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1.
Anticancer Res ; 40(5): 2559-2565, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32366400

RESUMO

BACKGROUND/AIM: Androgens are essential for the growth of most prostate cancers (PCa). As a result, androgen ablation is the mainstay of the treatment of PCa. Proteins of the polycomb and trithorax family are master epigenetic regulators of cell type specific gene expression including androgen receptor. MATERIALS AND METHODS: We interrogated epigenetic changes of a 24-gene panel corresponding to polycomb and trithorax genes by PCR array and differential gene expression by quantitative real time-PCR on prostate cancer cell line (LNCaP) treated with the synthetic ligand R1881. RESULTS: We observed the highest methylation for CBX2, PCGF6, PHC2, EZH2 and TRIM27 genes and the lowest methylation for CBX8 and PCGF2 (p<0.05), and a modest decrease in the expression of EZH2. CONCLUSION: Differential methylation profiles of polycomb and trithorax genes may contribute to the dynamics of prostate carcinogenesis.


Assuntos
Androgênios/farmacologia , Epigênese Genética/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Histona-Lisina N-Metiltransferase/genética , Proteína de Leucina Linfoide-Mieloide/genética , Proteínas do Grupo Polycomb/genética , Neoplasias da Próstata/genética , Androgênios/metabolismo , Linhagem Celular Tumoral , Metilação de DNA , Humanos , Masculino , Receptores Androgênicos/metabolismo
2.
Endocrinology ; 161(1)2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31875912

RESUMO

Changes in gonadotropin-releasing hormone (GnRH) release frequency from the brain help drive reproductive cycles. In polycystic ovary syndrome (PCOS), persistent high GnRH/luteinizing hormone (LH) frequency disrupts cycles and exacerbates hyperandrogenemia. Adult prenatally-androgenized (PNA) mice exhibit increased GnRH neuron firing rate, elevated ovarian androgens, and disrupted cycles, but before puberty, GnRH neuron activity is reduced in PNA mice compared with controls. We hypothesized that ovarian feedback mediates the age-dependent change in GnRH neuron firing rate in PNA vs control mice. Extracellular recordings of green fluorescent protein (GFP)-identified GnRH neurons were made 5 to 7 days after sham-surgery, ovariectomy (OVX), or, in adults, after OVX plus replacement of sub-male androgen levels with dihydrotestosterone implants (OVX + DHT). In 3-week-old mice, OVX did not affect GnRH neuron firing rate in either group. In adult controls, OVX increased GnRH neuron firing rate, which was further enhanced by DHT. In adult PNA mice, however, OVX decreased GnRH neuron firing rate, and DHT restored firing rate to sham-operated levels. In contrast to the differential effects of ovarian feedback on GnRH neuron firing rate, serum LH increased after OVX in both control and PNA mice and was not altered by DHT. Pituitary gene expression largely reflected changes expected with OVX, although in PNA but not control mice, DHT treatment increased Lhb expression. These results suggest prenatal androgen exposure programs marked changes in GnRH neuron regulation by homeostatic steroid feedback. PNA lowers GnRH neuron activity in low-steroid states (before puberty, OVX), and renders activity in adulthood dependent upon ongoing exposure to elevated ovarian androgens.


Assuntos
Androgênios/farmacologia , Hormônio Liberador de Gonadotropina/metabolismo , Neurônios/fisiologia , Ovário/metabolismo , Animais , Di-Hidrotestosterona/farmacologia , Eletrofisiologia , Feminino , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Ovariectomia , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Maturidade Sexual/fisiologia
3.
Meat Sci ; 159: 107916, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31476680

RESUMO

Testosterone, as an influential factor in marbling score, requires strict management for uniform development of adipocytes in-between muscle bundles. Present study investigated effect of castration timing and testosterone levels on adipocyte development using SVCs. Isolated SVCs exhibited classical MSC markers, CD31-, CD34-, CD45-, CD90+, and CD105+. ELISA analysis indicated that serum testosterone concentration was highest in non-castrated calves while no significant difference was observed between female, early and late castrated calves. CCK-8 assay showed that concentration of testosterone had no effect on cell proliferation. However, the real-time PCR demonstrated that 20 ng/ml of testosterone suppressed expression of preadipocyte markers, pref-1 and zfp423, but encouraged expression of myoblast markers, myf5 and myoD, via the AR. Consequently, expression of adipogenic markers C/EBPα and PPARγ, as well as accumulation of triglyceride, were decreased in 20 ng/ml testosterone treatment under adipogenic conditions. These findings suggest that by castrating calves before level of testosterone increases, may improve marbling development in the Hanwoo beef industry.


