Influence of CYP19A1 gene expression levels in women with breast cancer: a systematic review of the literature

Breast cancer is the most frequently diagnosed malignant neoplasm in women and is considered a multifactorial disease of unknown etiology. One of the major risk factors is genetic alteration. Changes in CYP19A1 gene expression levels have been associated with increased risk and increased aggressiveness of breast cancer. Increased CYP19A1 gene expression and/or aromatase activity are among the major regulatory events for intratumoral production of estrogens in breast malignant tissues. This systematic review aimed to investigate the influence of CYP19A1 gene expression levels in women with breast cancer. The research was carried out using the PubMed, Scopus, and Web of Science databases. Searches were conducted between February 2 and May 15, 2019. Inclusion criteria were studies published between 2009 and 2019, English language publications, and human studies addressing the gene expression of CYP19A1 in breast cancer. A total of 6.068 studies were identified through PubMed (n=773), Scopus (n=2,927), and the Web of Science (n=2,368). After selecting and applying the inclusion and exclusion criteria, six articles were included in this systematic review. This systematic review provides evidence that increased or decreased levels of CYP19A1 gene expression may be related to pathological clinical factors of disease, MFS, OS, DFS, WATi, markers of metabolic function, concentrations of E1, FSH, and in the use of multiple exons 1 of the CYP19A1 gene in breast cancer.

Pretentious genomic selection signatures in CYP19A1 gene associated with silent estrous behavior in water buffalo in Pakistan

Background: Poor reproductive efficiency of river buffalos hampers the production capabilities of animals. Buffalos are mainly considered poor breeders owing to the constrained expression of estrus behavior. Failure to display heat signs is an indication of improper functionality of signaling peptides to trigger on a series of behavioral changes, which can be detectable by breeders for timely insemination of females. This might cause an animal to be a repeat breeder. Genomic variations underlying synthesis of signaling peptides can be a useful marker to select superior animals with better reproductive efficiency. In this context, the current study was designed to analyze the CYP19A1 gene in Nili-Ravi buffalo. Results: A total of 97 animals were selected and were divided into two groups on the basis of their heat score. PCR amplification and sequencing of the amplicons were performed using the specific sets of primer, and then, sequences were analyzed for novel variants. A total of 11 polymorphic sites were identified illustrating phenotypic variation in the heat score. Most of the loci were found homologous. Single Nucleotide Polymorphisms (SNPs) were analyzed for association with silent estrus. A three-dimensional protein model was also generated to locate the position of exonic SNPs. Conclusion: This study illustrated that polymorphic sites in the CYP19A1 gene provided potential markers for selection of buffalos with better estrus behavior.

Combined Effect of the PGR + 331C > T, CYP17A1 -34A > G and CYP19A1 1531G > A Polymorphisms on the Risk of Developing Endometriosis / Efeito combinado dos polimorfismos PGR + 331C > T, CYP17A1 -34A > G e CYP19A1 1531G > A no risco de desenvolvimento da endometriose

Abstract Purpose To evaluate the magnitude of the association of the polymorphisms of the genes PGR, CYP17A1 and CYP19A1 in the development of endometriosis. Methods This is a retrospective case-control study involving 161 women with endometriosis (cases) and 179 controls. The polymorphisms were genotyped by real-time polymerase chain reaction using the TaqMan system. The association of the polymorphisms with endometriosis was evaluated using the multivariate logistic regression. Results The endometriosis patients were significantly younger than the controls (36.0±7.3 versus 38.0±8.5 respectively, p = 0.023), and they had a lower body mass index (26.3±4.8 versus 27.9±5.7 respectively, p = 0.006), higher average duration of the menstrual flow (7.4±4.9 versus 6.1±4.4 days respectively, p = 0.03), and lower average time intervals between menstrual periods (25.2±9.6 versus 27.5±11.1 days respectively, p = 0.05). A higher prevalence of symptoms of dysmenorrhea, dyspareunia, chronic pelvic pain, infertility and intestinal or urinary changes was observed in the case group when compared with the control group. The interval between the onset of symptoms and the definitive diagnosis of endometriosis was 5.2±6.9 years. When comparing both groups, significant differences were not observed in the allelic and genotypic frequencies of the polymorphisms PGR + 331C > T, CYP17A1 -34A > G and CYP19A1 1531G > A, even when considering the symptoms, classification and stage of the endometriosis. The combined genotype PGR + 331TT/CYP17A1 -34AA/CYP19A11531AA is positively associated with endometriosis (odds ratio [OR] = 1.72; 95% confidence interval [95%CI] = 1.09-2.72). Conclusions The combined analysis of the polymorphisms PGR-CYP17A1-CYP19A1 suggests a gene-gene interaction in the susceptibility to endometriosis. These results may contribute to the identification of biomarkers for the diagnosis and/or prognosis of the disease and of possible molecular targets for individualized treatments.

