The role of non-aromatizable testosterone metabolite in metabolic pathways

Dihydrotestosterone (DHT) originates via irreversible reduction of testosterone by catalytic activity of 5alpha-reductase enzyme and it is demonstratively the most effective androgen. Androgens influence adipose tissue in men either directly by stimulation of the androgen receptor or indirectly, after aromatization, by acting at the estrogen receptor. DHT as a non-aromatizable androgen could be responsible for a male type fat distribution. The theory of non-aromatizable androgens as a potential cause of a male type obesity development has been studied intensively. However, physiological levels of DHT inhibit growth of mature adipocytes. In animal models, substitution of DHT in males after gonadectomy has a positive effect on body composition as a testosterone therapy. Thus, DHT within physiological range positively influences body composition. However, there are pathological conditions with an abundance of DHT, e.g. androgenic alopecia and benign prostatic hyperplasia. These diseases are considered as risk factors for development of metabolic syndrome or atherosclerosis. In obese people, DHT metabolism in adipose tissue is altered. Local abundance of non-aromatizable androgen has a negative effect on adipose tissue and it could be involved in pathogenesis of metabolic and cardiovascular diseases. Increased DHT levels, compared to physiological levels, have negative effect on development of cardiovascular diseases. Difference between the effect of physiological and increased level brings about certain paradox.     

The clinical development of a 5 alpha-reductase inhibitor, finasteride.

Finasteride, a 4-aza steroid compound, is an orally active inhibitor of the 5 alpha-reductase enzyme. 5 alpha-Reductase is necessary for the metabolism of testosterone (T) to dihydrotestosterone (DHT) and is found in high levels only in certain tissues such as the prostate. Finasteride has been shown to markedly suppress serum DHT levels in man without lowering testosterone levels. In patients with benign prostate hyperplasia (BPH), finasteride was found to decrease prostate volume by a mean of 28% over a period of 6 months, without causing clinically significant adverse effects. DHT appears to be the primary androgen for prostatic growth. Selective inhibition of 5 alpha-reductase by finasteride may provide a novel approach to BPH therapy by reducing prostate size without affecting T-dependent processes such as fertility, muscle strength, and libido. The clinical development of finasteride for the treatment of benign prostate hyperplasia is reviewed.     

Ammonium sulfate precipitation as a tool for the study of androgen receptor proteins in rat prostate and mouse kidney.

Ammonium sulfate precipitation has been used for the separation of bound and free steroids in rat prostate and mouse kidney cytosol equilibrated with tritiated androgens. A high affinity, low capacity binding protein has been identified in the 35% saturation precipitate. Biochemical and physiological data indicate that this protein is identical with the previously described 8-10 S androgen receptor. It has been demonstrated that this receptor protein binds 17 beta - hydroxy-5alpha-androstan-3-one (DHT) and testosterone in both tissues. The apparent dissociation constant (Kd) of the prostatic receptor for DHT and of the renal receptor for testosterone is 1-2 nM. The number of binding sites equals 57 and 23 fmoles/mg protein in prostate and kidney respectively. Dterminations of apparent inhibition constants (Ki) for 26 steroidal and non-steroidal compounds suggest that the binding sites in these tissues is similar or identical.     

The presence of a hitherto undefined high-capacity androgen binding macromolecule in human ovarian cancer tissue

Sex hormone binding globulin (SHBG) is known to interfere in the quantitation of androgen receptors (AR) if dihydrotestosterone (DHT) is used. We used a monoclonal antibody to remove SHBG from cytosol. In cytosol of benign prostatic hyperplastic (BPH) tissue low capacity binding for DHT, but not for R1881, was found after removal of SHBG. AR were detected in 18 of 20 ovarian cancer cytosols. In the two AR-negative cases, non-saturable binding for DHT, testosterone and R1881 was observed. Incubation with anti-SHBG did not change this. An hitherto undefined androgen binding macromolecule(s), with high-capacity binding for natural and synthetic androgens, but not for estrogen and progesterone, seems to be present in these ovarian cancer tissues. The functionality of these androgen binding macromolecules in ovarian cancer is yet to be demonstrated.     

