Sudden cardiac death (SCD) is associated with both electrical and ischemic substrates, and is a major cause of ischemic heart disease mortality worldwide. Male sex predisposes to SCD but the underlying mechanisms are incompletely understood. KCNE4, a cardiac arrhythmia-associated potassium channel β-subunit, is upregulated by <em>5α</em>-dihydrotestosterone (<em>DHT</em>). Thus, ventricular Kcne4 expression is low in young adult female mice, but high in males and postmenopausal (12+ months) females. Despite causing a sex-independent electrical substrate at 13 months of age (22% QT prolongation in both males and females; P < 0.01), Kcne4 deletion preferentially predisposed aged male mice to ischemia/reperfusion (IR)-provoked ventricular tachyarrhythmias. Interestingly, Kcne4 deletion caused baseline induction of cardioprotective RISK and SAFE pathways in 13-m-old female, but not male, mice. IR-invoked RISK/SAFE induction was also deficient in male but not female Kcne4-/- mice. Pharmacological inhibition of RISK/SAFE pathways in Kcne4-/- females eliminated sex-specific differences in IR-invoked tachyarrhythmia predisposition. Furthermore, castration of Kcne4-/- males eliminated sex-specific differences in both baseline and post-IR RISK/SAFE pathway induction, and tachyarrhythmia predisposition. Our results demonstrate for the first time that male sex can predispose in aged mice to dangerous ventricular tachyarrhythmias despite sex-independent electrical and ischemic substrates, because of testosterone-dependent impairment of RISK/SAFE pathway induction.

In this study, we investigated in an androgenized rat model the involvement of autophagy and mitochondrial dynamics in granulosa cells in the pathogenesis of polycystic ovarian syndrome (PCOS) and its modulation by exogenous gonadotropin (eCG). We found <em>5α</em>-dihydrotestosterone (<em>DHT</em>) treatment reduces ovarian length and weight with predominantly late antral and/or preovulatory stage follicles and no corpora lutea. <em>DHT</em> increased the population of large lysosomes (>50 micron) and macroautophagy, an event associated with granulosa cell apoptosis. Increased granulosa cell Dynamin Related Protein 1 (Drp1) content in the <em>DHT</em> group was accompanied by increased circular and constricted, but reduced rod-shaped, mitochondria. eCG eliminated all atypical follicles and increased the number of late antral and preovulatory follicles with less granulosa cell apoptosis. eCG-treated rats had a higher proportion of connected mitochondria, and in combination with <em>DHT</em> had a lower proportion of circular and constricted mitochondria than rats treated with <em>DHT</em> alone, suggesting that eCG induces mitochondrial fusion and attenuates fission in granulosa cells. In summary, we observed that <em>DHT</em>-induced up-regulation of Drp1 is associated with excessive mitochondrial fission, macroautophagy and apoptosis in granulosa cells at the antral stage of development in an androgenized rat model for PCOS, a response partially attenuated by exogenous gonadotropin.

Alveolar fluid clearance mediates perinatal lung transition to air breathing in newborn infants, which is accomplished by epithelial Na+ channels (ENaC) and Na-K-ATPase. Male sex represents a major risk factor for developing respiratory distress, especially in preterm infants. We previously showed that male sex is associated with reduced epithelial Na+ transport, possibly contributing to the sexual dimorphism in newborn respiratory distress. This study aimed to determine sex-specific effects of sex steroids on epithelial Na+ transport. The effects of testosterone, <em>5α</em>-dihydrotestosterone (<em>DHT</em>), estradiol, and progesterone on Na+ transport and Na+ channel expression were determined in fetal distal lung epithelial (FDLE) cells of male and female rat fetuses by Ussing chamber and mRNA expression analyses. <em>DHT</em> showed a minor effect only in male FDLE cells by decreasing epithelial Na+ transport. However, flutamide, an androgen receptor antagonist, did not abolish the gender imbalance, and testosterone lacked any effect on Na+ transport in male and female FDLE cells. In contrast, estradiol and progesterone increased Na+ transport and Na+ channel expression especially in females, and prevented the inhibiting effect of <em>DHT</em> in males. Estrogen receptor inhibition decreased Na+ channel expression and eliminated the sex differences. In conclusion, female sex steroids stimulate Na+ transport especially in females and prevent the inhibitory effect of <em>DHT</em> in males. The ineffectiveness of testosterone suggests that Na+ transport is largely unaffected by androgens. Thus, the higher responsiveness of female cells to female sex steroids explains the higher Na+ transport activity, possibly leading to a functional advantage in females.