Assuntos
Bovinos , Músculo Esquelético/citologia , Testosterona/farmacologia , Adipócitos , Androgênios/administração & dosagem , Androgênios/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Masculino , Orquiectomia/veterinária , Testosterona/administração & dosagem
4.
J Urol ; 203(5): 940-948, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31845837

RESUMO

PURPOSE: Beyond testosterone, several steroids contribute to the activation of the androgen receptor pathway, but their relative contributions to the activation of the androgen receptor signaling axis in patients with castrated prostate cancer remain unknown. MATERIALS AND METHODS: Serum levels of 9 steroids were measured by mass spectrometry from continuously castrated patients of the PR.7 study (219) and from the PCA24 cohort (116). For each steroid standard curves for dose dependent prostate specific antigen promoter activation were built in castration sensitive (LAPC4) and resistant (VCaP) prostate cancer models. Standard curves were used to determine the androgen receptor activation potency for each steroid measurement from patients in these trials. RESULTS: In LAPC4 and VCaP cells testosterone, dihydrotestosterone and androstenedione induced androgen receptor transcriptional activity, while dehydroepiandrosterone, 5alpha-androstan-3beta,17beta-diol, androstenediol and androsterone stimulated androgen receptor only in VCaP cells. Extragonadal steroids were responsible for 34% (LAPC4) and 88% (VCaP) of the serum total androgen receptor transcriptional activity found in castrated cases. The total androgen receptor transcriptional activity secondary to testosterone, dihydrotestosterone and androstenedione was associated with time to castration resistance in patients from the PR.7 study (HR 2.17, 95% CI 1.12-4.23, p=0.02) in multivariate analysis using the castration sensitive model (LAPC4). Androgen receptor transcriptional activity of extragonadal androstenedione was the only steroid statistically associated with time to castration resistance in univariate analysis (HR 1.89, 95% CI 1.04-3.44, p=0.036). CONCLUSIONS: Extragonadal steroids contribute significantly to the androgen receptor axis activation at castration levels of testosterone in recurrent nonmetastatic prostate cancer and these sustain the development of castration resistance after primary local treatment.


Assuntos
Androstenodiona/farmacologia , Castração/métodos , Recidiva Local de Neoplasia/terapia , Neoplasias da Próstata/terapia , Receptores Androgênicos/metabolismo , Testosterona/análogos & derivados , Testosterona/farmacologia , Anabolizantes/farmacologia , Androgênios/farmacologia , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Seguimentos , Humanos , Masculino , Espectrometria de Massas , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Estudos Prospectivos , Próstata/efeitos dos fármacos , Próstata/metabolismo , Próstata/patologia , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Receptores Androgênicos/efeitos dos fármacos , Fatores de Tempo
5.
Cancer Treat Rev ; 81: 101871, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31698174

RESUMO

Acquired resistance to a drug treatment is a common problem across many cancers including prostate cancer (PCa) - one of the major factors for male mortality. The androgen receptor (AR) continues to be the main therapeutic PCa target and despite the success of modern targeted therapies such as enzalutamide, resistance to these drugs eventually develops. The AR has found many ways to adapt to treatments including overexpression and production of functional, constitutively active splice variants. However, of particular importance are point mutations in the ligand binding domain of the protein that convert anti-androgens into potent AR agonists. This mechanism appears to be especially prevalent with the AR in spite of some distant similarities to other hormone nuclear receptors. Despite the AR being one of the most studied and attended targets in cancer, those gain-of-function mutations in the receptor remain a significant challenge for the development of PCa therapies. This drives the need to fully characterize such mutations and to consistently screen PCa patients for their occurrence to prevent adverse reactions to anti-androgen drugs. Novel treatments should also be developed to overcome this resistance mechanism and more attention should be given to the possibility of similar occurrences in other cancers.


Assuntos
Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Neoplasias da Próstata/tratamento farmacológico , Receptores Androgênicos/genética , Antagonistas de Androgênios/farmacologia , Androgênios/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Humanos , Masculino , Terapia de Alvo Molecular , Mutação , Neoplasias da Próstata/genética , Receptores Androgênicos/metabolismo
6.
Yonsei Med J ; 60(12): 1129-1137, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31769243