Steroidal influences on pup-killing behavior in mice

Four experiments were conducted in order to examine the biochemical pathway through which testosterone (T) acts to produce pup-killing behavior in ovariectomized female mice. Estradiol benzoate (EB) and testosterone propionate (TP) were both effective in stimulating the pup-killing response (Experiment 1). The nonaromatizable androgen dihydrotestosterone (DHT) and the aromatizable androgen testosterone (T) were equipotent in eliciting the behavior while the nonaromatizable androgen androstanedione (AA) was ineffective (Experiment 2). Pup-killing behavior was activated by the aromatizable androgen androstenedione (AE) but only if it was administered chronically at very high doses (Experiment 3). The combination of subthreshold doses of EB and DHT synergized to produce the pup-killing response (Experiment 4). These findings suggest that both aromatization and reduction may be important for the stimulation of pup-killing behavior in mice.     

Pretreatments with 5 alpha-dihydrotestosterone and the uptake of testosterone by cell nuclei in the brains of male rhesus monkeys

An in vivo competition method was used in adult male rhesus monkeys to determine if testosterone binds to high affinity binding agents, notably androgen receptors, in brain cell nuclei. Castrated males received 5 alpha-dihydrotestosterone propionate (DHTP, 20 mg, N = 6), testosterone propionate (TP, 100 mg, N = 3) or oil vehicle (controls, N = 6) followed 3 h later by 5 mCi [3H]testosterone [( 3H]T) as an intravenous bolus. Brain and peripheral tissue samples were removed after 60 min, homogenized and separated into supernatant and purified nuclear fractions. Radioactive metabolites of [3H]T [( 3H]estradiol, [3H]DHT) and unchanged [3H]T were identified by high performance liquid chromatography (HPLC). Androgen pretreatments reduced the nuclear uptake of [3H]T by 67-98% in hypothalamus (HYP), preoptic area (POA) and pituitary gland (PIT). This blockade was presumed to be due to prior occupation of nuclear androgen receptors by unlabeled androgens because pretreatments had no effects on levels of [3H]T in supernatants. Since [3H]T was the major radioactive androgen present in brain cell nuclei, results strongly suggested that the principal nuclear androgen receptor ligand in HYP, POA and PIT was unchanged [3H]T rather than [3H]DHT as occurs in the genital tract. In the amygdala the situation was quite different. Here, nuclear concentrations of [3H]T were reduced by 67% following TP pretreatment but were not changed following DHTP pretreatment, indicating a different uptake mechanism in this region that could have particular relevance for testosterone's central actions on behavior.     

Androgen and estrogen dynamics in the female baboon (Papio anubis)

Androgen and estrogen dynamics were studied in 5 female baboons (Papio anubis) using constant infusions of [3H]androstenedione/[14C]estrone and [3H]testosterone/[14C]estradiol. Blood samples were obtained prior to the infusions and both blood and plasma was used for measurements of androstenedione (A), testosterone (T), dihydrotestosterone (DHT), estrone (E1), estradiol (E2). Plasma was used for measurements of sex-hormone binding globulin (SHBG), and the percents of T and E2 free, bound to SHBG, and to albumin. Blood samples obtained during the infusions were analyzed for radioactivity as purified androgens and estrogens. Metabolic clearance rates (MCR), and transfer factors ([rho]BB; fraction of steroid infused which is converted to and measured in blood as product) and blood production rates were calculated from whole blood data. All urine was collected for 96 h and an aliquot analyzed for radioactivity as the glucuronides of estrone and estradiol and the % peripheral aromatization calculated. The MCR's, calculated in whole blood, of A, E1, E2 and T were 53 +/- 6 1/day/kg, 39.3 +/- 3 1/day/kg, 29.9 +/- 5.2 1/day/kg and 10.1 +/- 2.3 1/day/kg, respectively. Each MCR was different (P less than 0.05) from the others. The PB of E1 was 15 +/- 2 micrograms/day and was not different from that of E2 (12 +/- 3 micrograms/day). The PB of A, 231 +/- 55 micrograms/day, was greater than that of T, 13 +/- 5 micrograms/day. The interconversions of both the androgens (18.9 +/- 3.4% vs 3.9 +/- 1.0%) and the estrogens (48.8 +/- 10.7% vs 4.0 +/- 0.8%) favored the oxidative pathway, i.e. conversion of 17-OH to 17-oxo steroids. The conversion ratio of A to DHT was greater than that of T to DHT (16.4 +/- 2.1% vs 5.3 +/- 0.7%), and A is a more important source of DHT than is T. The percent of T bound to SHBG (80.7 +/- 0.9%) was greater than percent of E2 (36.9 +/- 9.8%) and inversely the percents of T bound to albumin and free (17.5 +/- 0.8% and 1.65 +/- 0.16%) were less than the respective percents for estradiol (60.5 +/- 9.5% and 2.40 +/- 0.27%). The mean SHBG concentration was 54 +/- 6 nM. The peripheral aromatization of androstenedione, 1.36 +/- 0.05%, was greater than of testosterone, 0.18 +/- 0.02%. This difference is, in part, due to the lack of SHBG-binding of androstenedione. The general pattern of androgen and estrogen dynamics is similar to that in women. This similarity is due, in part, to the presence of SHBG in both baboons and women.     