<em>5α</em>-Dihydrotestosterone (<em>5α</em>-<em>DHT</em>) possesses a great affinity for the androgen receptor (AR), and its binding to AR promotes the proliferation of prostate cancer (PC) cells in androgen-dependent PC. Primarily synthesized from testosterone (T) in testis, <em>5α</em>-<em>DHT</em> could also be produced from <em>5α</em>-androstane-3α,17β-diol (3α-diol), an almost inactive androgen, following non-classical pathways. We reported the chemical synthesis of non-commercially available [4-¹⁴C]-3α-diol from [4-¹⁴C]-T, and the development of a biological assay to identify inhibitors of the <em>5α</em>-<em>DHT</em> formation from radiolabeled 3α-diol in LAPC-4 cell PC model. We measured the inhibitory potency of <em>5α</em>-androstane derivatives against the formation of <em>5α</em>-<em>DHT</em>, and inhibition curves were obtained for the most potent compounds (IC₅₀ = 1.2-14.1 μM). The most potent inhibitor 25 (IC₅₀ = 1.2 μM) possesses a 4-(4-CF₃-3-CH₃O-benzyl)piperazinyl methyl side chain at C3β and 17β-OH/17α-CCH functionalities at C17 of a <em>5α</em>-androstane core.

BACKGROUND

Research in the past half a century has gradually sketched the biological mechanism leading to androgenetic alopecia (AGA). Until recently the aetiological paradigm has been too limited to enable intelligent commentary on the use of folk remedies to treat or reduce the expression of this condition. However, our understanding is now at a point where we can describe how some folk remedies work, predict how effective they will be or why they fail.

RESULTS

The new paradigm of AGA is that inheritance and androgens (dihydrotestosterone) are the primary contributors and a secondary pathology, microinflammation, reinforces the process at more advanced stages of follicular miniaturisation. The main protagonist to microinflammation is believed to be microbial or Demodex over-colonisation of the infundibulum of the pilosebaceous unit, which can be ameliorated by antimicrobial/acaricidal or anti-inflammatory therapies that are used as adjuvants to androgen dependent treatments (either synthetic or natural). Furthermore, studies reveal that suboptimal androgen metabolism occurs in both AGA and insulin resistance (low SHBG or high <em>DHT</em>), suggesting comorbidity. Both can be ameliorated by dietary phytochemicals, such as specific classes of phenols (isoflavones, phenolic methoxy abietanes, hydroxylated anthraquinones) or polycyclic triterpenes (sterols, lupanes), by dual inhibition of key enzymes in AGA (<em>5α</em>-reductase) and insulin resistance (ie., DPP-4 or PTP1B) or agonism of nuclear receptors (PPARγ). Evidence strongly indicates that some plant-based folk remedies can ameliorate both primary and secondary aetiological factors in AGA and improve insulin resistance, or act merely as successful adjuvants to mainstream androgen dependent therapies.

CONCLUSIONS

Thus, if AGA is viewed as an outcome of primary and secondary factors, then it is better that a 'multimodal' or 'umbrella' approach, to achieve cessation and/or reversal, is put into practice, using complementation of chemical species (isoflavones, anthraquinones, procyanidins, triterpenes, saponins and hydrogen sulphide prodrugs), thereby targeting multiple 'factors'.

Androgens are a recognized class of endocrine disrupting compounds with the ability to impact reproductive status in aquatic organisms. The current study utilized in vitro exposure of mummichog (Fundulus heteroclitus) testis tissue to either the aromatizable androgen 17α-methyltestosterone (MT) or the non-aromatizable androgen <em>5α</em>-dihydrotestosterone (<em>DHT</em>) over the course of 24 h to determine if there were differential effects on steroidogenic gene expression. Testis tissue was exposed to androgen concentrations of 10-12 M, 10-9 M and 10-6 M for 6, 12, 18 or 24 h, after which a suite of steroidogenic genes, including steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase (3βhsd) and cytochrome P450 17A1 (cyp17a1), were quantified using real-time polymerase chain reaction. Both androgens affected steroidogenic gene expression, with most alterations occurring at the 24-hour time point. The gene with the highest fold-change, and shortest interval to expression alteration, was 3βhsd for both androgens. Potential differences between the two model androgens were observed in increased expression of cyp17a1 and 11β-hydroxysteroid dehydrogenase (11βhsd), which were only altered after exposure to <em>DHT</em> and in expression levels of cytochrome P450 11A1 (cyp11a1), which was upregulated by MT but not altered by <em>DHT</em>. Results from this study show both androgens interact at the gonadal level of the hypothalamus-pituitary-gonadal axis and may possess some distinct gene expression impacts. These data strengthen the current research initiatives of establishing in vitro test systems that allow toxic potential of untested chemicals to be predicted from molecular perturbations.