RESUMO

PURPOSE: Local treatment has become a treatment option for patients with de novo metastatic hormone-sensitive prostate cancer (mHSPC). Subgroup analyses based on a history of cerebrovascular disease (CVD) were performed to evaluate the impact thereof on overall survival (OS) after local treatment. MATERIALS AND METHODS: A retrospective analysis was performed for 879 patients with de novo mHSPC between August 2003 and November 2016. Patients were stratified according to prior CVD history and the type of initial treatment: androgen-deprivation therapy (ADT) alone versus local treatment consisting of radical prostatectomy (RP) or radiation therapy (RT) with ADT, with or without metastasis-directed therapy. The primary outcome was OS assessed by Kaplan-Meier analysis and Cox-regression models. RESULTS: Of 879 patients, 660 (75.1%) men underwent ADT alone, and 219 (24.9%) men underwent RP or RT with ADT, with or without metastasis-directed therapy. The median follow-up was 38 months. Multivariable analysis showed CVD history to be associated with a higher risk of overall mortality (p=0.001). In the overall cohort and in patients without a history of CVD, patients who underwent local treatment exhibited higher OS than men who received ADT alone (all p<0.001). However, the survival benefit conferred by local treatment was not seen in patients with a history of CVD (p=0.324). OS was comparable between patients who received RP and RT (p=0.521). CONCLUSION: Local treatment with or without metastasis-directed therapy may provide OS advantages for mHSPC patients without a history of CVD. Further prospective studies are needed to address these important concerns.


Assuntos
Androgênios/farmacologia , Transtornos Cerebrovasculares/complicações , Estimativa de Kaplan-Meier , Neoplasias da Próstata/patologia , Neoplasias da Próstata/terapia , Idoso , Estudos de Coortes , Humanos , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Modelos de Riscos Proporcionais , Estudos Prospectivos , Estudos Retrospectivos , Fatores de Risco
7.
J Ovarian Res ; 12(1): 82, 2019 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-31472696

RESUMO

Polycystic ovary syndrome (PCOS) is an endocrine disease that is common in women in their reproductive period. Patients with this disease suffer from anovulation and hyperandrogenism. Ovulation induction with exogenous gonadotropin often causes ovarian hyperstimulation syndrome because many small antral follicles pause in their growth. Treatment with insulin sensitizers is reportedly effective for both anovulation associated with PCOS, and suppression of excessive follicular growth; however, the underlying mechanism of action remains unknown. Although pioglitazone is known as an insulin sensitizer, it also has a potent modulator of cell growth and apoptosis irrespective of insulin resistance. To clarify the effect of pioglitazone on follicular growth, we performed in vitro culture of murine preantral follicles. Secondary follicles (100-160 µm in diameter) isolated from 6-week-old ICR mice were individually cultured for 13 days. Culture conditions were as follows: 1) follicle-stimulating hormone (FSH; 33 mIU/mL; control), 2) FSH plus dihydrotestosterone (DHT; 500 ng/mL), 3) FSH plus pioglitazone (5 ng/mL), and 4) FSH plus DHT/pioglitazone. Survival rate and follicle diameter were evaluated, and concentrations of estradiol (E2) and vascular endothelial growth factor (VEGF) in culture media were measured. mRNA expression of various growth-promoting factors and Vegf within follicles were also assessed. Although no significant differences were observed with regard to survival rate, follicle diameters on day 13 were significantly different.Compared with the control group, the DHT group showed enhanced growth, while groups administered pioglitazone showed stagnation of the accelerated growth induced by DHT. Although DHT treatment enhanced the expression of bone morphogenetic protein 2 (Bmp2) mRNA, pioglitazone exposure suppressed induction of Bmp2 mRNA by DHT. Vegf mRNA and protein expression were also significantly reduced when pioglitazone was added to culture media containing DHT.Administration of pioglitazone negatively affected follicular growth and VEGF levels, which may suppress excessive follicular growth and prevent ovarian hyperstimulation syndrome.


Assuntos
Hipoglicemiantes/farmacologia , Folículo Ovariano/efeitos dos fármacos , Pioglitazona/farmacologia , Androgênios/farmacologia , Animais , Proteína Morfogenética Óssea 2/genética , Di-Hidrotestosterona/farmacologia , Estradiol/metabolismo , Feminino , Camundongos Endogâmicos ICR , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
8.
J Steroid Biochem Mol Biol ; 194: 105459, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31470108

RESUMO

The effect of androgen on angiogenesis has been documented. However, its underlying molecular mechanisms have not been well illustrated. Here, we show that treatment with an androgen receptor (AR) agonist, metribolone (R1881; 0.05-5 nM), or dihydrotestosterone (DHT; 0.5-2 nM), concentration- and time-dependently inhibited proliferation in human umbilical venous endothelial cells (HUVEC). This inhibitory effect was confirmed in human microvascular endothelial cells (HMEC-1). Flow cytometric analysis demonstrated that R1881 induced G0/G1 phase cell cycle arrest in HUVEC. Blockade of the AR activity by pre-treatment with an AR antagonist, hydroxyflutamide (HF), or knockdown of AR expression using the shRNA technique abolished the R1881-induced HUVEC proliferation inhibition, suggesting that AR activation can inhibit endothelial cell proliferation. We further investigated the signaling pathway contributing to the proliferation inhibition induced by AR activation. Our data suggest that R1881 reduced the proliferation rate of HUVEC through activating the AR/cSrc/AKT/p38/ERK/NFκB pathway, subsequently up-regulating p53 expression, which in turn increased the levels of p21 and p27 protein, hence decreasing the activities of cyclin-dependent kinase 2 (CDK2) and CDK4, and finally reduced the cell proliferation rate. An extra-nuclear pathway involved in the proliferation inhibition induced by AR activation in vascular endothelial cells was confirmed by showing that membrane-impermeable testosterone-bovine serum albumin (BSA) treatment significantly increased the levels of p53, p27 and p21 protein and reduced cell proliferation. These data highlight the underlying molecular mechanisms by which AR activation induced proliferation inhibition in vascular endothelial cells.