Resumo Objetivo Avaliar a magnitude de associação de polimorfismos nos genes PGR, CYP17A1 e CYP19A1 no desenvolvimento da endometriose. Métodos Este é um estudo retrospectivo do tipo caso-controle, envolvendo 161 mulheres com endometriose (casos) e 179 controles. Os polimorfismos foram genotipados pela reação em cadeia da polimerase em tempo real utilizando o sistema TaqMan. A associação dos polimorfismos estudados com a endometriose foi avaliada pela regressão logística multivariada. Resultados As pacientes com endometriose eram significativamente mais jovens do que os controles (36,0±7,3 versus 38,0±8,5, respectivamente, p = 0,023), apresentaram um índice de massa corporal menor (26,3±4,8 versus 27,9±5,7, respectivamente, p = 0,006), maior tempo médio de duração do fluxo menstrual (7,4±4,9 versus 6,1±4,4 dias, respectivamente, p = 0,03) e menor tempo médio do intervalo entre as menstruações (25,2±9,6 versus 27,5±11,1 dias, respectivamente, p = 0,05). Uma maior prevalência dos sintomas de dismenorreia, dispareunia, dor pélvica crônica, infertilidade, alterações intestinais e urinárias foi observada no grupo casos comparado ao grupo controle. O tempo médio entre o início dos sintomas e o diagnóstico definitivo de endometriose foi de 5,2±6,9 anos. Comparando os dois grupos, não foram observadas diferenças significativas nas frequências alélicas e genotípicas dos polimorfismos PGR + 331C > T, CYP17A1 -34A > G e CYP19A1 1531G > A, e nem considerando os sintomas, a classificação e o estadiamento da endometriose. O genótipo combinado PGR + 331TT/CYP17A1 -34AA/CYP19A11531AA está associado positivamente com a endometriose (razão de possibilidades [RP] = 1,72; intervalo de confiança de 95% [IC95%] = 1,09-2,72). Conclusões A análise combinada dos polimorfismos PGR-CYP17A1-CYP19A1 sugere uma interação gene-gene na susceptibilidade à endometriose. Estes resultados podem contribuir para a identificação de biomarcadores para o diagnóstico e/ou prognóstico da doença, assim como de possíveis alvos moleculares para um tratamento individualizado.

Androgênios e mama / Androgens and breast

O câncer de mama (BCA) é uma das neoplasias mais frequentes em mulheres de vários países, a exposição excessiva aos estrogênios é um dos principais fatores de risco. Os ovários são as principais fontes de produção estrogênica endógena; porém, na menopausa essa produção cessa e a síntese extragonadal, sobretudo nas células mesenquimais do tecido adiposo, passa a ser a principal fonte de produção estrogênica, pois essas células apresentam aromatase, enzima que converte androgênios em estrogênios. Apoiada por fortes evidências clínicas, a reposição androgênica tem sido recomendada para o alívio de sintomas decorrentes da síndrome da insuficiência androgênica feminina, tais como fadiga, alterações do humor e quadros de depressão; além disso, estudos experimentais têm sugerido a possibilidade de uma plausível proteção da reposição androgênica contra o BCA. Nesses estudos, em que atuou por meio de seus receptores, a testosterona apresentou efeitos antiproliferativos, pró­apoptóticos e inibiu a atividade dos receptores estrogênicos e do crescimento de tumores mamários; evidências clínicas também apoiam o papel protetor dos androgênios na mama. Entretanto, outros estudos indicam que esse papel protetor depende do nível de atividade da aromatase; assim, a testosterona pode exercer um efeito inibidor direto no crescimento tumoral ao ligar­se ao seu receptor, porém ter um efeito estimulador indireto através de sua conversão para estrogênios pela aromatase. A obesidade e a insulina, além de múltiplos outros fatores, alguns dos quais são fatores de risco independentes para BCA, podem resultar na superexpressão da aromatase e ter como resultado aumento na produção localizada de estrogênios, os quais são fatores estimulantes do BCA. Estudos sobre a administração de testosterona em mulheres são escassos e controversos e não existem estudos que forneçam dados em termos de segurança desse uso em longo prazo. Assim, nesta revisão pretendemos mostrar como os androgênios atuam na mama. Frente às evidências atuais, o uso de androgênios em mulheres com fatores de risco para câncer de mama não é recomendado.(AU)

Breast cancer (BCAA) is one of the most frequent malignancies in women in several countries, which excessive exposure to oestrogens is one of the main risk factors. The ovaries are the main source of endogenous estrogen production; however, at menopause this production sessate and extra­gonadal synthesis, especially in ectomesenchymal cells from adipose tissue, turns the main source of estrogen production, since these cells express aromatase, an enzyme that converts androgens to estrogens. Supported by strong clinical evidence androgen replacement has been recommended for the relief of symptoms caused by female syndrome of androgen insufficiency, such as fatigue, mood swings and depression; Furthermore, experimental studies have suggested the possibility of protection of androgen replacement against BCA. In these studies, acting through their receptors, testosterone showed antiproliferative, proapoptotic and inhibited the activity of estrogen receptors and growth of mammary tumors; Clinical evidence also support the protective role of androgens in the breast. However, studies indicate that this protective role depends on the level of aromatase activity; for instance, testosterone can exert a direct inhibitory effect on tumor growth by binding to its receptor, but have an indirect effect by stimulating its conversion to oestrogens by aromatase. Obesity and insulin, as well as multiple other factors, some of which are independent risk factors for BCA, may result in overexpression of aromatase, resulting in increased localized production of estrogens, which are inducible factors of BCA. Studies on the administration of testosterone in women are scarce and controversial, and there are no studies that provide data in terms of long­term use of safety. Thus, in this review we intend to show how androgens act in the breast. Given the current evidence, the use of androgens in women with risk factors for breast cancer is not recommended.(AU)

Polymorphism C242T in the Cyp19 gene in meat sheep / Polimorfismo no gene cyp19 em ovinos de corte

Abstract The objective of this study was to evaluate the frequency of C242T polymorphism on the aromatase gene and the allelic and genotypic frequency of these variants in sheep belonging to four breed groups. Blood samples were collected from 187 animals of four breed groups: Dorper, Santa Inês, Texel and White Dorper, originated from herds in the region of Maringá/PR, Brazil. The genomic DNA was extracted using alkaline extraction, with subsequent amplification of the fragments via PCR with specific primer. The samples resulting from amplification were subjected to digestion process using the Dpn II restriction enzyme and to polyacrylamide gel electrophoresis 10.0% and stained with silver nitrate. Three distinct genotypes were observed: homozygous (CC), heterozygous (CT) and homozygous for no cut (TT). The resulting data were analyzed using the POPGENE software with 5% significance. Genotypic frequencies among the breed groups were: Texel (CC - 0.426; CT - 0.511; TT - 0.064), Dorper (CC - 0.073; CT - 0.732; TT - 0.439), White Dorper (CC - 0.021; CT - 0.255; TT - 0.723) and Santa Inês (CC - 0.115; CT - 0.462; TT - 0.423).

