of protein analysis columns for use with high pressure liquid chromatography (HPLC) equipment has made the achievement of a considerable reduction in sample size feasible. We present a method for determination of ER which includes the use of 16 alpha-125-iodo-estradiol ( [125I]-E2) of 1600 Ci/mmol specific activity, the use of G-25 column chromatography, followed by fractionation by HPLC and finally relation of the fmol specifically bound to mg protein measured in the HPLC peak.

Estrogen receptors and clinical correlations with human prostatic disease.

Measurement of estrogen binding in human prostate using high-pressure liquid chromatography (HPLC) revealed the presence of cytosolic estrogen receptors (ER) both in benign prostatic hyperplasia (BPH) and adenocarcinoma. Receptor concentrations correlated with several histopathologic features in the specimens analyzed. Estrogen receptor levels generally were higher in BPH than in cancer specimens although there was a subgroup of patients with poorly differentiated carcinoma with levels higher than those of BPH, HPLC can be used for measuring ER in 50 microliters of cytosol, and thus needle biopsy specimens will be analyzed routinely for ER with this micromethod.

Estramustine binding in rat, baboon and human prostate measured by high pressure liquid chromatography.

High pressure liquid chromatography (HPLC) was used to determine 3H-estramustine (estradiol-17 beta 3N-bis-[2-chlorethyl] carbamate), 3H-17 beta-hydroxy-5 alpha-androstan-3-one (3H-dihydrotestosterone or 3H-DHT), 3H-estradiol-17 beta (3H-E2) and 3H-3 beta-hydroxy-5-pregnen-20-one (3H-pregnenolone) binding in 50(2) microliter of cytosol utilizing a column which separates proteins in the molecular weight range of 2,000 to 70,000 daltons. The rat prostate contains a protein in considerable concentration and with the highest affinity for estramustine (375,000 dpm 3H-estramustine per mg. cytosol protein) among the substances tested.

Effect of LS 1727, a nitrosocarbamate of 19-nortestosterone, on the R-3327 rat prostatic adenocarcinoma.

LS 1727, a nitrosocarbamate of 19-nortestosterone did not affect the growth of the androgen-dependent R-3327 rat prostate adenocarcinoma. Such treatment markedly increased the weight of the ventral prostate and reduced body weight. The androgenic character of LS 1727 was demonstrated in an experiment in which LS 1727 was found to reduce the uptake of tritiated dihydrotestosterone in both the ventral prostate and the tumors.

Use of silastic implants to raise plasma androgen levels in the immature baboon (Papio cynocephalus).

Two immature Papio cynocephalus baboons were implanted with silastic tubing containing testosterone (T) to obtain prolonged elevation of plasma androgen levels. Plasma radioimmunoassay indicated a thousandfold increase of T levels over baseline for three weeks. Two weeks after implant removal, plasma androgen levels were at, or below, the pre-implantation levels.

Effects of diethylstilbestrol and estramustine phosphate on serum sex hormone binding globulin and testosterone levels in prostate cancer patients.

Serum testosterone-estradiol binding globulin and total testosterone were measured in 2 groups of male controls (less than 50 and more than 65 years old) and in 7 groups of prostatic cancer patients treated with various endocrine manipulation procedures, including orchiectomy, and estramustine phosphate and diethylstibestrol therapy. There were 133 individuals studied. Total serum testosterone levels were significantly higher in the younger versus the older control group and testosterone-estradiol binding globulin levels were significantly higher in the older men.

Measurements of prolactin and androgens in patients with prostatic diseases.

Testosterone (T), dihydrostestosterone (DHT) and prolactin (HPr) levels were determined in normal males and females, in patients with benign prostatic hypertrophy (BPH) and in clinically stable patients with prostatic carcinoma (CAP), intact and orchiectomized. CAP patients were either untreated or on different modalities of therapy. The HPr levels were higher in prostatic cancer patients, in BPH patients, and in subjects on estrogen therapy.

Steroid hormone receptors in the prostate.

Specific receptors for dihydrotestosterone and estradiol-17-beta have been identified in cytosols of the human and baboon prostate. Binding of radioactive estradiol-17-beta to the 0.4 M potassium chloride extractable component of human prostate nuclei also was demonstrated.

A rat prostatic adenocarcinoma as a model for the human disease.

A transplantable, metastasizing prostatic adenocarcinoma (Tumor I) in Lobund Wistar rats was examined for activity and distribution of five hydrolytic enzymes and for ability to accumulate radioactive zinc. The results suggest that the tumor had arisen in the ventral lobe of the prostate and that its growth was not affected by orchiectomy, adrenalectomy, or replacement treatment with exogenous androgen or corticosteroids. The androgen independency of the tumor was further shown by the low uptake of 3H-testosterone, in contrast to the high uptake in the ventral prostate.

Androgen binding in the baboon prostate.

Androgen receptors were identified and partly characterized in cytosols of the caudal and cranial prostatic lobes of a 24-hr castrate baboon. Binding of cyproterone acetate (CA) to testosterone-binding globulin (TeBG) in baboon serum was negligible and therefore was an appropriate unlabeled competitor for distinguishing high-affinity binding of tritiated dihydrotestosterone (3H-DHT) to serum contaminants and receptors in cytosol preparations when multiple-point saturation analyses and removal of free steroid by charcoal adsorption were used. Specificity of androgen binding was demonstrated by the inability of diethylstilbestrol, a synthetic estrogen known to have low binding affinity for TeBG, to displace 3H-DHT from the receptor protein.

Androgen metabolism in sheep.

3H-Testosterone (3H-T) plus 14C-androst-4-ene-3,17-dione (A-dione) and 3H-epi-testosterone (17alpha-hydroxy-4-androsten-3-one) (epiT) plus 14C-T were injected intravenously into two male sheep with bile fistulae, respectively. Urine and bile samples were collected at intervals for 4-8 hours and analyzed by the use of DEAE-Sephadex A-25 and Lipidex 5000 columns, TLC, and paper chromatography; the aglycones were identified by co-crystallization with authentic standards. Five fractions were obtained from urine and bile: unconjugated, glucosiduronates, sulfates, sulfo-glucosiduronates and disulfates.

Androgen metabolism in the rhesus monkey.

A mixture of 3H-testosteron (T) and 14C-4-androstene-3, 17-dione (A) was injected intravenously into 2 (I and II) rhesus monkeys (Macaca mulatta). A third monkey (III) was injected with 3H-T only. Urine and bile samples were collected at intervals for 6 hours following the injection.