Assuntos
Androgênios/farmacologia , Células Endoteliais/efeitos dos fármacos , Metribolona/farmacologia , Receptores Androgênicos/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , NF-kappa B/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores Androgênicos/genética , Proteína Supressora de Tumor p53/metabolismo
9.
Prostate ; 79(14): 1629-1639, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31376196

RESUMO

BACKGROUND: Recent microarray and sequencing studies of prostate cancer showed multiple molecular alterations during cancer progression. It is critical to evaluate these molecular changes to identify new biomarkers and targets. We performed analysis of glycine-N-acyltransferase like 1 (GLYATL1) expression in various stages of prostate cancer in this study and evaluated the regulation of GLYATL1 by androgen. METHOD: We performed in silico analysis of cancer gene expression profiling and transcriptome sequencing to evaluate GLYATL1 expression in prostate cancer. Furthermore, we performed immunohistochemistry using specific GLYATL1 antibody using high-density prostate cancer tissue microarray containing primary and metastatic prostate cancer. We also tested the regulation of GLYATL1 expression by androgen and ETS transcription factor ETV1. In addition, we performed RNA-sequencing of GLYATL1 modulated prostate cancer cells to evaluate the gene expression and changes in molecular pathways. RESULTS: Our in silico analysis of cancer gene expression profiling and transcriptome sequencing we revealed an overexpression of GLYATL1 in primary prostate cancer. Confirming these findings by immunohistochemistry, we show that GLYATL1 is overexpressed in primary prostate cancer compared with metastatic prostate cancer and benign prostatic tissue. Low-grade cancers had higher GLYATL1 expression compared to high-grade prostate tumors. Our studies showed that GLYATL1 is upregulated upon androgen treatment in LNCaP prostate cancer cells which harbors ETV1 gene rearrangement. Furthermore, ETV1 knockdown in LNCaP cells showed downregulation of GLYATL1 suggesting potential regulation of GLYATL1 by ETS transcription factor ETV1. Transcriptome sequencing using the GLYATL1 knockdown prostate cancer cell lines LNCaP showed regulation of multiple metabolic pathways. CONCLUSIONS: In summary, our study characterizes the expression of GLYATL1 in prostate cancer and explores the regulation of its regulation in prostate cancer showing role for androgen and ETS transcription factor ETV1. Future studies are needed to decipher the biological significance of these findings.


Assuntos
Neoplasias da Próstata/enzimologia , Androgênios/farmacologia , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Imuno-Histoquímica , Masculino , Próstata/enzimologia , Neoplasias da Próstata/patologia , Análise Serial de Tecidos , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia , Sequenciamento Completo do Exoma
10.
J Steroid Biochem Mol Biol ; 195: 105450, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31437548

RESUMO

Estrogen receptor (ER) sequences vary between species and this suggests that there are differences in the ligand-specificity, leading to species-specific effects. This would indicate that it is not possible to generalize effects across species. In this study, we investigated the differences in activation potencies and binding affinities of ER´s alpha (α) and beta (ß) in human, zebrafish and sea bream to elucidate species differences in response to estradiol, estrone, estriol and methyltestosterone. In vitro analysis showed that estradiol had the highest activity for all the ER´s except for human ERß and seabream ERß2. Alignment of the ligand binding domain and ligand binding pocket (LBP) residues of the three species showed that different residues were involved in the LBPs which led to differences in pocket volume, affected binding affinity and orientation of the ligands. By combining in silico and in vitro results, it was possible to identify the ligand specificities of ER´s. The results demonstrated that the human ER´s show lower resolution in ligand-dependent activation, suggesting higher promiscuity, than the zebrafish and seabream ER´s. These results show species-specificity of ER´s and suggest that species-specific differences must be taken into consideration when studying different exposure scenarios.


