Estrogen receptor mRNA determined by Quantitative Real-Time PCR in the nasal mucosa of women taking oral contraceptives / Receptor ARNm de estrógeno determinado por PCR Cuantitativa en Tiempo Real en la mucosa nasal de mujeres tratadas con anticonceptivos orales

SUMMARY: Estrogen receptors (ER) have been identified in human nasal mucosa, but its physiologic and pathologic impacts are not totally established. ER have been demonstrated in nasal mucosa by several authors, mainly by immunohistochemical method in nasal mucosa samples surgically removed. The present study aimed to quantify ERα and ERβ mRNA concentration by using an absolute quantitative real-time PCR in cells from nasal mucosa smear of women under oral contraceptive therapy. Nasal epithelium smear samples were collected from 110 patients divided in two groups: 55 women who present regular menstrual cycle without using contraceptives and 55 women who present regular menstrual cycle and have been using oral contraceptives for more than 3 months. All the patients answered a rhinitis symptoms questionnaire. The current study showed the potential usefulness of nasal turbinate mucosa cell sourcing, collected through swab, for extracting useful RNA for gene expression. We have identified the predominant expression of ERα isoform in a ratio 10-15 times higher compared to ERβ isoform. There is a tendency for positive correlation between the ERb isoform and the rhinitis severity score.

RESUMEN: Se han identificado receptores de estrógeno (RE) en la mucosa nasal humana, sin embargo sus impactos fisiológicos y patológicos aún no están totalmente establecidos. Varios autores han demostrado RE en la mucosa nasal, principalmente por método inmunohistoquímico en muestras obtenidas quirúrgicamente. El presente estudio tuvo como objetivo cuantificar la concentración de ARNm de REa y REb mediante el uso de una PCR cuantitativa absoluta en tiempo real en células de frotis de mucosa nasal de mujeres bajo terapia anticonceptiva oral. Se recolectaron muestras de frotis de epitelio nasal de 110 pacientes divididas en dos grupos: 55 mujeres que presentan ciclo menstrual regular sin uso de anticonceptivos y 55 mujeres que presentan ciclo menstrual regular con uso de anticonceptivos orales durante más de 3 meses. Todas las pacientes respondieron un cuestionario de síntomas de rinitis. El estudio actual mostró la utilidad de la obtención de células de la mucosa de la concha nasal, recolectadas a través de un hisopo, para extraer ARN para la expresión génica. Hemos identificado la expresión predominante de la isoforma REμ en una proporción de 10 a 15 veces mayor en comparación con la isoforma REß. Hemos identificado la expresión predominante de la isoforma REα en una proporción de 10 a 15 veces mayor en comparación con la isoforma REß. Existe una tendencia a una correlación positiva entre la isoforma REß y la puntuación de gravedad de la rinitis.

Analysis of DFNA5 gene variant in a Chinese pedigree affected with late-onset non-syndromic hearing loss / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a Chinese pedigree affected with autosomal dominant late-onset non-syndromic hearing loss (NSHL).@*METHODS@#Clinical data of the pedigree were collected. Genomic DNA was extracted from peripheral blood samples of the proband and other family members. Trio whole exome sequencing was carried out for 19 396 genes to identify potential pathogenic variants. Sanger sequencing was carried out to verify the candidate variant in the pedigree.@*RESULTS@#The proband and his father were found to carry a c.1183+1delG p.? variant of the DFNA5 gene. The variant was confirmed to be co-segregating with the disease phenotype in the pedigree.@*CONCLUSION@#The c.1183+1delG p.? variant of the DFNA5 gene probably underlay the late onset NSHL in this pedigree. Above finding has enabled accurate genetic counseling for this pedigree.