Resumo O objetivo deste trabalho foi avaliar as frequências alélicas e genotípicas do polimorfismo do C242T no gene da aromatase em ovinos pertencentes a quatro grupos raciais. Foram coletadas amostras de sangue de 187 animais de quatro grupos raciais: Dorper, Santa Inês, Texel e White Dorper, provenientes de rebanhos da região de Maringá, PR - BR. O DNA genômico foi extraído utilizando o método de extração alcalina, com posterior amplificação dos fragmentos via PCR com primer específico. As amostras resultantes da amplificação foram submetidas ao processo de digestão com auxilio da enzima restrição Dpn II e submetido à eletroforese em gel de poliacrilamida de 10,0% e corado nitrato de prata. Foram observados três genótipos distintos: Homozigoto (CC), heterozigoto (CT) e homozigoto para não corte (TT). Os dados resultantes foram analisados utilizando o software POPGENE com significância de 5%. As frequências genotípicas entre os grupos raciais foram: Texel (CC - 0,426; CT - 0,511; TT - 0,064), Dorper (CC - 0,073; CT - 0,732; TT - 0,439), White Dorper (CC - 0,021; CT - 0,255; TT - 0,723) e Santa Inês (CC - 0,115; CT - 0,462; TT - 0,423).

Aspectos genéticos relacionados ao transexualismo / Genetic aspects related to transsexualism

Transexual é quem não se identifica com o seu sexo de nascimento e que procura adequar ou passou por adequação para o gênero desejado, o que em vários, mas não em todos os casos, envolve transição somática por tratamento hormonal e cirurgia genital (cirurgia de redesignação sexual).O envolvimento de fatores genéticos no transexualismo é oriundo, sobretudo, de estudos familiares, casos de gêmeos em concordância para a transexualidade e a partir de pesquisas da genética molecular de certos polimorfismos do sistema de genes androgênicos e estrogênicos.O receptor androgênico (RA) é responsável pela diferenciação no córtex cortical e está associado com sua masculinização na adolescência.Foram identificados dois subtipos de receptores estrogênicos: o ER? e o ER?. O subtipo beta é claramente mais expressivo em várias regi-ões cerebrais. O receptor ER? está primariamente envolvido na masculinização, enquanto o ER? tem papel predominante na ?defeminiza-ção? dos comportamentos sexuais. Uma pesquisa com 29 mulheres transexuais identificou associação significativa com polimorfismos dos nucleotídeos CA no ER? (P = 0,03). Em 112 mulheres transexuais foi encontrada uma relação entre polimorfismos do gene longo do AR e transexualismo (P = 0,04). Duzentos e setenta e três homens transexuais, diferiam, significativamente, dos controles na repetição do tamanho médio do polimorfismo dos ER? (P = 0,002).

Differential expression of upstream stimulatory factor (USF) 2 variants in eutopic endometria from women with endometriosis: estradiol regulation

BACKGROUND: Endometriosis, pro-inflammatory and invasive benign disease estrogen dependent, abnormally express in endometria the enzyme P450Arom, positively regulated by steroid factor-1 (SF-1). Our objective was to study the nuclear protein contents of upstream stimulating factor 2 (USF2a and USF2b), a positive regulator of SF-1, throughout the menstrual cycle in eutopic endometria from women with and without (control) endometriosis and the involvement of nuclear estrogen receptors (ER) and G-coupled protein estrogen receptor (GPER)-1. RESULTS: Upstream stimulating factor 2 protein contents were higher in mid (USF2b) and late (USF2a and USF2b) secretory phase in eutopic endometria from endometriosis than control (p < 0.05). In isolated control epithelial cells incubated with E2 and PGE2, to resemble the endometriosis condition, the data showed: (a) significant increase of USF2a and USF2b nuclear protein contents when treated with E2, PPT (specific agonist for ERa) or G1 (specific agonist for GPER1); (b) no increase in USF2 binding to SF-1 E-Box/DNA consensus sequence in E2-treated cells; (c) USF2 variants protein contents were not modified by PGE2; (d) SF-1 nuclear protein content was significantly higher than basal when treated with PGE2, E2 or G1, stimulation unaffected by ICI (nuclear ER antagonist); and (e) increased (p < 0.05) cytosolic protein contents of P450Arom when treated with PGE2, E2, PPT or G1 compared to basal, effect that was additive with E2 + PGE2 together. Nevertheless, in endometriosis cells, the high USF2, SF-1 and P450Arom protein contents in basal condition were unmodified. CONCLUSION: These data strongly suggest that USF2 variants and P450Arom are regulated by E2 through ERa and GPER1, whereas SF-1 through GPER1, visualized by the response of the cells obtained from control endometria, being unaffected the endogenously stimulated cells from endometriosis origin. The lack of E2 stimulation on USF2/SF-1 E-Box/DNA-sequence binding and the absence of PGE2 effect on USF2 variants opposite to the strong induction that they exert on SF1 and P450 proteins suggest different mechanisms and indirect regulations. The sustained USF2 variants protein expression during the secretory phase in eutopic endometria from women with endometriosis may participate in the pathophysiology of this disease strongly associated with infertility and its characteristic endometrial invasion to ectopic sites in the pelvic cavity.