Androgens originating from pulp mill processing, sewage treatment facilities and agricultural activities have the potential for discharge into aquatic receiving environments. To assess androgen effects on reproductive endocrine status in fish in estuarine environments, male and female adult northern mummichog (Fundulus heteroclitus macrolepidotus) were exposed to an aromatizable androgen (17α-methyltestosterone; MT) and a non-aromatizable androgen (<em>5α</em>-dihydrotestosterone; <em>DHT</em>) in a short-term reproductive endocrine bioassay. Fish were nominally exposed to 10 μg/L or 100 μg/L <em>DHT</em>, or 0.1 μg/L or 1 μg/L MT for 14 days during gonadal recrudescence. Actual concentrations of androgens, as measured by enzyme immunoassay (EIA), were 10-49% of nominal MT 0.1, 3-6% of nominal MT 1, 5-10% of nominal <em>DHT</em> 10 and 3-25% of nominal <em>DHT</em> 100. Female mummichog were impacted to a greater degree by androgen exposure, with increased plasma testosterone (T) at 100 μg/L <em>DHT</em>, depressed plasma 17β-estradiol (E2) at both <em>DHT</em> concentrations and at 1 μg/L MT, as well as depressed in vitro E2 at both MT concentrations and 100 μg/L <em>DHT</em>. Males had depressed plasma T in the 10 μg/L <em>DHT</em> treatment and depressed in vitro 11-ketotestosterone production for both MT concentrations and 10 μg/L <em>DHT</em>. Ovarian aromatase gene expression was induced in females exposed to 1 μg/L MT. <em>DHT</em> at 100 μg/L increased hepatic vitellogenin-1 (VTG1) expression in males and depressed VTG1 expression in females. The range of responses between sexes and among species provides evidence for modes of actions and potential impacts of androgens in aquatic receiving environments.

BACKGROUND

Esophageal adenocarcinoma is a male-dominant disease, but the role of androgens is unclear.

OBJECTIVE

To examine the expression and clinical correlates of the androgen receptor (AR) and the androgen-responsive gene FK506-binding protein 5 (FKBP5) in esophageal adenocarcinoma.

METHODS

Expression of AR and FKBP5 was determined by immunohistochemistry. The effect of the AR ligand <em>5α</em>-dihydrotestosterone (<em>DHT</em>) on the expression of a panel of androgen-responsive genes was measured in AR-positive and AR-negative esophageal adenocarcinoma cell lines. Correlations in expression between androgen-responsive genes were analyzed in an independent cohort of esophageal adenocarcinoma tissues.

RESULTS

There was AR staining in 75 of 77 cases (97 %), and FKBP5 staining in 49 (64 %), all of which had nuclear AR. Nuclear AR with FKBP5 expression was associated with decreased median survival (451 vs. 2800 days) and was an independent prognostic indicator (HR 2.894, 95 % CI 1.396–6.002, p = 0.0043) in multivariable Cox proportional hazards models. DHT induced a significant increase in expression of the androgen-responsive genes FKBP5, HMOX1, FBXO32, VEGFA, WNT5A, and KLK3 only in AR-positive cells in vitro. Significant correlations in expression were observed between these androgen-responsive genes in an independent cohort of esophageal adenocarcinoma tissues.

CONCLUSIONS

Nuclear AR and expression of FKBP5 is associated with decreased survival in esophageal adenocarcinoma.

Ethno pharmacological relevance: Curcuma longa L is traditionally used as an anti-inflammatory remedy in Chinese traditional medicine. Curcuma oil (CO), a lipophilic fraction from Curcuma longa L. has been reported to have anti-proliferative, anti-inflammatory and anti-oxidant activities. However, CO has never been investigated for its possible therapeutic effects on benign prostatic hyperplasia (BPH).

Aims of the study: The study is thus to determine the therapeutic effects of curcuma oil on BPH and also the possible mechanism (s) of action.

<strong class="sub-title"> Materials &methods: </strong> A BPH-1 cell line and Sprague Dawley (SD) rats were used to establish BPH models in vitro and in vivo, respectively. Rats were treated by CO (2.4, 7.2mg/kg/i.g.) and finasteride (5mg/kg/i.g.), respectively. Histological changes were examined by hematoxylin and eosin (H&E) staining. Protein expression was analyzed for <em>5α</em>-reductase (5AR), dihydrotestosterone (<em>DHT</em>), interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α by ELISA. Ki-67, Caspase-8,-9 and -3 expressions were evaluated via immunohistochemistry (IHC).