Plasma steroids in benign prostatic hypertrophy and carcinoma of the prostate

Cortisol, 17-hydroxyprogesterone (17-OH-P), progesterone, testosterone, 5 alpha-dihydrotestosterone (DHT), estrone (E1) and estradiol (E2) were determined by RIA after chromatographic separation of steroids on Sephadex LH-20 columns, in 54 hospitalized patients with benign prostatic hyperplasia (BPH) and in 32 hospitalized patients with prostatic carcinoma (PCA) (T34, N01, M01). The patients' values were compared with those of 63 age-matched controls. Increased cortisol and DHT levels, subnormal estrogen and 17-OH-P values and normal progesterone level were found in both benign and malignant groups. Higher mean values for testosterone, and T/DHT ratio and lower mean values for E2/T ratio were found in PCA, as compared with those in BPH. An age invariance of cortisol, testosterone, T/DHT ratio and estradiol was found in both BPH and PCA, instead of the age dependence found in normal subjects. The normal relations between testosterone and its precursor (17-OH-P) and its peripheral metabolite (E2), respectively, and the normal relation between estrone and 17-OH-P were not evident in either BPH or PCA group. The normal direct relation between testosterone and DHT has been found in both patient groups.     

Bone tumors and androgen metabolism]

The paper provides data of comparative assessment of activity of the key enzymes in metabolism of androgens in various morphological variants of sarcomas and benign bone tumors in 46 patients within the age range from 15 to 61, which were under treatment in the ORC named after N. N. Blokhin, RAMS, from 1996 to 1997. On the basis on publications and our own research results we can suggest that malignant human bone tumors of various histogenesis are subject to metabolic processes of testosterone (T), the principal androgen regulator in the bone tissue, as well as formation in bones of 5 alpha-dihydrotestosterone (DHT). One cannot exclude a possibility that metabolism of androgen in bone tissues is directed towards formation of other androgens (in addition to DHT), which may participate in the bone tissue regulation, for instance, 3 alpha- and 3 beta-diols; specific activity of the latter has recently been intensively scrutinized. One can expect that further research shall disclose the clinical significance of metabolism of androgens and of individual androgens in human bone formations.     

Testosterone and dihydrotestosterone concentrations in elephant serum and temporal gland secretions

Serum and temporal gland secretions (TGS) were obtained from mature wild African (Loxodonta africana) and captive Asian elephants (Elephas maximus). Samples were obtained from five cows and eight bulls culled for management purposes in Kruger National Park, South Africa, and from four females and two males residing at the Washington Park Zoo, Portland, Oregon. Our purpose was to describe the levels of the androgens, testosterone (T), and dihydrotestosterone (DHT), and to correlate these observations with sex, species and behavioral status. Male-female differences in serum T were pronounced in the Asian species, whereas male and female concentrations overlapped in the African elephant serum. Serum T concentrations in African females were greater than in Asian females. Serum DHT reflected T levels, except that the striking elevation of testosterone in Asian bulls during musth was not paralleled by equal increases in DHT levels. A species difference observed among males was higher serum T levels in nonmusth Asian bulls (1.84-5.35 ng/ml) compared to the levels in African bulls (0.38-0.68 ng/ml), except for one dominant African bull (6.64 ng/ml). This single African value was still considerably lower than the serum T values of the Asian males during musth. These musth values were the highest serum androgen concentrations: T was between 19 and 40 ng/ml (average 26.10 ng/ml). The TSG values of T and DHT were much higher than serum levels except in the Asian female. T/DHT ratios in TGS were more similar than in serum. One dominant African bull had a T TGS value of 78 ng/ml, which was much higher than the rest of the African males or females, but considerably lower than as Asian bull in musth (547 ng/ml). It seems apparent that a change in androgen status as reflected in serum and TGS levels of T and DHT precedes or is concomitant with overt alteration in behavior in the Asian male. The temporal gland appears to actively concentrate androgens in both African males and females, but in the Asian male the gland secretes only during musth when the greatest concentration of both T and DHT were observed. The apparent difference in the degree of temporal gland secretory activity between the two species suggests a more specific communicative function within the Asian male.     