Assuntos
Androgênios/farmacologia , Estrogênios/farmacologia , Proteínas de Peixes/metabolismo , Receptores Estrogênicos/metabolismo , Animais , Linhagem Celular , Estradiol/farmacologia , Estriol/farmacologia , Estrona/farmacologia , Proteínas de Peixes/genética , Humanos , Ligantes , Metiltestosterona/farmacologia , Simulação de Acoplamento Molecular , Receptores Estrogênicos/genética , Dourada , Especificidade da Espécie , Peixe-Zebra
11.
Physiol Biochem Zool ; 92(5): 459-462, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31365307

RESUMO

Maternal decisions on egg composition have major consequences for offspring. Maternal egg androgens have diverse, often contrasting, effects depending on offspring trait and life stage, suggesting that mothers face trade-offs in egg hormone transfer. However, the effect of egg androgens on embryonic telomere length, which is a major trait potentially affecting performance, has been never investigated. We administered a physiological dose of testosterone (T) to yellow-legged gull (Larus michahellis) eggs and found that, compared to controls, telomere length shortly before hatching was reduced in the liver but unaffected in the brain, heart, and pectoralis muscle. Telomere length varied across somatic tissues, and, independent of egg treatment, it was not correlated between them, suggesting independent telomere dynamics. Thus, we showed for the first time that increased egg T can increase telomere shortening in the embryo and that maternal T allocation strategies may evolve also in response to such effect. Moreover, contrary to observations in adult birds, at the embryonic stage telomere length in one somatic tissue may not reflect telomere length in other body districts.


Assuntos
Androgênios/farmacologia , Charadriiformes/embriologia , Embrião não Mamífero/efeitos dos fármacos , Homeostase do Telômero/efeitos dos fármacos , Testosterona/farmacologia , Androgênios/administração & dosagem , Animais , Óvulo , Telômero , Testosterona/administração & dosagem
12.
Nat Hum Behav ; 3(8): 856-866, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31332298

RESUMO

Moral dilemma judgements frequently involve decisions where moral norms and the greater good are in conflict. The current preregistered study tested the effect of the steroid hormone testosterone on moral dilemma judgements using a double-blind administration of testosterone or placebo. Counter to predictions, testosterone administration led to increased inaction in moral dilemmas where harmful actions prohibited by moral norms increase overall well-being. Using a mathematical model to disentangle sensitivity to consequences, sensitivity to moral norms and general preference for inaction versus action, analyses further revealed that testosterone administration influenced judgements by increasing sensitivity to moral norms. Exploratory analyses suggested the opposite pattern for endogenous testosterone measured at baseline, in that higher levels of endogenous testosterone were associated with lower sensitivity to moral norms. The results indicate that the role of testosterone in moral judgements is more complex than suggested by previous findings. PROTOCOL REGISTRATION: The stage 1 protocol for this Registered Report was accepted in principle on 13 November 2017. The protocol, as accepted by the journal, can be found at https://osf.io/rysbe/ 1.


Assuntos
Androgênios/farmacologia , Tomada de Decisões/efeitos dos fármacos , Teoria Ética , Julgamento/efeitos dos fármacos , Princípios Morais , Testosterona/farmacologia , Adolescente , Método Duplo-Cego , Feminino , Humanos , Masculino , Normas Sociais , Adulto Jovem
13.
J Steroid Biochem Mol Biol ; 192: 105406, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31185279

RESUMO

Prostate cancer (PCa) is one of the most common malignancies and the second most common cause of cancer-related deaths in men world-wide and is known to be affected by the action of dihydrotestosterone (DHT) via androgen receptor (AR). Resveratrol (Res) as a phytochemical in grapes and red wine has diverse biological effects such as anti-inflammation, anti-oxidation and anti-cancer. CXCR4 as a chemokine receptor has been found to be upregulated in cancer metastasis and has been used as a prognostic marker in various types of cancer, including leukemia, breast cancer, and prostate cancer. In this study, we focused on the role of DHT in the induction of prostate cancer progression by affecting the AR and CXCR4 pathway. Also, we investigated the inhibition effect of resveratrol on DHT-induced prostate cancer metastasis. In cell viability assay, DHT increased the cell viability of LNCaP prostate cancer cells, on the other hand, Res and its combination with bicalutamide (BCT) as an AR-antagonist or AMD3100 as a CXCR4 inhibitor significantly reduced the cell viability promoted by DHT. Trans-well migration assay and wound healing assay represented the similar results with cell viability assay. According to the results of TUNEL assay, the apoptotic activity was induced by treatment of Res. As results of western blot analysis, the expression of AR, CXCR4, p-PI3K, and p-AKT and the downstream genes related with cell cycle progression and epithelial-mesenchymal transition (EMT) were decreased and the expression of the apoptosis-related genes was increased by treatment of Res and its combination with BCT or AMD3100. This study would suggest that Res and its combination with AR and CXCR4 antagonists can be used in order to suppress the metastatic behaviors of prostate cancer.