Culture of bovine ovarian follicle wall sections maintained the highly estrogenic profile under basal and chemically defined conditions

Follicle cultures reproduce in vitro the functional features observed in vivo. In a search for an ideal model, we cultured bovine antral follicle wall sections (FWS) in a serum-free defined medium (DM) known to induce 17β-estradiol (E2) production, and in a nondefined medium (NDM) containing serum. Follicles were sectioned and cultured in NDM or DM for 24 or 48 h. Morphological features were determined by light microscopy. Gene expression of steroidogenic enzymes and follicle-stimulating hormone (FSH) receptor were determined by RT-PCR; progesterone (P4) and E2 concentrations in the media were measured by radioimmunoassay. DM, but not NDM, maintained an FWS morphology in vitro that was similar to fresh tissue. DM also induced an increase in the expression of all steroidogenic enzymes, except FSH receptor, but NDM did not. In both DM and NDM, there was a gradual increase in P4 throughout the culture period; however, P4 concentration was significantly higher in NDM. In both media, E2 concentration was increased at 24 h, followed by a decrease at 48 h. The E2:P4 ratio was higher in DM than in NDM. These results suggest that DM maintains morphological structure, upregulates the expression of steroidogenic enzyme genes, and maintains steroid production with a high E2:P4 ratio in FWS cultures.

Mulheres inférteis com endometriose pélvica mínima e leve submetidas à estimulacão ovariana apresentam menor expressão do gene CYP19A1 em células do cumulus / Infertile women with minimal and mild endometriosis submitted tocontrolled ovarian stimulation present lower expression of the gene CYP19A1 in cumulus cells

Objetivo: Menores taxas de gestação em portadoras de endometriose submetidas a técnicas de reprodução assistida podem estar relacionadas à piora da qualidade oocitária. A análise da expressão gênica em células do cumulus (CC) pode fornecer biomarcadores passíveis de predizer a qualidade gamética. O objetivo deste estudo foi comparar os níveis da expressão do gene CYP19A1 em CC de mulheres inférteis com endometriose mínima/leve (I/II) e controles inférteis. Método: Foram selecionadas pacientes com infertilidade por endometriose pélvica inicial e por fator masculino e/ou tubário (grupo controle), submetidas à estimulação ovariana controlada para injeção intracitoplasmática de espermatozoide (ICSI). Imediatamente após a captação oocitária, CC foram isoladas e armazenadas. Foi realizada a quantificação da expressão do gene CYP19A1 nas CC por meio de PCR-Real Time. Resultados: Foram isoladas CC de 23 mulheres inférteis com endometriose I/II e de 41 controles. Observou-se expressão significativamente menor do gene CYP19A1 em CC de mulheres inférteis com endometriose I/II (0,56 ± 0,17) quando comparadas às controles (0,15 ± 0,04) (p = 0,043). Conclusões: A menor expressão do gene CYP19A1 em CC de mulheres inférteis com endometriose pélvica em estágios iniciais pode mediar a piora da qualidade oocitária, abrindo novas perspectivas no entendimento da etiopatogênese da infertilidade relacionada à doença.

Objective: Lower pregnancy rate in women with endometriosis submitted to assisted reproductive techniques might be related to poor oocyte quality. The analysis of the expression of the genes in cumulus cells (CC) might provide biomarkers that can predict gamete quality. The main objective of the present study was to compare the levels of the expression of the gene CYP19A1 in CC of infertile women with minimal and mild (I/II) endometriosis and infertile controls. Method: There were selected patients with infertility caused by initial pelvic endometriosis and by male/tubal factor (control group), submitted to controlled ovarian stimulation to ICSI. Immediately after the oocyte retrieval, CC were isolated and stored. Quantification of the expression of the gene CYP19A1 in CC was performed using PCR-real time.Results: CC were isolated from 23 infertile women with endometriosis I/II and 41 from control. Significant lower expression of the gene CYP19A1 in CC was observed in infertile women with endometriosis I/II (0.56 ± 0.17) when compared to control (0.15 ± 0.04) (p = 0,043). Conclusions: The lower expression of the gene CYP19A1 in CC of infertile women with pelvic endometriosis in initial stages might mediate the poor oocyte quality, opening new perspectives on the understanding of the etiopathogenesis of infertility related to the disease.

Triorganotin as a compound with potential reproductive toxicity in mammals

Organotin compounds are typical environmental contaminants and suspected endocrine-disrupting substances, which cause irreversible sexual abnormality in female mollusks, called "imposex". However, little is known about the capability of triorganotin compounds, such as tributyltin and triphenyltin, to cause disorders in the sexual development and reproductive functions of mammals, including humans and rodents. Moreover, these compounds can act as potential competitive inhibitors of aromatase enzyme and other steroidogenic enzymes, affecting the reproductive capacity of male and female mammals. In this review, we discuss the cellular, biochemical, and molecular mechanisms by which triorganotin compounds induce adverse effects in the mammalian reproductive function.