<strong class="sub-title"> Results: </strong> CO effectively induced apoptosis in BPH-1 cells. BPH was successfully established by administration of testosterone propionate (TP) in rats, which upregulated both <em>5α</em>-reductase expression and <em>DHT</em> production. Importantly, TP establishment significantly stimulated the phosphorylation of p65, one subunit of NF-κB, thus led to activation of the NF-κB signaling pathway in prostatic tissues of rats. In turn, the activation of NF-κB pathway induced concomitant upregulation of proinflammatory factors IL-1β, IL-6, TNF-α, and COX-2 and significant increase of the Bcl2/Bax expression ratio for enhanced cell survival, contributing to the initiation and progression of BPH in rats. Notably, CO therapy significantly decreased prostate weight and hyperplasia in BPH-induced animals. Also CO was found to suppress the expression of <em>5α</em>-reductase and thus the production of <em>DHT</em>, which is essential for the amelioration of BPH. More importantly, CO was shown to suppress the activation of NF-κB pathway through decreasing the expression of phosphorylated p65 and consequently reduced the inflammatory responses and cell survival in prostatic tissues, leading to the inhibition of BPH development in rats.

Conclusion: Curcuma oil is very effective for ameliorating BPH in rats. The underlying mechanisms involve in reduced inflammatory responses and cell survival through suppression of the NF-κB signaling pathway by CO in prostatic tissues.

Keywords: Apoptosis; Benign prostate hyperplasia; Curcuma oil; Inflammation; NF-κB signaling pathway.

Benign prostatic hyperplasia (BPH) is the most common benign tumor in males. Androgen/androgen receptor (AR) signaling plays a key role in the development of BPH; its alterations cause an imbalance between prostate cell growth and apoptosis. Furthermore, chronic inflammation and oxidative stress, which are common conditions in BPH, contribute to disrupting the homeostasis between cell proliferation and cell death. With this background in mind, we investigated the effect of ultramicronized palmitoylethanolamide (um-PEA), baicalein (Baic) and co-ultramicronized um-PEA/Baic in a fixed ratio of 10:1 in an experimental model of BPH. BPH was induced in rats by daily administration of testosterone propionate (3 mg/kg) for 14 days. Baic (1 mg/kg), um-PEA (9 mg/kg) and um-PEA/Baic (10 mg/kg) were administered orally every day for 14 days. This protocol led to alterations in prostate morphology and increased levels of dihydrotestosterone (<em>DHT</em>) and of androgen receptor and <em>5α</em>-reductase expression. Moreover, testosterone injections induced a significant increase in markers of inflammation, apoptosis and oxidative stress. Our results show that um-PEA/Baic is capable of decreasing prostate weight and <em>DHT</em> production in BPH-induced rats, as well as being able to modulate apoptotic and inflammatory pathways and oxidative stress. These effects were most likely related to the synergy between the anti-inflammatory properties of um-PEA and the antioxidant effects of Baic. These results support the view that um-PEA/Baic should be further studied as a potent candidate for the management of BPH.

Keywords: androgen receptor; baicalein; benign prostatic hyperplasia; inflammation; oxidative stress; palmitoylethanolamide.

BACKGROUND

Pig aldo-keto reductase family 1 member C1 (AKR1C1) belongs to AKR superfamily which catalyzes the NAD(P)H-dependent reduction of various substrates including steroid hormones. Previously we have reported two paralogous pig AKR1C1s, wild-type AKR1C1 (C-type) and C-terminal-truncated AKR1C1 (T-type). Also, the C-terminal region significantly contributes to the NADPH-dependent reductase activity for <em>5α</em>-<em>DHT</em> reduction. Molecular modeling studies combined with kinetic experiments were performed to investigate structural and enzymatic differences between wild-type AKR1C1 C-type and T-type.

RESULTS

The results of the enzyme kinetics revealed that Vmax and kcat values of the T-type were 2.9 and 1.6 folds higher than those of the C-type. Moreover, catalytic efficiency was also 1.9 fold higher in T-type compared to C-type. Since x-ray crystal structures of pig AKR1C1 were not available, three dimensional structures of the both types of the protein were predicted using homology modeling methodology and they were used for molecular dynamics simulations. The structural comparisons between C-type and T-type showed that <em>5α</em>-<em>DHT</em> formed strong hydrogen bonds with catalytic residues such as Tyr55 and His117 in T-type. In particular, C3 ketone group of the substrate was close to Tyr55 and NADPH in T-type.

CONCLUSIONS

Our results showed that <em>5α</em>-<em>DHT</em> binding in T-type was more favorable for catalytic reaction to facilitate hydride transfer from the cofactor, and were consistent with experimental results. We believe that our study provides valuable information to understand important role of C-terminal region that affects enzymatic properties for <em>5α</em>-<em>DHT</em>, and further molecular mechanism for the enzyme kinetics of AKR1C1 proteins.