Association of menopausal status, expression of progesterone receptor and ki67 to the clinical response to neoadjuvant chemotherapy in luminal breast cancer / Associação de status menopausal, expressão de receptor de progesterona e de Ki67 à resposta clínica à quimioterapia neoadjuvante no câncer luminal de mama

Abstract Objective To identify the biomarkers of response to neoadjuvant chemotherapy in early luminal breast cancer. Methods A cross-sectional study that included all patients with early or locallyadvanced luminal breast cancer submitted to neoadjuvant chemotherapy between 2013 and 2014. Demographic, clinic and pathologic data were retrieved from patient records. The expressions of the estrogen receptor (ER), the progesterone receptor (PR), and Ki67 were analyzed by immunohistochemistry (IHC). The status of the human epidermal growth factor receptor 2 (HER2) was evaluated by IHC and fluorescent in situ hybridization (FISH). Independent predictors of clinic and pathologic response were evaluated by stepwise logistic regression models and receiver operating characteristic (ROC) curve analysis. Results Out of 298 patients identified, 115 were included in the analysis. Clinical complete response (cCR) was observed in 43.4% of the patients (49/113), and pathologic complete response (pCR) was observed in 7.1% (8/115) of the patients. The independent predictors of cCR were premenopausal status (p < 0.001), low PR expression (≤ 50% versus > 50%; p = 0.048), and Ki67 expression ≥ 14% (versus < 14%; p = 0.01). Patients with cCR were more commonly submitted to breast conserving surgery (34.7% versus 7.8%; p < 0.001). Increasing cut-off points for Ki67 expression were associated with an increase in specificity and a decrease in sensitivity to identify patients with cCR. Conclusion Premenopausal status, lower PR expression and higher Ki67 expression were associated with a higher rate of cCR to neoadjuvant chemotherapy in luminal breast cancer.

Resumo Objetivo Identificar biomarcadores de resposta à quimioterapia neoadjuvante em câncer luminal de mama. Métodos Estudo transversal em que foram incluídas todas as pacientes com câncer luminal de mama em estádio inicial ou localmente avançado que foram submetidas a quimioterapia neoadjuvante nos anos de 2013 e 2014. Dados demográficos, clínicos e patológicos foram obtidos de prontuários médicos. As expressões de receptor de estrogênio (RE), de receptor de progesterona (RP), e de Ki67 foram avaliadas por imuno-histoquímica (IHQ). A expressão do receptor tipo 2 do fator de crescimento epidérmico humano (human epidermal growth factor receptor 2, HER2) foi avaliada por IHQ e hibridização in situ por imunofluorescência (HISI). Análises de regressão logística e de curva de característica de operação do receptor (COR) foram usadas para investigar fatores preditivos independentes de resposta clínica e patológica. Resultados De 298 pacientes identificadas, 115 foram incluídas na análise. Resposta clínica completa (RCc) foi observada em 43.4% das pacientes (49/113), e resposta patológica completa (RPc), em 7.1% (8/115). Os fatores preditivos independentes de RCc foram status menopausal (p < 0.001), baixa expressão de RP (≤ 50% versus > 50%; p = 0.048), e expressão de Ki67 ≥ 14% (versus < 14%; p = 0.01). Pacientes com RCc apresentaram maior probabilidade de serem submetidas a cirurgia conservadora da mama (34.7% versus 7.8%; p < 0.001). Aumento no ponto de corte para expressão de Ki67 foi associado a aumento da especificidade e redução da sensibilidade na identificação de pacientes com RCc. Conclusão Status premenopausal, baixa expressão de RP e maior expressão de Ki67 estiveram associados a maior taxa de RCc à quimioterapia neoadjuvante no câncer luminal de mama.

Estrogen signaling in the proliferative endometrium: implications in endometriosis / Sinalização pelo estrogênio no endométrio proliferativo: implicações na endometriose

SUMMARY Even though the physiological role of estrogen in the female reproductive cycle and endometrial proliferative phase is well established, the signaling pathways by which estrogen exerts its action in the endometrial tissue are still little known. In this regard, advancements in cell culture techniques and maintenance of endometrial cells in cultures enabled the discovery of new signaling mechanisms activated by estrogen in the normal endometrium and in endometriosis. This review aims to present the recent findings in the genomic and non-genomic estrogen signaling pathways in the proliferative human endometrium specifically associated with the pathogenesis and development of endometriosis.