Análise da expressão de receptores hormonais (androgênio, estrogênio alfa e beta) e da aromatase em carcinomas epidermóides de boca (CE) / Analysis of the expression of hormonal receptors (androgen, estrogen alpha and beta) and aromatase enzime in oral squamous cell carcinoma (OSCC)

Das diversas neoplasias denominadas câncer de boca, mais de 95% tem o diagnóstico de carcinoma epidermóide (CE). O carcinoma epidermóide apresenta etiologia e patogênese complexas e não totalmente compreendidas determinando altos índices de morbidade e mortalidade. Por isso crescentes esforços têm sido empregados para a melhor compreensão dos eventos celulares, da susceptibilidade genética e dos fatores de risco que atuando em conjunto dão origem e atuam na progressão do carcinoma epidermóide de boca. Buscando novas facetas da carcinogênese oral e baseando-se no papel pró-carcinogênico que os hormônios esteróides desempenham nas neoplasias dos órgãos reprodutivos, alguns estudos passaram a avaliar o papel desses hormônios na patogênese e progressão do CE. O propósito deste trabalho foi avaliar a presença dos receptores de androgênio, estrogênio (alfa e beta) e da enzima aromatase em carcinomas epidermóides de boca (CE), através das técnicas de imuno-histoquímica, western-blotting e imunofluorescência, e comparar, com base no gênero (masculino e feminino), os resultados encontrados. Da amostra de CE utilizada, 30 eram pertencentes a pacientes do gênero feminino e 30 do masculino...

Oral squamous cell carcinoma (OSCC) is the main type of oral cancer presenting complex and not completely understood pathogenesis. OSCC implies quite significant mortality and morbidity rates and in spite of the vast amount of research and the advances in the field of oncology and surgery, the overall survival remains largely unchanged. Therefore, the identification of new pathways involved in the mechanisms of carcinogenesis can bring knowledge to the control and advent of new treatments. The role of androgens and estrogens in several endocrine-related malignancies is well known. The successful use of these hormones antagonists in the treatment of these neoplasms has prompted researches to investigate the presence of hormones receptors in other tissues and tumor types. The aim of this study was to asses through immunohistochemistry, western blot and immunoflorescence assays the expression, genderrelated, of androgen, estrogen alpha and beta receptors and aromatase enzyme in cases of OSSC. A total of 60 cases of OSCC (30 from males and 30 from females) were retrieved and submitted to immunohistochemical assay. Two OSCC cell lines were used for western blot and immunoflurescence techniques...

The interaction between aromatase, metalloproteinase 2,9 and cd44 in breast cancer / A interação entre aromatase, metalloproteinase 2, 9 e cd44 no câncer de mama

OBJECTIVE: This study intends to verify the expression levels and correlation of aromatase, matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) and CD44 in ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma (IDC) when both are found in the same breast. METHODS: One hundred and ten cases were evaluated by tissue microarray (TMA) and immunohistochemically screened with anti-aromatase polyclonal antibodies, anti-MMP-2 monoclonal antibodies, anti-MMP-9 policlonal antibodies and anti-CD44 monoclonal antibodies. RESULTS: Aromatase was expressed in IDC and DCIS in 63 (57.3 percent) and 60 (67 percent) of the cases respectively; MMP-2 was similarly expressed in IDC and DCIS in 15 (13.60 percent) cases; MMP-9 was positively expressed in IDC and DCIS in 83 (75.50 percent) and 82 (74.50 percent) cases, respectively; CD44 was positively expressed in IDC and DCIS in 49 (44.50 percent) and 48 (42.60 percent) of the cases, respectively; all of them were highly correlated (p<0,001). The correlation analysis found positive, statistically significant correlation, in IDC between aromatase and MMP-2 (p<0.001) and between aromatase and MMP-9 (p=0.034). Positive correlation between aromatase and MMP-2 (p<0.001) and between MMP-9 and CD44 (p=0.030) were found in DCIS. CONCLUSION: These results allow us to conclude that aromatase through local estrogen synthesis in breast tissue plays an important role in breast carcinogenesis, mainly influencing MMP-2 and MMP-9 which are important participants in tumor cell invasion and dependence of their connection to CD44 for action.

OBJETIVO: O objetivo desse estudo é verificar as expressões e correlações da aromatase, metalloproteinase 2 da matriz (MMP2), metalloproteinase 9 da matriz (MMP-9) e CD44 no carcinoma ductal in situ (CDIS) e carcinoma ductal infiltrativo (CDI) quando ambos estão presentes simultaneamente na mesma mama. MÉTODOS: Foram avaliados 110 casos pelo método de tissue microarray (TMA) e através da utilização de anticorpos policlonais antiaromatase, anticorpos monoclonais anti-MMP-2, anticorpos policlonais anti-MMP-9 e anticorpos monoclonais anti-CD44. RESULTADOS: A aromatase estava expressa de forma positiva no CDI e CDIS em 63 (57,3 por cento) e 60 (67 por cento) casos, respectivamente. A expressão de MMP-2 estava expressa de forma positiva em 15 (13,6 por cento) casos tanto no CDI, quanto no CDIS. A expressão da MMP-9 estava expressa de forma positiva em 83 (75,5 por cento) e 82 (74,5 por cento) casos de CDI e CDIS, respectivamente. A expressão de CD44 estava expressa de forma positiva em 49 (44,5 por cento) e 48 (42,6 por cento) casos de CDI e CDIS, respectivamente. Todos eles apresentando alta correlação (p<0,001). Na avaliação de correlação foi encontrada correlação positiva estatisticamente significante no CDI entre aromatase e MMP-2 (p<0,01) e entre aromatase e MMP-9 (p=0,034). Nos casos de CDIS houve correlação positiva estatisticamente significante entre aromatase e MMP-2 (p<0,001) e entre CD44 e MMP-9 (p=0,030). CONCLUSÃO: Após analisarmos os resultados de nosso estudo, podemos concluir que a aromatase, através da síntese de estrogênio local no tecido mamário, desempenha importante papel na carcinogênese mamária, principalmente influenciando a atuação da MMP-2 e da MMP-9, grandes responsáveis pela invasão celular tumoral que, por sua vez, provavelmente dependem de sua ligação a CD44 para poder desempenhar suas funções.