Steroid hormones and receptors play important roles in female reproduction, and their expression patterns affect follicular growth and development. To examine the expression of dihydrotestosterone (<em>DHT</em>) synthases (<em>5α</em>-reductase (<em>5α</em>-red)1 and <em>5α</em>-red2) and androgen receptor (AR) during follicular development, and the regulation of <em>DHT</em> signaling by follicle-stimulating hormone (FSH) and luteinizing hormone (LH), we have used enzyme-linked immunosorbent assays, quantitative real time-polymerase chain reaction, immunohistochemical staining, and western blotting to examine <em>DHT</em> synthesis in small (≤2 mm), medium (2-5 mm), and large (≥5 mm) sheep follicles. Expression of <em>5α</em>-red1, <em>5α</em>-red2, and AR was observed in ovine ovaries, and with the development of follicles, the expressions of <em>5α</em>-red1 and <em>5α</em>-red2 mRNA and protein increased, but the levels of AR mRNA, protein and <em>DHT</em> level decreased. In addition, granulosa cells were treated with FSH (0.01, 0.1, and 1 international unit (IU)/mL), LH (0.01, 0.1, and 1 IU/mL), and testosterone (T, 10^-7 M) to evaluate the effects of FSH and LH on <em>DHT</em> and estradiol (E2) synthesis and <em>5α</em>-red1, <em>5α</em>-red2, and AR expression. We found that FSH and LH up-regulated <em>5α</em>-red1 and <em>5α</em>-red2 in sheep granulosa cells, but down-regulated the concentration of <em>DHT</em> and expression of AR. Meanwhile, FSH and LH significantly up-regulated the expression of aromatase (P450arom) and secretion of E2. This result indicates that although FSH and LH promote the expression of <em>5α</em>-red1 and <em>5α</em>-red2, T is not transformed into <em>DHT</em>, but E2. This study reveals the reason why <em>DHT</em> concentration is down-regulated in large follicles and lays a foundation for further exploring the synthesis mechanism of <em>DHT</em> during follicular development.

Keywords: androgen receptor; dihydrotestosterone; estradiol; granulosa cell; sheep.

Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography-tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC₅₀ values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC₅₀ of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC₅₀ against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.

Keywords: 5α-reductase inhibitors; adverse outcome pathway; dihydrotestosterone; dutasteride; finasteride; in vitro.

The development of targeted therapies has been a consistent goal for hormone-related diseases treatment. As a result of increased knowledge of the role of androgens in different diseases, anti-androgen treatment is becoming increasingly important in targeted therapy. Androgens play an important role in different disorders, therefore, androgen receptor signalling is a crucial factor in pathological conditions. The androgen receptor is a transcription factor activated by the testosterone metabolite <em>5α</em>-dihydrotestosterone and regulates the expression of genes related to sexual differentiation, growth and survival of prostate cells, and to a certain extent, cancer progression. Herein, we review anti-androgen therapies in cancer and other selected diseases and provide examples where anti-androgen drugs can be used as both main and supportive treatments in the multimodal therapeutic scheme. Even in diseases with low serum levels of testosterone or <em>DHT</em>, anti-androgen therapy plays an important role in new treatments. Therefore, the use of anti-androgens is an appealing strategy in which to overcome resistance to primary treatment by assuring better therapy results. In this review, we take into account both older generation hormonal drugs and the new drug classes. Additionally, we review recent studies that suggest new anti-androgen agents have not entirely replaced some of the old standards.

Natural and synthetic steroid hormones, excreted by humans and farmed animals, have been considered as important sources of environmental endocrine disruptors. A suite of estrogens, androgens and progestogens was measured in the wastewater treatment plant outfall (WWTPO) of Chascomús city (Buenos Aires province, Argentina), and receiving waters located downstream and upstream from the WWTPO, using solid phase extraction and high-performance liquid chromatography mass spectrometry. The following natural hormones were measured: 17β-estradiol (E₂), estrone (E₁), estriol (E₃), testosterone (T), <em>5α</em>-dihydrotestosterone (<em>DHT</em>), progesterone (P), 17-hydroxyprogesterone (17OHP) and the synthetic estrogen 17α-ethinylestradiol (EE₂). Also, in order to complement the analytical method, the estrogenic activity in these surface water samples was evaluated using the in vitro transactivation bioassay that measures the estrogen receptor (ER) activity using mammalian cells. All-natural steroid hormones measured, except 17OHP, were detected in all analyzed water samples. E₃, E₁, EE₂ and <em>DHT</em> were the most abundant and frequently detected. Downstream of the WWTPO, the concentration levels of all compounds decreased reaching low levels at 4500 m from the WWTPO. Upstream, 1500 m from the WWTPO, six out of eight steroid hormones analyzed were detected: <em>DHT</em>, T, P, 17OHP, E₃ and E₂. Moreover, water samples from the WWTPO and 200 m downstream from it showed estrogenic activity exceeding that of the EC₅₀ of the E₂ standard curve. In sum, this work demonstrates the presence of sex steroid hormones and estrogenic activity, as measured by an in vitro assay, in superficial waters of the Pampas region. It also suggests the possibility of an unidentified source upstream of the wastewater outfall.