Assuntos
Antagonistas de Receptores de Andrógenos/farmacologia , Di-Hidrotestosterona/farmacologia , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Neoplasias da Próstata/tratamento farmacológico , Receptores Androgênicos/química , Receptores CXCR4/antagonistas & inibidores , Resveratrol/farmacologia , Androgênios/farmacologia , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Progressão da Doença , Humanos , Masculino , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Transdução de Sinais , Células Tumorais Cultivadas
14.
J Physiol Pharmacol ; 70(1)2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31172974

RESUMO

Within the mammalian reproductive system sirtuin 1 and 6 (SIRT1, SIRT6) are considered to contribute to steroid hormone signaling and control of reproductive physiology. Therefore, the specific question is whether and how a commonly used dicarboximide fungicide with antiandrogenic activity, vinclozolin (Vnz) alters SIRT1 and SIRT6 expression and whether both investigated sirtuins positively affect survival of the follicles after vinclozolin exposure. Immunocytochemistry and immunohistochemistry were performed to localize SIRT1 and SIRT6 expression in cultured granulosa cells (GCs; 48 hours) and whole ovarian follicles (24 hours) after treatment with two androgens, testosterone (T; 10-7 M) and dihydrotestosterone (DHT; 10-7 M), and an antiandrogen, Vnz (1.4 x 10-5 M), separately and in combinations. Granulosal and follicular mRNA and protein expression of both sirtuins was also investigated by real-time PCR and Western blot. In addition, their concentration and activity was studied by immunoenzymatic and fluorescence assays. Our observations: (1) demonstrate the presence of both investigated sirtuins in ovarian cells, (2) show their potential involvement in the control of follicular atresia because of increased SIRT1/SIRT6 expression and SIRT1 activity after exposure to Vnz, (3) represent the first data on the interrelationships between sirtuins and androgens in porcine ovarian cells. Based on these findings and our previous results we can conclude, that SIRT1 and SIRT6 do not exert the protective effects in ovarian follicles after vinclozolin exposure. These novel data on the role of SIRT1/SIRT6 in porcine ovarian follicles shows that in the presence of the investigated fungicide, sirtuins are upregulated, which can induce apoptosis of follicular cells. Furthermore the androgen receptor sensitivity to ligands, especially environmental ones (for example: vinclozolin) might be directly linked with the mechanism of action of both investigated sirtuins in the porcine ovary, which requires further investigation.


Assuntos
Antagonistas de Androgênios/farmacologia , Ovário/efeitos dos fármacos , Oxazóis/farmacologia , Sirtuínas/metabolismo , Androgênios/farmacologia , Animais , Di-Hidrotestosterona/farmacologia , Feminino , Ovário/metabolismo , Sirtuínas/genética , Suínos , Testosterona/farmacologia
15.
Toxicol In Vitro ; 60: 203-211, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31154061

RESUMO

The development and normal function of prostate tissue depends on signalling interactions between stromal and epithelial compartments. Development of a prostate microtissue composed of these two components can help identify substance exposures that could cause adverse effects in humans as part of a non-animal risk assessment. In this study, prostate microtissues composed of human derived stromal (WPMY-1) and epithelial (RWPE-1) cell lines grown in scaffold-free hydrogels were developed and characterized using immunohistochemistry, light microscopy, and qRT-PCR. Within 5 days after seeding, the microtissues self-organized into spheroids consisting of a core of stromal WPMY-1 cells surrounded by epithelial RWPE-1 cells. The RWPE-1 layer is reflective of intermediate prostatic epithelium, expressing both characteristics of the luminal (high expression of PSA) and basal (high expression of cytokeratins 5/6 and 14) epithelial cells. The response of the microtissues to an androgen (dihydrotestosterone, DHT) and an anti-androgen (flutamide) was also investigated. Treatment with DHT, flutamide or a mixture of DHT and flutamide indicated that the morphology and self-organization of the microtissues is androgen dependent. qRT-PCR data showed that a saturating concentration of DHT increased the expression of genes coding for the estrogen receptors (ESR1 and ESR2) and decreased the expression of CYP1B1 without affecting the expression of the androgen receptor. With further development and optimization RWPE-1/WPMY-1 microtissues can play an important role in non-animal risk assessments.