Estudio de análogos de la sulfonanilida que actúan como supresores de la expresión de la aromatasa / Study of sulfonanilide analogues acting as suppressants of the aromatase expression

Se realizó un estudio tridimensional cuantitativo de relación-estructura con nuevos análogos de la sulfonanilida que actúan como supresores de la expresión y actividad de la aromatasa en células de cáncer de mama, independientemente de la inhibición de la ciclooxigenasa-2 (COX-2). La superposición molecular de los ligandos en la plantilla fue llevada a cabo por el método Database Alignment. El mejor modelo estuvo constituido por la combinación de los campos estérico e hidrofóbico, los cuales arrojaron los siguientes parámetros: q² = 0,613 con siete componentes, R² no validado igual a 0,976, valor de F igual a 81,695; 0,347 y 0,047 para los errores estándar de predicción y estimación, respectivamente. Las contribuciones estérica e hidrofóbica fueron de 50,5 y 49,5% respectivamente. Los datos generados por el presente estudio podrían impulsar el diseño de nuevos y más potentes reguladores selectivos de la expresión de la aromatasa.

A three-dimensional quantitative structure-activity relationship study was performed with new Sulfonanilide analogue molecules acting as aromatase expression and activity suppressors in breast cancer cells independent of COX-2 inhibition. The molecular supression of the ligands in the grid was carried out by the DATA-BASE ALIGNMENT method. The best model formed by combining the esteric fields and hydrophobic fields yielded the following parameters: q² = 0.613 with seven components, not validated R² equal to 0.976, with an F value of 81.695, 0.347 and 0.047 for the standard errors of prediction and estimate, respectively. The contribution of esteric and hydrophobic fields was 50.5 and 49.5% respectively. data generated from this study may be used to design new and more potent selective aromatase expression regulators.

Regulação da massa corpórea pelo estrogênio e pela atividade física: [revisão] / Body mass regulation by estrogen and physical activity: [review]

A deficiência de esteroides gonadais femininos acelera o ganho de massa corpórea, mas os possíveis mecanismos centrais e periféricos envolvidos no aumento da ingestão alimentar e no ganho de massa adiposa que ocorrem nessa condição são pouco conhecidos. Em modelos animais, tanto a falta quanto os defeitos na ação do estrogênio causam aumento da massa corpórea, demonstrando claramente um possível papel desse esteroide no sobrepeso pós-menopausa. Sabe-se que a obesidade e o sobrepeso estão associados a diversas comorbidades que podem levar à morte prematura. Portanto, desvendar os mecanismos relacionados ao ganho de massa corpórea é de grande relevância, assim como desenvolver estratégias que possam prevenir o seu estabelecimento. A regulação do balanço energético está associada ao controle da massa corpórea, sendo o exercício físico um importante modulador desse parâmetro homeostático. Porém, a influência do exercício físico sobre o ganho de massa corpórea durante a deficiência de estrogênio é controversa e depende do protocolo de exercício utilizado. Neste estudo, pretendemos revisar os achados que relacionam a deficiência de estrogênio ao ganho de massa corpórea em animais e seres humanos.

Female steroid hormones deficiency leads to a significant increase in body mass, but the possible central and peripheral mechanisms involved in increased food ingestion and fat accumulation in this situation are still unknown. In animal models, the specific lack of estrogen or its action produce progressive body mass gain, clearly demonstrating the possible role of this hormone in overweight after menopause. Obesity and overweight correspond to a relevant human health problem that can lead to premature death. Therefore unraveling the mechanisms underlying body mass gain is of great relevance, as well as the development of strategies to prevent its establishment. Energy balance regulation is associated with the control of body mass, and physical exercise is an important modulator of this homeostatic parameter. However, the influence of physical exercise in mass gain development during estrogen deficiency is controversial and depends on the exercise protocol used. In this study, we intend to review the data on the effects of estrogen deficiency on body mass gain in humans and animal models.

Estrogênio e o sistema reprodutor masculino / Estrogen and male reproductive system

Objetivo: realizar uma revisão que demonstre a presença e a atuação do estrogênio sobre o sistema reprodutor masculino, propiciando um intercâmbio com o processo da infertilidade masculina. Material e métodos: A revisão foi baseada na pesquisa de artigos científicos no período relativo aos últimos 20 anos de produções científicas e publicações. Resultados: o desenvolvimento normal do sistema reprodutor masculino é fundamental nos mamíferos machos. Qualquer alteração nesse processo pode ocasionar um quadro de infertilidade. Acreditava-se que o desenvolvimento testicular e a manutenção da espermatogênese eram controlados, basicamente, pelas gonadotrofinas e pela testosterona. Contudo, evidências provenientes de diversas pesquisas com animais, incluindo relatos de casos em humanos, reforçaram a ideia de que o estrogênio tem papel importante na regulação do desenvolvimento e funcionamento testicular. O avanço das técnicas de Biologia molecular possibilitou a determinação da localização e ação do estrogênio, de seus receptores (ERa e ERb), assim como a determinação genética da enzima responsável pela conversão de androgênios em estrogênios, a aromatase. Conclusões: é possível relacionar a presença do estrogênio com a produção dos espermatozoides (número, maturação, motilidade e habilidade de fertilização), com a morfologia e fisiologia normal dos ductoseferentes e epidídimos e com a esteroidogênese das células testiculares. Contudo, há necessidade de mais estudos, muito embora já esteja evidente o envolvimento do estrogênio no processo reprodutivo masculino.