Keywords: Androgens; Aquatic pollution; Endocrine disruption; Estrogens; Municipal wastewater treatment; Progestogens.

OBJECTIVE

Bladder urothelial carcinoma is increasing in incidence with age and its prognosis could become worse when accompanied with metastasis. Effective treatment of these advanced patients is required and it becomes important to understand its underlying biology of this neoplasm, especially with regard to its biological pathways. A potential proposed pathway is androgen receptor (AR)-mediated intracellular signaling but the details have remained relatively unexplored.

METHODS

The expression of AR, <em>5α</em>-reductase type1 (<em>5α</em>R1) and <em>5α</em>-reductase type2 (<em>5α</em>R2) were examined in the bladder cancer cell line T24 and surgical pathology specimens. We also evaluated the status of androgen related cell proliferation and migration using the potent, non-aromatizable androgen agonist <em>5α</em>-dihydrotestosterone (<em>DHT</em>).

RESULTS

<em>DHT</em> treatment significantly increased AR mRNA expression level, but not those of <em>5α</em>R1 and <em>5α</em>R2 in T24 cells. <em>DHT</em> also suppressed cellular migration with weaker and opposite effects on cell proliferation. A significant inverse correlation was detected between pT stage and AR, <em>5α</em>R1 and <em>5α</em>R2 immunoreactivity.

CONCLUSIONS

Inverse correlations detected between tumor grade and AR/androgen metabolizing enzyme also suggested that the loss of AR and androgen-producing enzymes could be associated with tumor progression. Effects of DHT on cells also suggest that androgens may regulate cellular behavior.

<AbstractText>Androgens function through DNA and non-DNA binding-dependant signalling of the androgen receptor (AR). How androgens promote erythropoiesis is not fully understood.</AbstractText><p><div><b>DESIGN AND METHODS</b></div>To identify the androgen signalling pathway, we treated male mice lacking the second zinc finger of the DNA binding domain of the AR (AR^∆ZF2 ) with non-aromatizable <em>5α</em>-dihydrotestosterone (<em>5α</em>-<em>DHT</em>), or aromatizable testosterone. To distinguish direct hematopoietic and non-hematopoietic mechanisms we performed bone marrow reconstitution experiments.</p><p><div><b>RESULTS</b></div>In wild-type mice, <em>5α</em>-<em>DHT</em> had greater erythroid activity than testosterone, which can be aromatized to estradiol. The erythroid response in wild-type mice following <em>5α</em>-<em>DHT</em> treatment was associated with increased serum erythropoietin (EPO) and its downstream target erythroferrone, and hepcidin suppression. <em>5α</em>-<em>DHT</em> had no erythroid activity in AR^∆ZF2 mice, proving the importance of DNA binding by the AR. Paradoxically testosterone, but not <em>5α</em>-<em>DHT</em>, suppressed EPO levels in AR^∆ZF2 mice, suggesting testosterone following aromatization may oppose the erythroid-stimulating effects of androgens. Female wild-type mice reconstituted with AR^∆ZF2 bone marrow cells remained responsive to <em>5α</em>-<em>DHT</em>. In contrast, AR^∆ZF2 mice reconstituted with female wild-type bone marrow cells showed no response to <em>5α</em>-<em>DHT</em>.</p><AbstractText>Erythroid promoting effects of androgens are mediated through DNA binding-dependent actions of the AR in non-hematopoietic cells, including stimulating EPO expression.</AbstractText>

Prostatic enlargement might affect up to 30% of men and can cause signs and symptoms in the lower urinary tract in the elderly. Imbalanced estrogen and androgen secretions are important in prostatic physiopathology. Phthalates-environmental endocrine disruptors-affect androgen secretion and disrupt sexual organs, including testes and the prostate, but the underlying mechanisms are unclear. Using European Association of Urology (EAU) guidelines, we recruited from urology clinics in southern Taiwan 207 elderly men diagnosed with benign prostatic hyperplasia (BPH) and prostatic enlargement between 2015 and 2017. We took blood and urine samples from all patients on the same day. We used multivariate linear regression, associations, and potential interactions after we had measured and analyzed oxidative stress (OS) markers, steroidal hormones, and 11 urinary phthalate metabolites, and then we adjusted for confounders. Di(2-ethylhexyl) phthalate (DEHP) metabolite levels, particularly urinary mono-(2-ethylhexyl) phthalate, were positively associated with androgen, estrogen, hormone ratios, inducible nitric oxide synthetase (iNOS), 8-hydroxy-2'-deoxyguanosine (8-OHdG), prostate specific antigen (PSA), and prostate volume (PV) (p < 0.05). PV and PSA were positively associated with androgen, estrogen, hormone ratios and OS markers (p < 0.05). The estimated percentages of exposure to phthalates in prostatic enlargement mediated by androgen, estrogen, and OS markers ranged from 3.5% to 63.1%. Exposure to DEHP promoted the progress of BPH by increasing dihydrotestosterone (<em>DHT</em>), estradiol (E₂), the converted enzymes aromatase and <em>5α</em> reductase, and reactive oxygen species (ROS) (8-OHdG and iNOS) production. Sex hormones and OS might be important hyperplasia-promoters after a patient has been exposed to phthalates, especially to DEHP.