Assuntos
Alternativas aos Testes com Animais , Próstata , Antagonistas de Androgênios/farmacologia , Androgênios/farmacologia , Linhagem Celular , Técnicas de Cocultura , Citocromo P-450 CYP1B1/genética , Di-Hidrotestosterona/farmacologia , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Flutamida/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hidrogéis , Masculino , Receptores Androgênicos/genética
16.
Gynecol Endocrinol ; 35(8): 669-672, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31056990

RESUMO

Hyperandrogenism is one of the most common causes for anovulation in women and increases the risk for metabolic disorder in PCOS patients. Autophagy plays an important role in dysfunction of endocrine and anovulation. However, the relationship between hyperandrogenism and autophagy in human granulosa cells of PCOS patients remains unclear. By collecting granulosa cells from PCOS patients and non-PCOS patients, we found that the abundance of autophagy-related genes ATG5, ATG7, BECN1 mRNA and the ratio of autophagy marker protein light chain 3B II/I (LC3 II/I) were significantly increased whereas the abundance of the autophagy substrate SQSTM1/p62 was decreased in ovarian granulosa cells from PCOS patients. Furthermore, we demonstrated that BECN1 mRNA abundance in human granulosa cells positively correlated with the basal level of serum total testosterone and androgen up-regulated the abundance of BECN1 mRNA and the ratio of LC3II/LC3I in a dose-dependent manner in cultured granulosa cells. These observations indicated that androgen-induced activation of autophagy may play an important role in the development of PCOS and to explore the autophagy mechanisms involved in PCOS yield new insight into the pathophysiology and therapy of the disorder.


Assuntos
Androgênios/fisiologia , Autofagia/fisiologia , Células da Granulosa/fisiologia , Síndrome do Ovário Policístico/patologia , Adulto , Androgênios/metabolismo , Androgênios/farmacologia , Autofagia/efeitos dos fármacos , Autofagia/genética , Estudos de Casos e Controles , Células Cultivadas , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Células da Granulosa/patologia , Humanos , Hiperandrogenismo/complicações , Hiperandrogenismo/metabolismo , Hiperandrogenismo/patologia , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/metabolismo , Cultura Primária de Células , Adulto Jovem
17.
Endocrinology ; 160(7): 1757-1770, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31074799

RESUMO

Although prominent sex differences exist in the hypothalamic-pituitary-adrenal axis's response to stressors, few studies of its regulation in the hypothalamic paraventricular nucleus (PVN) have compared both male and female subjects. In this study, we sought to explore sex differences in the acute regulation of PVN neuropeptide expression following glucocorticoid (GC) removal and the underlying role of gonadal hormones. We first examined the effects of short-term adrenalectomy (ADX) on PVN Crh and arginine vasopressin (Avp) expression in mice using in situ hybridization. ADX increased PVN AVP mRNA levels in both sexes. In contrast, PVN CRH mRNA was increased by 2 days after ADX in males only. Both sexes showed increases in CRH mRNA after 4 days. To determine if gonadal hormones contributed to this sex bias, we examined adrenalectomized (ADX'd) and gonadectomized (GDX'd) mice with or without gonadal hormone replacement. Unlike the pattern in intact animals, 2 days following ADX/gonadectomy, CRH mRNA levels did not increase in either sex. When males were given DHT propionate, CRH mRNA levels increased in ADX'd/GDX'd males similar to those observed following ADX alone. To determine a potential mechanism, we examined the coexpression of androgen receptor (AR) immunoreactivity and CRH neurons. Abundant colocalization was found in the anteroventral bed nucleus of the stria terminalis but not the PVN. Thus, our findings reveal a sex difference in PVN Crh expression following the removal of GC-negative feedback that may depend on indirect AR actions in males.


Assuntos
Adrenalectomia , Androgênios/metabolismo , Hormônio Liberador da Corticotropina/metabolismo , Hipotálamo/metabolismo , Receptores Androgênicos/metabolismo , Androgênios/farmacologia , Animais , Arginina Vasopressina/genética , Arginina Vasopressina/metabolismo , Castração , Hormônio Liberador da Corticotropina/genética , Di-Hidrotestosterona/farmacologia , Feminino , Hipotálamo/efeitos dos fármacos , Masculino , Camundongos , Neurônios/metabolismo , Fatores Sexuais
18.
Int J Hyg Environ Health ; 222(4): 670-677, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31129136

RESUMO

Endocrine active substances (EAS), which are commonly used in pharmaceuticals and personal care products, are released into surface water mainly through WWTP effluents and have been shown to cause adverse effects in aquatic organisms. In wastewater, a variety of EAS with different hormonal activities is present, which can lead to additive effects or mask an endocrine activity. To investigate hormonal combination effects, with a focus on estrogen and androgen-modulators, influent samples from municipal and hospital wastewater treatmenr plants were spiked with 17α-ethinylestradiol, toremifene, 17α-methyltestosterone and bicalutamide and analyzed using in vitro reporter gene CALUX assays. All wastewaters caused endocrine activities in human cells, which were modified by adding one or several endocrine active substances. As expected, estrogenic activity was reduced in presence of the anti-estrogenic toremifene and androgenic activity decreased with the anti-androgen bicalutamide. In general, substance addition caused a similar trend in altered endocrine activities; however, their intensities differed between the wastewaters. Our results indicate that masking effects, leading to a suppressed biological signal, are of significant importance in the assessment of complex water samples, and combination effects rather than single substances determine the final biological effect. This emphasizes the need of effect-based tools in the assessment of water samples.