Comparison of the nuclear 5 alpha-reduction of testosterone and androstenedione in human prostatic carcinoma and benign prostatic hyperplasia.

The nuclear conversion of testosterone (T) to dihydrotestosterone (DHT) and androstenedione (delta 4A) to androstanedione (5 alpha-Adione) was compared in the separated stromal and epithelial fractions of hyperplastic (n = 6) and malignant (n = 3) prostatic tissues. Assay conditions were linear with respect to time and protein concentration and were optimal for NADPH concentration. The apparent Km values for the stromal enzymes were 0.2 and 0.02 microM for hyperplasia and carcinoma, respectively, using T as substrate. The apparent Km values, using delta 4A as substrate, were 0.03 and 0.02 microM, respectively. Apparent Vmax values for the stromal formation of DHT were 16.5 +/- 5.4 and 1.97 +/- 0.45 pmol/mg protein/30 min incubation, respectively, for the hyperplastic and malignant tissues. The apparent Vmax values for the formation of 5 alpha-Adione were 2.8 +/- 1.3 and 6.5 +/- 1.2 pmol/mg/protein/30 min incubation. The apparent Km values for the epithelial enzyme, for hyperplastic and malignant tissue were 0.04 and 0.04 microM, for T, and 0.05 and 0.03 microM for delta 4A. The respective apparent Vmax values were 4.6 +/- 0.93 and 0.65 +/- 0.07 for DHT and 2.0 +/- 0.86 and 6.4 +/- 0.45 pmol/mg protein/30 min incubation for 5 alpha-Adione. delta 4A was a competitive inhibitor of T 5 alpha-reduction. These results provide further evidence that different rates of 5 alpha-reduction at least partially explain the differences in androgen levels seen in the hyperplastic and the malignant prostate.     

Androgen- and estrogen-induced copulatory behavior and inhibition of luteinizing hormone (LH) secretion in the male rat

The purpose of the present investigation was to determine if estrogen, aromatizable androgen or nonaromatizable androgen is capable of (1) inducing copulatory behavior and (2) inhibiting the postcastration rise in plasma LH. Castrate male rats were injected daily with either 1 mg testosterone (T), androstenedione (A), dihydrotestosterone (DHT), or 25 μg estradiol benzoate (EB) or oil and tested weekly for masculine behavior and for lordosis behavior after 38 days of steroid treatment. On day 40 blood was collected for radioimmunoassay of plasma LH. At least 89% of the males treated with T, A, or EB and 55% of those treated with DHT displayed ejaculatory behavior whereas none of the oil-treated males showed male copulatory behavior. Only estrogen-treated males displayed lordosis behavior. T and to a lesser extent A treatment reduced high levels of plasma LH; however, DHT and EB further reduced plasma LH to undectable levels. The relative potency of the steroid effect in stimulating accessory sex tissues followed the order: DHT > T > A > EB = oil. Significant dissociation was observed between the effects of these steroids on peripheral morphology, negative feedback, and mating behavior. These results indicate that masculine behavior is facilitated to the greatest extent, although not exclusively, by centrally acting aromatizable androgen or estrogen, whereas under the present conditions only estrogen stimulates feminine behavior.     

Regional differences in the testosterone to dihydrotestosterone ratio in the epididymis and vas deferens of adult mice

The concentrations of testosterone and dihydrotestosterone (DHT) were measured in the testis and in different segments of the epididymis and vas deferens of adult mice. There were marked regional variations in the concentrations of testosterone and DHT from the testis to the caudal part of the vas deferens. In the testis, testosterone was the predominant androgen (364 +/- 90 ng/g) while DHT was weakly represented (8 +/- 2 ng/g). Qualitative and quantitative changes occurred in epididymis: DHT was the main steroid in the caput (29.3 +/- 2.7 ng/g) and corpus (33.1 +/- 4.4 ng/g) while testosterone and DHT were in similar quantities in the cauda (18.6 +/- 2.6 and 19.0 +/- 2.7 ng/g, respectively). The proximal region of the vas deferens contained higher amounts (71.4 +/- 8.0 ng/g) of androgens (testosterone + DHT) than did the caput epididymidis (39.1 +/- 3.3 ng/g). Testosterone was the predominant androgen in each part of the vas deferens and its concentrations decreased from the proximal (64.5 +/- 7.5 ng/g) to the caudal (26.9 +/- 4.3 ng/g) region. Castration and section of the efferent ducts of the testis showed that the epididymis received testosterone essentially via the blood supply and that epididymal DHT was produced locally from circulating testosterone.     