Axillary osmidrosis is a benign disorder that causes functional and emotional problems in Asian patients. Recently, ApoD has been identified as an axillary odorant binding protein. The present study was designed to compare the expression of ApoD in normal and osmidrosis subjects. Compared with the normal subjects, osmidrosis subjects had a higher expression of AR and ApoD in the apocrine samples, both at mRNA and protein level. Further study showed that, consistent with the increased ApoD and AR, phosphorylated JNK1 was higher in apocrine samples from axillary osmidrosis subjects, while with no obvious differences of the total expression of JNK1. In the cultured apocrine epithelial cells from normal subjects, <em>5α</em>-dihydrotestosterone (<em>5α</em>-<em>DHT</em>) increased the expression of ApoD in a dose dependent manner, which can be inhibited by the JNK1 inhibitor. In contrast, in the cultured apocrine epithelial cells from axillary osmidrosis subjects, inhibition of JNK1 significantly reduced the expression of ApoD. Taken together, our study here revealed that increased JNK1 activation in the apocrine cells from axillary osmidrosis contributes to the increased ApoD expression, which in turn involved in the process of axillary osmidrosis.

BACKGROUND

Benign prostatic hyperplasia (BPH) is non-cancerous condition of enlargement of the prostate, a common occurrence in older men. The immature fruits of Poncirus trifoliata (L.) Rafinesque (Rutaceae), Ponciri Fructus are widely used in traditional oriental medicine for the therapy of various diseases. However, little is known about the mechanism underlying the pathogenesis of BPH. In the present study, we investigated the protective effects of a Ponciri Fructus extract (PFE) on the development of BPH in a in a rat model of BPH induced by testosterone propionate (TP).

METHODS

Male Sprague Dawley rats were used as a model of BPH after its induction by daily subcutaneous injections of TP/corn oil, for a period of four weeks. PFE was administrated daily 1 h before TP/corn oil injection by oral gavage at a dose level of 200 mg/kg during the 4 weeks of TP/corn oil injections. All rats were sacrificed at the end of the experiment, we measured the relative prostate weight, the levels of testosterone and dihydrotestosterone (<em>DHT</em>), histological changes, activities of antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase), and expression of proliferating cell nuclear antigen (PCNA). In addition, we also measured the inhibition (%) of <em>5α</em>-reductase in the prostatic tissue.

RESULTS

Our findings indicate that PFE significantly inhibited the development of BPH; decreased the relative prostate weight, the level of testosterone and <em>DHT</em> in serum and prostatic tissue, prostatic hyperplasia, expression of PCNA, and increased the antioxidant enzymes. Moreover, PFE showed a weak inhibitory activity on <em>5α</em>-reductase.

CONCLUSIONS

These results suggest that PFE may be used as a therapeutic agent for BPH via antiproliferative and antioxidant effects.

Charcoal-stripped fetal bovine serum (CS-FBS) is frequently used in studies on hormone-responsive cancers to provide hormone-free cell culture conditions. CS-FBS may influence the growth of cancer cells; however, the underlying mechanisms remain unclear. In this study, we aimed to clarify the effects of CS-FBS on distinct subtypes of breast cancer cells. We found that the crucial oncoprotein c-Myc was significantly inhibited in estrogen receptor alpha (ER-α)-positive breast cancer cells when cultured in CS-FBS-supplemented medium, but it was not suppressed in ER-α-negative cells. The addition of 17β-estradiol (E2) to CS-FBS-supplemented medium rescued the CS-FBS-induced inhibition of c-Myc, while treatment with <em>5α</em>-dihydrotestosterone (<em>DHT</em>) suppressed c-Myc expression. Our data demonstrated that CS-FBS may impede the growth of ER-α-positive breast cancer cells via c-Myc inhibition, and this was possibly due to the removal of estrogen. These results highlighted that the core drivers of c-Myc expression were subtype-specific depending on the distinct cell context and special caution should be exercised when using CS-FBS in studies of hormone-responsive cancer cells.