RESUMO Embora esteja bem estabelecido o papel fisiológico do estrogênio no ciclo reprodutivo feminino e na fase proliferativa do endométrio, as vias de sinalização por meio das quais a ação do estrogênio é exercida no tecido endometrial são ainda pouco conhecidas. Nesse sentido, o avanço nas técnicas de cultura celular e a manutenção de células endometriais em cultivo possibilitaram a descoberta de novos mecanismos sinalizadores ativados pelo estrogênio no endométrio normal e na endometriose. Esta revisão tem o objetivo de apresentar as descobertas recentes envolvendo as vias de sinalização genômica e não genômica do estrogênio no endométrio proliferativo humano, especificamente associadas à patogênese e ao desenvolvimento da endometriose.

Expresión de cadherinas E y P en los tipos moleculares de cáncer de mama / Cadherins E and P expression in the molecular types of breast cancer.

La transición epitelial-mesenquimal es un proceso mediante el cual las células tumorales pierden sus marcadores epiteliales y facilita la migración a órganos distantes. En este proceso intervienen diversas proteínas de adhesión celular, tales como la cadherina E y la cadherina P. El presente estudio se realizó en 354 pacientes diagnosticadas de carcinoma ductal infiltrante de mama en seguimiento, en el Instituto de Oncología “Dr. Miguel Pérez Carreño” de Valencia, Venezuela. Se analizó la expresión de las dos moléculas por matrices de tejidos y se compararon los resultados obtenidos con las clases moleculares definidas por inmunohistoquímica, de acuerdo a la expresión de receptores de estrógeno (RE), receptores de progesterona (RP) y receptor del factor de crecimiento epidérmico humano 2 (HER2) y con la supervivencia global (SG). Con base a los resultados de RE, RP y HER2 se establecieron las clases moleculares, obteniendo los siguientes porcentajes: Luminal A 42,4%, Luminal B 20,3%, HER2 9% y triple negativo (TN) 28,2%. La expresión de cadherina E se observó conservada en la mayoría de los tumores de esta serie, 92,5% de los casos. Los tumores de fenotipo TN presentaron un porcentaje elevado (41,7%) con expresión ausente o reducida. La cadherina P se expresó en el 40,5% de los casos, y aunque expresada en todas las clases, la proporción fue significativamente mayor en los casos TN. No se apreció valor pronóstico significativo al analizar la SG a 5 años de las pacientes con tumores con ausencia o expresión reducida de cadherina E. La expresión de cadherina P presentó relación negativa con la SG.

The epithelial-mesenchymal transition is a process by which tumor cells lose their epithelial markers and migrate to distant organs. This process involves several cell adhesion proteins such as E-cadherin and P-cadherin. The present study was performed in 354 pacients diagnosed with breast infiltrating ductal carcinoma in the Oncology Institute “Dr. Miguel Pérez Carreño”, Valencia, Venezuela. The expression of 22 molecules was analyzed by tissue micro-arrays and the results were compared with the molecular clases established by immunohistochemistry, according to the expression of estrogen receptor (ER), progesterone (PR) and human epidermal growth factor receptor type 2 (HER2), and with the overall survival (OS). Based on the results of ER, PR and HER2 molecular classes according to the following percentages were established: Luminal A 42.4%, Luminal B 20.3%, 9% HER2 and 28.2% triple negative (TN). E-cadherin expression was observed conserved in most of the tumors of this series, 92.5% of cases. TN phenotype tumors showed a high percentage (41.7%) with absent or reduced expression. The P-cadherin was expressed in 40.5% of cases, although expressed in all classes; the proportion was significantly higher in cases TN. No significant prognostic value was observed when analyzing the overall five-year survival of patients with tumors with absent or reduced expression of E-cadherin. The P-cadherin expression had a negative relationship with the OS.