Assuntos
Androgênios/farmacologia , Disruptores Endócrinos/farmacologia , Estrogênios/farmacologia , Águas Residuárias , Poluentes Químicos da Água/farmacologia , Anilidas/farmacologia , Bioensaio , Linhagem Celular Tumoral , Interações Medicamentosas , Etinilestradiol/farmacologia , Genes Reporter , Humanos , Luciferases/genética , Metiltestosterona/farmacologia , Nitrilos/farmacologia , Receptores Androgênicos/metabolismo , Receptores Estrogênicos/metabolismo , Toremifeno/farmacologia , Compostos de Tosil/farmacologia
19.
Int J Mol Sci ; 20(9)2019 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-31052484

RESUMO

Regioselective synthesis of novel ring A-fused arylpyrazoles of dihydrotestosterone (DHT) was carried out in two steps under facile reaction conditions. Aldol condensation of DHT with acetaldehyde afforded a 2-ethylidene derivative regio- and stereo-selectively, which was reacted with different arylhydrazines in the presence of iodine via microwave-assisted oxidative cyclization reactions. The 17-keto analogs of steroidal pyrazoles were also synthesized by simple oxidation in order to enlarge the compound library available for pharmacological studies and to obtain structure-activity relationship. The antiproliferative activities of the structurally related heteroaromatic compounds were tested in vitro on human cervical and breast adenocarcinoma cell lines (HeLa, MCF-7 and MDA-MB-231) and on two androgen-independent malignant prostate carcinoma cell lines (PC-3 and DU 145). Based on primary cytotoxicity screens and IC50 assessment, a structure-function relationship was identified, as derivatives carrying a hydroxyl group on C-17 exhibit stronger activity compared to the 17-one counterparts. Cancer cell selectivity of the derivatives was also determined using non-cancerous MRC-5 cells. Furthermore, the proapoptotic effects of some selected derivatives were verified on androgen therapy refractive p53-deficient PC-3 cells. The present study concludes that novel DHT-derived arylpyrazoles exert cancer cell specific antiproliferative activity and activate apoptosis in PC-3 cells.


Assuntos
Antineoplásicos/química , Antineoplásicos/farmacologia , Di-Hidrotestosterona/química , Di-Hidrotestosterona/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Pirazóis/química , Pirazóis/farmacologia , Androgênios/farmacologia , Antineoplásicos/síntese química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Di-Hidrotestosterona/síntese química , Humanos , Masculino , Modelos Moleculares , Pirazóis/síntese química
20.
Horm Behav ; 113: 13-20, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31054274

RESUMO

Anabolic-androgenic steroids (AAS) are drugs of abuse that impair behavior and cognition. In a rodent model of AAS abuse, testosterone-treated male rats expend more physical effort, by repeatedly pressing a lever for a large reward in an operant discounting task. However, since modern society prioritizes cognitive over physical effort, it is important to determine if AAS limit cognitive effort. Here we tested the effects of AAS on a novel cognitive-effort discounting task. Each operant chamber had 3 nose-pokes, opposite 2 levers and a pellet dispenser. Rats pressed a lever to illuminate 1 nose-poke; they responded in the illuminated nose-poke to receive sugar pellets. For the 'easy' lever, the light remained on for 1 s, and a correct response earned 1 pellet. For the 'hard' lever, the light duration decreased from 1 s to 0.1 s across 5 blocks of trials, and a correct response earned 4 pellets. As the duration of the nose-poke light decreased, all rats decreased their choice of the hard lever in a modest discounting curve. Task accuracy also decreased significantly across the 5 blocks of trials. However, there was no effect of testosterone on choice of the hard lever or task accuracy. Antagonism of dopamine D1 or D2 receptors had no effect on lever choice or task accuracy. However, serotonin depletion significantly decreased preference for the hard lever, and impaired task accuracy. Thus, physical effort discounting depends on dopamine activity, while cognitive effort discounting task is sensitive to serotonin. AAS impair physical effort discounting, but not cognitive effort discounting.


Assuntos
Anabolizantes/farmacologia , Androgênios/farmacologia , Cognição/efeitos dos fármacos , Condicionamento Operante/efeitos dos fármacos , Esteroides/farmacologia , Animais , Dopamina/farmacologia , Dopamina/fisiologia , Antagonistas de Dopamina/farmacologia , Masculino , Ratos , Ratos Long-Evans , Receptores de Dopamina D2/efeitos dos fármacos , Receptores de Dopamina D2/metabolismo , Recompensa , Testosterona/farmacologia
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