Although 11-ketotestosterone (11KT) and testosterone (T) are major androgens in both teleosts and humans, their <em>5α</em>-reduced derivatives produced by steroid <em>5α</em>-reductase (SRD5A/srd5a), i.e., 11-ketodihydrotestosterone (11K<em>DHT</em>) and <em>5α</em>-dihydrotestosterone (<em>DHT</em>), remains poorly characterized, especially in teleosts. In this study, we compared the presence and production of <em>DHT</em> and 11K<em>DHT</em> in Japanese eels and humans. Plasma 11KT concentrations were similar in both male and female eels, whereas T levels were much higher in females. In accordance with the levels of their precursors, 11K<em>DHT</em> levels did not show sexual dimorphism, whereas <em>DHT</em> levels were much higher in females. It is noteworthy that plasma <em>DHT</em> levels in female eels were higher than those in men. In addition, plasma 11K<em>DHT</em> was undetectable in both sexes in humans, despite the presence of 11KT. Three srd5a genes (<i>srd5a1</i>, <i>srd5a2a</i> and <i>srd5a2b</i>) were cloned from eel gonads. All three <i>srd5a</i> genes were expressed in the ovary, whereas only both srd5a2 genes were expressed in the testis. Human <i>SRD5A1</i> was expressed in testis, ovary and adrenal, whereas <i>SRD5A2</i> was expressed only in testis. Human SRD5A1, SRD5A2 and both eel srd5a2 isoforms catalyzed the conversion of T and 11KT into <em>DHT</em> and 11K<em>DHT</em>, respectively, whereas only eel srd5a1 converted T into <em>DHT</em>. <em>DHT</em> and 11K<em>DHT</em> activated eel androgen receptor (ar)α-mediated transactivation as similar fashion to T and 11KT. In contrast, human AR and eel arβ were activated by <em>DHT</em> and11K<em>DHT</em> more strongly than T and 11KT. These results indicate that in teleosts, <em>DHT</em> and 11K<em>DHT</em> may be important <em>5α</em>-reduced androgens produced in the gonads. In contrast, <em>DHT</em> is the only major <em>5α</em>-reduced androgens in healthy humans.

<strong class="sub-title"> Keywords: </strong> 11K<em>DHT</em>; <em>5α</em>-reductase; <em>DHT</em>; androgen receptor; testosterone.

In this study, we investigated steroidogenic gene mRNA expression in human vaginas and verified the ability of human vagina smooth muscle cells (hvSMCs) to synthesize androgens from upstream precursor dehydroepiandrosterone (DHEA). As a readout for androgen receptor (AR) activation, we evaluated the mRNA expression of various androgen-dependent markers.hvSMCs were isolated from vagina tissues of women undergoing surgery for benign gynecological diseases. In these cells, we evaluated mRNA expression of several steroidogenic enzymes and sex steroid receptors using real time RT-PCR. Androgen production was quantified with liquid chromatography tandem-mass spectrometry (LC-MS/MS).In vaginal tissues, AR mRNA was significantly less expressed than estrogen receptor α (ERα), whereas in hvSMCs, its mRNA expression was higher than progestin and both estrogen receptors. In hvSMCs and in vaginal tissue, when compared to ovaries, the mRNA expression of pro-androgenic steroidogenic enzymes (HSD3β1/β2, HSD17β3/β5), along with <em>5α</em>-reductase isoforms and sulfo-transferase, resulted as being more abundant. In addition, enzymes involved in androgen inactivation were less expressed than in the ovaries. The LC-MS/MS analysis revealed that, in hvSMCs, short term DHEA supplementation increased Δ4-androstenedione levels in spent medium, while increasing testosterone (T) and dihydrotestosterone (<em>DHT</em>) secretion after longer incubation. Finally, androgenic signaling activation was evaluated through AR-dependent marker mRNA expression, after DHEA and T stimulation. This study confirmed that the human vagina is an androgen-target organ with the ability to synthesize androgens, thus providing support for the use of androgens for local symptoms of genito-urinary syndrome in menopause.

Keywords: DHEA; GSM; androgens; estrogens; menopause; vagina.

Enhancer RNAs (eRNAs), a subclass of noncoding RNA from enhancers, have biological functions in gene expression. However, their potential role in bladder cancer (BCa) remains largely unknown. The present study investigated the functional role of androgen-associated androgen receptor (AR) mediated-eRNA MARC1 (eMARC1) in BCa progression. Cell proliferation, migration, and apoptosis of BCa cell lines (5637 and T24) with different eMARC1 expression levels or treated with <em>5α</em>-dehydrotestosterone (<em>DHT</em>) were investigated. In the current study, we discovered that eMARC1 was highly expressed in BCa tissues and cell lines, and eMARC1 overexpression promoted the progression of BCa cells, while knockdown of eMARC1 suppressed tumorigenesis. <em>DHT</em> treatment significantly elevated eMARC1 expression levels, which also facilitated cell proliferation, motility, and inhibited cell apoptosis. We further found that eMARC1 silencing impaired the androgenic effect of <em>DHT</em> in BCa cells. These results suggested that eMARC1 exerted its effects on BCa cell progression, and <em>DHT</em> promoted bladder cancer progression by activating eMARC1.