Gene expression of estrogen and oxytocin receptors in the uterus of pregnant and parturient bitches

In the canine species, the precise mechanisms of pregnancy maintenance and the initiation of parturition are not completely understood. The expression of genes encoding the receptors for estrogen (ERα mRNA) and oxytocin (OTR mRNA) was studied in the endometrium and myometrium during pregnancy and parturition in dogs. Real-time PCR was performed to quantify the levels of ERα mRNA and OTR mRNA in the uterus of bitches during early (up to 20 days of gestation), mid (20 to 40 days) and late pregnancy (41 to 60 days), and parturition (first stage of labor). All tissues expressed ERα and OTR mRNA, and are thus possibly able to respond to eventual estrogen and oxytocin hormonal stimuli. No statistically significant differences in the expression of ERα mRNA were verified in the endometrium and myometrium throughout pregnancy and parturition, but expression of OTR mRNA increased at both parturition and late pregnancy. We concluded that the increase of endometrial and myometrial OTR mRNA expression in dogs is not an event dependent on estrogenic stimulation. Moreover, the contractility response of the canine uterus to oxytocin begins during pregnancy and maintains myometrial activity. The expression of OTR mRNA in canine uterine tissues varied over time, which supports an interpretation that the sensitivity and response to hormone therapy varies during the course of pregnancy and labor. Further studies are needed to elucidate the factors underlying the synthesis of uterine oxytocin receptors and the possible role of ERβ rather than ERα in the uterine tissues during pregnancy and parturition in dogs.

Prognostic Significance of a Complete Response on Breast MRI in Patients Who Received Neoadjuvant Chemotherapy According to the Molecular Subtype

OBJECTIVE: To evaluate the relationship between response categories assessed by magnetic resonance imaging (MRI) or pathology and survival outcomes, and to determine whether there are prognostic differences among molecular subtypes. MATERIALS AND METHODS: We evaluated 174 patients with biopsy-confirmed invasive breast cancer who had undergone MRI before and after neoadjuvant chemotherapy, but before surgery. Pathology findings were classified as a pathologic complete response (pCR) or a non-pCR, and MRI findings were designated as a radiologic CR (rCR) or a non-rCR. We evaluated overall and subtype-specific associations between clinicopathological factors including the assessment categories and recurrence, using the Cox proportional hazards model. RESULTS: There were 41 recurrences (9 locoregional and 32 distant recurrences). There were statistically significant differences in recurrence outcomes between patients who achieved a radiologic or a pCR and patients who did not achieve a radiologic or a pCR (recurrence hazard ratio, 11.02; p = 0.018 and recurrence hazard ratio, 3.93; p = 0.022, respectively). Kaplan-Meier curves for recurrence-free survival showed that triple-negative breast cancer was the only subtype that showed significantly better outcomes in patients who achieved a CR compared to patients who did not achieve a CR by both radiologic and pathologic assessments (p = 0.004 and 0.001, respectively). A multivariate analysis found that patients who achieved a rCR and a pCR did not display significantly different recurrence outcomes (recurrence hazard ratio, 2.02; p = 0.505 and recurrence hazard ratio, 1.12; p = 0.869, respectively). CONCLUSION: Outcomes of patients who achieved a rCR were similar to those of patients who achieved a pCR. To evaluate survival difference according to molecular subtypes, a larger study is needed.

Effect of neoadjuvant chemotherapy on stromal CD10 antigens in breast cancer - A preliminary study.

Introduction: CD10 is a zinc-dependent peptidase (metalloproteinase). Stromal CD10 expression in breast cancer correlates with poor prognosis, oestrogen receptor negativity and higher grade. CD10 may be a potential target of new cancer therapies as it is involved in cleavage of doxorubicin. Aim: To evaluate the effect of neo-adjuvant anthracycline-based chemotherapy on status of stromal CD10 antigens in breast cancer. Materials and Methods: Patients with invasive breast cancer scheduled for anthracycline-based neo-adjuvant chemotherapy were included in the study. Tumor stromal CD10 expression was estimated before and after 3 cycles of chemotherapy, and change in its status was correlated with clinical response to chemotherapy. Results: 16 out of the 29 patients had strong CD10 expression; in these 16 patients, 14 (87.5%) were hormone receptor negative, and 14 (87.5%) had HER-2/neu overexpression. Stromal CD10 expression remained same in 13 out of 29 cases (44.83%) after chemotherapy. There was a change in CD10 expression in the remaining 16 cases (55.17%); in 13 cases (44.83%) it decreased from its pre-chemotherapy status, while its expression increased in 3 cases (10.34%). In cases of complete and partial clinical response, there was no increase in CD10 expression. Where CD10 expression had increased after chemotherapy, there was either a minor response or no response to chemotherapy. In 13 cases where CD10 expression had decreased, 12 cases had a clinical response to chemotherapy. Conclusions: Strong CD10 expression correlates with hormone receptor negativity and HER-2/neu overexpression. Stromal CD10 expression in breast cancer is not static and changes with neo-adjuvant anthracycline-based chemotherapy. A stable or decrease in CD10 expression correlates with complete or partial clinical response, while an increase in CD10 expression appears to correlate with poor clinical response. A larger series is required to determine the clinical significance of these changes. As stromal CD10 expression and its change with chemotherapy may have a prognostic significance, they should be documented in breast cancer patients before and after chemotherapy.