Nicotine and cotinine effects on 3 alpha hydroxysteroid dehydrogenase in canine prostate

We have recently observed that cigarette smoking affects plasma androgen concentrations. The effects of nicotine and cotinine, two products of cigarette smoking, on testosterone metabolism were determined. The activity of delta 4 steroid 5 alpha-reductase, which converts testosterone to 5 alpha-dihydrotestosterone (DHT) was measured in isolated dog prostate nuclei using testosterone (0-200 nM) as substrate and NADPH as cofactor. Activity of 3 alpha-hydroxysteroid dehydrogenase (HSD), which converts DHT to 3 alpha-androstanediol (3 alpha-diol) and is a reversible enzyme, was measured in isolated dog prostate microsomes with DHT (0-20 microM) as substrate and NADPH as cofactor. When microsomal fractions were incubated for 1 hour with and without nicotine (0-50 microM) and cotinine (0-100 microM), enzyme activity of HSD was significantly suppressed (p less than 0.001). The Vmax was not affected significantly (p greater than 0.60) and Km increased with increasing concentrations of nicotine and cotinine (p less than 0.05). Both nicotine and cotinine are competitive inhibitors of HSD in dog prostate microsomes with Ki's of 61 and 89 microM, respectively. The apparent 5 alpha-reductase activity was unaffected by nicotine and cotinine. The inhibitors produced a marked effect on activity of HSD when used in concentrations achieved in humans who smoke cigarettes. The results suggest that nicotine and cotinine are competitive inhibitors of the HSD, an important enzyme involved in the metabolism of DHT and produce an accumulation of DHT. These products of cigarette smoking could alter androgen action in tissue such as skin and prostate.     

Neural uptake and metabolism of testosterone and dihydrotestosterone in the guinea pig

Neural tissues from adult, castrated male guinea pigs were examined for their capability to concentrate and metabolize [1,2-³H]testosterone (T) and [1,2-³H]dihydrotestosterone (DHT), both in vitro and in vivo. In vitro uptake of DHT and T was greater in the hypothalamus and anterior pituitary than in the cerebral cortex. With DHT as the substrate, the 800×g particulate concentration of this compound was highest in the hypothalamus, although in this tissue, particulate concentration was less than that of the cytoplasm. In the cerebral cortex 5α-androstane-3,17-dione was the most abundant metabolite, whereas 5α-androstane-3,17-dione, 5α-androstane-3α,17β-diol, and 5α-androstane-3β,17β-diol were all present in equivalent amounts in the hypothalamus and pituitary. Incubation with T resulted in the formation of DHT, 4-androstane-3,17-dione, and a compound with the mobility of 5α-(or 5β-)androstane-3,17-7-dione. The radioactivity associated with DHT was the most prevalent in the pituitary (1.3%), and least prevalent in the cerebral cortex (0.6%), and in all cases cytoplasmic concentration of this compound exceeded the concentration in the particulate fraction. Recrystallization failed to confirm the presence of estradiol-17β. Although there were no apparent tissue differences in the uptake of DHT or T 1 hour after their injection, intracellular distribution varied. In all tissues examined, that percentage of total radioactivity attributable to DHT was greatest in the 800×g particulate preparations, particularly in the hypothalamus. Thus neural tissues in the guinea pig, as in other species, exhibit differential uptake and metabolism of androgen through which physiological and behavioral effects may be mediated.     

The hormonal control of begging and early aggressive behavior: experiments in black-headed gull chicks

The hormonal control of begging and sibling competition is largely unknown, but recent evidence suggests a role for steroid hormones. We tested the influence of the aromatizable androgen testosterone (T), the non-aromatizable androgen 5alpha-dihydrotestosterone (DHT), and 17beta-estradiol (E) on both begging behavior and aggressive behavior in black-headed gull chicks (Larus ridibundus). Chicks of this species have a conspicuous begging display, while their frequently performed early aggressive behavior is facilitated by testosterone and important for territorial defense. Hormone treatment was applied by implants between days 6 and 16 after hatching. Behavior was tested by means of standard stimulus tests. The results were validated in a second experiment under semi-natural conditions. Begging was suppressed by T and DHT and not affected by E. Aggressive Pecking was strongly facilitated by T. The erect threat posture, characteristic for older chicks, was facilitated by T, DHT, and E and the nest-oriented threat display, typical for young chicks, only by T and DHT. Growth was suppressed in the T group. The results indicate that androgen production, needed for territorial defense, has costs in terms of a suppression of begging and growth. It is discussed to what extent older chicks may avoid these costs by converting testosterone to estrogen and why pre-natal and post-natal exposure to androgens differ in their effect on begging behavior.     