Objective: to make a review to demonstrate the presence and action of estrogen on male reproductive system, providing an exchange with the process of male infertility. Material and methods: the review was based on the research of papers published in a period covering the last 20 years of scientific productions and publications. Results: the normal development of male reproductive system is essential in male mammals. Any change in this process can cause infertility. It was believed that the testicular development and the maintenance of spermatogenesis were basically controlledby gonadotrophins and testosterone. However, evidence from studies with animals, including human case reports, reinforced the idea that estrogen has an important role in the regulation of testicular development and activity. The advancement of Molecular Biology techniques determined the location and action of estrogen, as well as its receptors (ERa and ERb), and allowed the genetic determination of the aromatase, enzyme responsible for conversion of androgens into estrogens. Conclusions: It is possible to relate the presence of estrogen to sperm production (number, maturation, motility, fertilization ability), to normal morphology and physiology of efferent and epididymis ducts, and to steroidogenesis in testicular cells. However, more studies are necessary, although the involvement of estrogen in male reproductive process be already apparent.

Expressão da aromatase em carcinomas de mama ductais invasivo e in situ presentes na mesma mama / Aromatase expression in invasive and in situ ductal carcinoma present in the same breast

OBJETIVO: Avaliar expressão da enzima aromatase nos carcinomas de mama ductais invasivos (CDI), in situ (CDIS), no epitélio e estromas adjacentes. MÉTODOS: Foram avaliados 45 espécimes cirúrgicos provenientes de mastectomias e quadrantectomias com CDI e CDIS concomitantes de pacientes com estadios clínicos I e II. A análise da expressão da enzima aromatase foi realizada por meio de anticorpos policlonais antiaromatase e categorização das amostras de acordo com intensidade e número de células coradas. RESULTADOS: Nos 45 casos de CDI a expressão da aromatase foi positiva em 32 espécimes (71 por cento) e negativa em 13 (29 por cento). Nos casos de CDIS, a positividade foi idêntica à observada no CDI, mostrando correlação positiva. No epitélio normal constatou-se expressão positiva em 19 casos (42,2 por cento) e negativa nos outros 26 (57,8 por cento), mostrando correlação positiva estatisticamente (p<0,01), quando comparada com CDI e CDIS. Na análise do estroma normal a expressão da aromatase foi observada em apenas sete (15,5 por cento) dos 45 casos avaliados, não apresentando correlação com nenhuma variável analisada para expressão da aromatase. A presença da aromatase no estroma tumoral foi positiva em 36 casos (80 por cento) e negativa em 9 (20 por cento), mostrando correlação estatisticamente com a expressão no CDI (p<0,01) e no CDIS (p<0,01). Ao se comparar a expressão da aromatase no CDI, CDIS, epitélio normal e estroma tumoral com os graus nuclear e histológico, tamanho tumoral e idade da paciente, não foram encontradas diferenças estatisticamente significantes. CONCLUSÃO: Os resultados revelaram alta correlação entre expressão da aromatase no CDI, CDIS, epitélio normal e estroma tumoral, sugerindo possível mecanismo de ação autócrina e parácrina desta enzima na gênese do câncer de mama.

OBJECTIVE: to evaluate the expression of aromatase in simultaneously invasive ductal carcinoma (IDC) and ductal carcinoma in situ (DCIS). METHODS: forty-five surgical samples were obtained from mastectomy and quadrantectomy with simultaneous IDC and DCIS of stage I and II patients. Aromatase was evaluated using antibodies anti-aromatase and the samples classified in accordance with the number and intensity of stained cells. RESULTS: Aromatase was expressed positively in 32(71 percent) and negatively in 13(29 percent) of the cases in the IDC. The same results were obtained in the DCIS showing a perfect positive correlation. In the normal epithelium,aromatase was positive in 19(42.2 percent) and negative in 26 (57.8 percent) and a positive correlation, statistically significant was obtained when compared with IDC and DCIS(p<0.01). Concerning the normal stroma, positivity was only 7 (15.5 percent) showing no correlation with aromatase expression. Aromatase was positive in 36(80 percent) of the tumor stroma and this result was statistically significant as in the IDC and DCIS. Comparing results of aromatase expression with nuclear grade, histological grade, tumor size and age no difference was found. CONCLUSION: our results demonstrated high correlation between aromatase expression in IDC, DCIS, normal epithelium and tumor stroma showing a possible autocrine and paracrine mechanism of this enzyme in breast cancer.

Aromatase gene and its effects on growth, reproductive and maternal ability traits in a multibreed sheep population from Brazil

We determined the polymorphism C242T of the aromatase gene (Cyp19) and its allelic frequency, as well as the effect of the variants on productive and reproductive traits in 71 purebred Santa Inês sheep, 13 purebred Brazilian Somali sheep, nine purebred Poll Dorset sheep, and 18 crossbred 1/2 Dorper sheep. The animals were genotyped using the PCR-RFLP technique. The influence of the animal's genotype on its performance or on the performance of its lambs was analyzed by the least square method. Another factor assessed was the importance of the animal's genotype in analysis models for quantitative breeding value estimates, and whether there were differences among the averages of breeding values of animals with different genotypes for this gene. In the sample studied, no AA individuals were observed; the AB and BB frequencies were 0.64 and 0.36, respectively. All Brazilian Somali sheep were of genotype BB. All 1/2 Dorper BB animals presented a lower age at first lambing, and the Santa Inês BB ewes presented a lower lambing interval. In these same genetic groups, AB ewes presented higher litter weight at weaning. This is evidence that BB ewes have a better reproductive performance phenotype, whereas AB ewes present a better maternal ability phenotype. However, in general, animals with genotype AB presented better average breeding values than those with genotype BB.