Prognostic Factors for Patients with Bone-Only Metastasis in Breast Cancer

PURPOSE: Bone is the most frequent site of metastasis among breast cancer patients. We investigated prognostic factors affecting survival following bone-only metastasis in breast cancer patients. MATERIALS AND METHODS: The medical records of breast cancer patients who were treated and followed at Gangnam Severance Hospital retrospectively reviewed to identify patients with bone-only metastasis. RESULTS: The median time from the diagnosis of bone-only metastasis to the last follow-up or death was 55.2 [95% confidence interval (CI), 38.6-71.9] months. The Kaplan-Meier overall survival estimate at 10 years for all patients was 34.9%. In the multivariate Cox regression model, bisphosphonate treatment [hazard ratio=0.18; 95% CI, 0.07-0.43], estrogen receptor positivity (hazard ratio=0.51; 95% CI, 0.28-0.94), and solitary bone metastasis (hazard ratio=0.32; 95% CI, 0.14-0.72) were significantly associated with longer overall survival in the bone-only recurrence group. Among the treatment modalities, only bisphosphonate treatment was identified as a significant prognostic factor. CONCLUSION: Identifying the factors influencing breast cancer mortality after bone-only metastasis will help clarify the clinical course and improve the treatment outcome for patients with breast cancer and bone-only metastasis. Bisphosphonates, as a significant prognostic factor, warrant further investigation.

Expresión de receptores de estrógenos y progesterona en el tracto genital de ovejas raza Texel / Expression of estrogen and progesterone receptors in the genital tract of Texel ewes

El comportamiento reproductivo de los ovinos varía entre las diferentes razas. Dentro de los factores que inciden en la capacidad reproductiva, el nivel de expresión de receptores de estrógenos y progesterona en su tracto genital parece tener un rol relevante. En el endometrio, oviducto y ovario los estrógenos y progesterona, regulan la expresión de numerosas proteínas comprometidas en su desarrollo morfofuncional. Factores genéticos como la raza estarian relacionados con los niveles de expresión de estos receptores. Las ovejas de raza Texel tienen elevados índices de fertilidad, son muy prolíficas y presentan un alto porcentaje de gestación múltiple lo cual podría tener relación con la expresión de estos receptores en el tracto reproductivo. El objetivo del presente estudio fue evaluar la expresión de receptores de estrógenos y progesterona en tracto genital de ovejas de raza Texel de alta prolificidad. La expresión de la proteína receptora de ambos receptores se detectó mediante análisis inmunohistoquímico y el nivel de expresión de los transcritos por RT-PCR en Tiempo Real Cuantitativo. Los resultados muestran expresión inmunohistoquímica del receptor de estrógeno principalmente en zonas glandulares y carunculares del endometrio. Se destaca además una menor expresión de ambos receptores en ovario, epitelio del oviducto y cervix. La expresión del receptor de progesterona a nivel inmunohistoquímico es bastante menor en cuanto a la señal destacándose marcas débiles en endometrio y ovarios. El nivel de expresión de los transcritos mantiene la misma distribución que las señales inmunohistoquimicas para ambos receptores. Concluimos que ambos receptores son expresados en el sistema reproductivo de ovejas de raza Texel en una distribución similar a lo encontrado en otras razas de ovejas quedando por definir si los niveles de expresión son similares en las distintas razas.