Differential regulation of gonadotropin-releasing hormone secretion and gene expression by androgen: membrane versus nuclear receptor activation

Steroid hormones induce rapid membrane receptor-mediated effects that appear to be separate from long-term genomic events. The membrane receptor-mediated effects of androgens on GT1-7 GnRH-secreting neurons were examined. We observed androgen binding activity with a cell-impermeable BSA-conjugated testosterone [testosterone 3-(O-carboxymethyl)oxime (T-3-BSA)] and were able to detect a 110-kDa protein recognized by the androgen receptor (AR) monoclonal MA1-150 antibody in the plasma membrane fraction of the GT1-7 cells by Western analysis. Further, a transfected green fluorescent protein-tagged AR translocates and colocalizes to the plasma membrane of the GT1-7 neuron. Treatment with 10 nM 5alpha-dihydrotestosterone (DHT) inhibits forskolin-stimulated accumulation of cAMP, through a pertussis toxin-sensitive G protein, but has no effect on basal cAMP levels. The inhibition of forskolin-stimulated cAMP accumulation by DHT was blocked by hydroxyflutamide, a specific inhibitor of the nuclear AR. DHT, testosterone (T), and T-3-BSA, all caused significant elevations in intracellular calcium concentrations ([Ca(2+)](i)). T-3-BSA stimulates GnRH secretion 2-fold in the GT1-7 neuron, as did DHT or T. Interestingly GnRH mRNA levels were down-regulated by DHT and T as has been reported, but not by treatment with T-3-BSA or testosterone 17beta-hemisuccinate BSA. These studies indicate that androgen can differentially regulate GnRH secretion and gene expression through specific membrane-mediated or nuclear mechanisms.     

New approaches for the treatment of prostatic hypertrophy and cancer

The present study reports the effects exerted by 4-hydroxy-4-androstene-3,17-dione (4-OH-A) on the in vitro metabolism of labelled testosterone, dihydrotestosterone (DHT) and androstenedione (delta-4-A) in the prostate of adult male rats and in human benign prostatic hypertrophic (BPH) tissue. It has been found that 4-OH-A decreases the formation of DHT and of the diols. When testosterone is used as the substrate, the presence in the medium of 4-OH-A enhances the formation of delta-4-A and of 5-alpha-androstanedione (5-alpha-A); 4-OH-A does not inhibit the conversion of labelled DHT into the diols. Also, the transformation of labelled delta-4-A into 5-alpha-A is not modified by 4-OH-A. On the basis of these findings, it is suggested that 4-OH-A might represent a potential new agent for the prevention and/or treatment of human BPH.     

Effect of castration monotherapy on the levels of adrenal androgens in cancerous prostatic tissues

The mechanism accounting for the development of castration-resistant prostate cancer (CRPC) remains unclear. Studies in CRPC tissues suggest that, after androgen deprivation therapy (ADT), the adrenal androgens may be an important source of testosterone (T) and 5-alpha dihydrotestosterone (DHT) in CRPC tissues. To clarify the role of adrenal androgens in the prostatic tissues (prostatic tissue adrenal androgens) during ADT, we developed a high sensitive and specific quantification method for the levels of androgens in prostatic tissue using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Human prostatic tissues were purified using mixed-mode reversed-phase, strong anion exchange Oasis cartridges (Oasis MAX). Analysis of steroids was performed using LC-MS/MS after picolinic acid derivatization. The validation tests showed that our method of quantitative analysis was precise and sensitive enough for the quantification of dehydroepiandrosterone (DHEA), androstenedione, androstenediol, T, and DHT in the prostatic tissue. The levels of adrenal androgens in prostate cancer tissues after ADT were similar to those in untreated PCa. Especially, DHEA was the most existing androgen precursor in PCa tissues after ADT. The levels of DHEA were high in PCa tissues, irrespective of ADT. We assumed that DHEA played a significant role in the synthesis of T and DHT in PCa tissues after ADT.