El splicing alternativo del exón 5 de la citocromo p450 aromatasa podría ser un mecanismo de regulación de la producción de estrógenos en humanos / Exon 5 alternative splicing of the cytochrome P450 aromatase could be a regulatory mechanism for estrogen production in humans

La enzima P450 aromatasa (P450Aro) participa en la síntesis de estrógenos a partir de andrógenos. La mutación c655G>A, descripta en forma heterocigota en una niña y en forma homocigota en un hombre adulto, ambos con déficit de aromatasa, genera la disrupción del sitio dador de splicing exón5-intrón5. Se ha postulado que la retención del intrón5 y la generación de una proteína truncada inactiva serían las consecuencias de esta mutación. Sorpresivamente, la paciente presentó desarrollo espontáneo de mamas y niveles puberales de estradiol, sugiriendo una actividad aromatasa (AA) residual. En principio postulamos que la mutación c655G>A generaría la pérdida del exón5 con conservación del marco de lectura, generándose una proteína con menor actividad que podría explicar el déficit parcial. La expresión del ARNm sin exón5 (ARNm- E5) en linfocitos de la paciente sugiere una asociación entre la pérdida del exón y la presencia de la mutación; posteriormente confirmada realizando ensayos de splicing en células Y1. Sin embargo, la expresión del cDNAE5 en células Y1 presentó una AA nula que no explicaría un déficit parcial. La expresión del ARNm-E5 fue detectada en placenta, testículo y adrenal humanos como una variante de splicing normal. Estos resultados indicarían la ocurrencia de splicing alternativo (SA) en la zona codificante de P450Aro como un posible mecanismo regulador de la producción de estrógenos en tejidos esteroidogénicos humanos. La mutación c655G>A podría alterar los mecanismos fisiológicos reguladores del SA del exón5 favoreciendo su exclusión. De esta forma, bajos niveles de ARNm+E5 podrían expresarse aun en presencia de la mutación explicando el fenotipo de déficit parcial observado en la paciente.

P450 aromatase (P450Aro), involved in androgen to estrogen conversion, is encoded by the CYP19 gene. P450Aro c655G>A mutation described in heterozygous form in a girl and in homozygous form in an adult male with P450Aro deficiency results in an aberrant splicing due to disruption of a donor splice site. A truncated inactive protein would be expected if intron5 is retained. Surprisingly, the girl described with this mutation showed spontaneous breast development and pubertal estradiol (E2) levels suggesting residual P450Aro activity (AA). Formerly, we postulate the in frame E5 skipping as a consequence of this mutation generating a protein with some degree of activity. When P450Aro mRNA expression was analysed from patient's lymphocytes, an aberrant spliced mRNA lacking E5 (-E5mRNA) was detected, suggesting an association between E5 skipping and the presence of the mutation. Splicing assays in Y1 cells confirmed this association. -Ex5 cDNA expression in Y1 cells resulted in an inactive protein that could not explain patient's phenotype. Exon 5 might be predicted as a poorly defined exon suggesting a susceptibility to splicing mutations and physiological alternative splicing (AS) events. Therefore, -Ex5mRNA was assessed as a natural occurring alternative transcript in normal human steroidogenic tissues. As P450Aro -E5mRNA expression was detected in human term placenta, prepubertal testis and prepubertal adrenal, we might speculate that AS of P450Aro coding region would occur in humans and would be involved in the complex AA regulation. Furthermore, tissue specific regulation of AS might suggest low expression of +E5mRNA from the c655G>A allele explaining residual AA evidenced in the affected girl.

Panorama actual de los inhibidores de la aromatasa en la inducción de la ovulación / Current view of aromatase inhibitors in ovulation induction

El panorama actual de la inducción de la ovulación en el tratamiento de la infertilidad está cambiando debido a la introducción reciente en este campo de los inhibidores de la aromatasa de tercera generación (IA). Estos agentes han mostrado su eficacia como inductores de la ovulación, mediante la inhibición de la aromatasa que convierte los andrógenos en estrógenos, necesarios para el desarrollo y la maduración folicular. Este mecanismo de acción hace a los IA ventajosos frente al citrato de clomifeno, por carecer de efectos antiestrogénicos sobre el endometrio y el moco cervical. Presentamos una revisión de los tópicos más importantes con base en estudios publicados acerca del uso clínico de los IA.

Estrogens and male reproduction: a new concept

The mammalian testis serves two main functions: production of spermatozoa and synthesis of steroids; among them estrogens are the end products obtained from the irreversible transformation of androgens by a microsomal enzymatic complex named aromatase. The aromatase is encoded by a single gene (cyp19) in humans which contains 18 exons, 9 of them being translated. In rats, the aromatase activity is mainly located in Sertoli cells of immature rats and then in Leydig cells of adult rats. We have demonstrated that germ cells represent an important source of estrogens: the amount of P450arom transcript is 3-fold higher in pachytene spermatocytes compared to gonocytes or round spermatids; conversely, aromatase activity is more intense in haploid cells. Male germ cells of mice, bank voles, bears, and monkeys express aromatase. In humans, we have shown the presence of a biologically active aromatase and of estrogen receptors (alpha and ß) in ejaculated spermatozoa and in immature germ cells in addition to Leydig cells. Moreover, we have demonstrated that the amount of P450arom transcripts is 30 percent lower in immotile than in motile spermatozoa. Alterations of spermatogenesis in terms of number and motility of spermatozoa have been described in men genetically deficient in aromatase. These last observations, together with our data showing a significant decrease of aromatase in immotile spermatozoa, suggest that aromatase could be involved in the acquisition of sperm motility. Thus, taking into account the widespread localization of aromatase and estrogen receptors in testicular cells, it is obvious that, besides gonadotrophins and androgens, estrogens produced locally should be considered to be physiologically relevant hormones involved in the regulation of spermatogenesis and spermiogenesis.