Reproductive performance of sheep varies between different races. Among the factors that affect the reproductive capacity, the level of expression of estrogen and progesterone receptors in the genital tract appears to have a relevant role. In the endometrium, oviduct and ovarian estrogen and progesterone regulate the expression of numerous proteins involved in morphofunctional development. Genetic factors such as race would be related to expression levels of these receptors. The Texel ewes have high fertility rates, are very prolific and have a high percentage of multiple births which could be related to the expression of these receptors in the reproductive tract. The aim of this study was to evaluate the expression of estrogen and progesterone receptors in the genital tract of ewes and Texel high prolificacy. The expression of the receptor protein of both receptors was detected by immunohistochemistry and the level of expression of the transcripts by Quantitative RT-PCR Real-Time. The results showed immunohistochemical expression of estrogen receptor mainly in areas of glandular and caruncular endometrium. It also highlights a lower expression of both receptors in ovarian tissue, epithelium of the oviduct and cervix. Expression of progesterone receptor immunohistochemical level is much lower with weaker signal in endometrium and ovaries. The expression level of transcripts remains the same distribution as immunohistochemical signals for both receptors. We conclude that both receptors are expressed in the reproductive system Texel ewes in a distribution similar to that found in other breeds of sheep, must be defined if the levels of expression are similar in different races.

Transcriptional regulation of bidirectional gene pairs by 17-β-estradiol in MCF-7 breast cancer cells

Using cDNA microarray analysis, we previously identified a set of differentially expressed genes in primary breast tumors based on the status of estrogen and progesterone receptors. In the present study, we performed an integrated computer-assisted and manual search of potential estrogen response element (ERE) binding sites in the promoter region of these genes to characterize their potential to be regulated by estrogen receptors (ER). Publicly available databases were used to annotate the position of these genes in the genome and to extract a 5’flanking region 2 kb upstream to 2 kb downstream of the transcription start site for transcription binding site analysis. The search for EREs and other binding sites was performed using several publicly available programs. Overall, approximately 40 percent of the genes analyzed were potentially able to be regulated by estrogen via ER. In addition, 17 percent of these genes are located very close to other genes organized in a head-to-head orientation with less than 1.0 kb between their transcript units, sharing a bidirectional promoter, and could be classified as bidirectional gene pairs. Using quantitative real-time PCR, we further investigated the effects of 17β-estradiol and antiestrogens on the expression of the bidirectional gene pairs in MCF-7 breast cancer cells. Our results showed that some of these gene pairs, such as TXNDC9/EIF5B, GALNS/TRAPPC2L, and SERINC1/PKIB, are modulated by 17β-estradiol via ER in MCF-7 breast cancer cells. Here, we also characterize the promoter region of potential ER-regulated genes and provide new information on the transcriptional regulation of bidirectional gene pairs.

Estudio de la expresión de los receptores de progesterona y estrógeno en el endometrio de ovejas alimentadas con dieta normal y dieta suplementada / Study of the expression of progesterone receptors in the endometrium of ewes with normal and supplementary diet

La influencia de la nutrición sobre la reproducción en ovinos es compleja y los resultados observados son variables. Planes nutricionales bajos pueden ser causa de una reducción de la sensibilidad del endometrio a la progesterona lo que afecta el desarrollo morfofuncional del endometrio en estadios tempranos de la preñez y por lo tanto va en directo detrimento de la sobrevida del embrión. La progesterona y los estrógenos actúan a través de receptores intracelulares específicos que median su efecto fisiológico. Por lo tanto el nivel de expresión de estos receptores es fundamental para el efecto hormonal. El objetivo del presente estudio fue analizar en forma conjunta la expresión de receptores de progesterona (RP) y estrógeno (RE), en el endometrio de ovejas en ciclo alimentadas con dieta suplementada y dieta normal. La expresión de la proteína receptora como del transcrito de ambos receptores se detectó mediante análisis inmunohistoquímico y RT-PCR en Tiempo Real. Los resultados muestran expresión inmunohistoquímica contra ambos receptores en zonas glándulares y carunculares, destacándose una intensa inmunorreacción en núcleo de células estromales y del epitelio glandular. Se detectó mayor expresión del transcrito de ambos receptores en ovejas alimentadas con suplemento respecto a las que no recibieron suplemento alimenticio.

The influence of nutrition on reproduction in sheep is complex and the reported results are variable. Low nutritional programs can cause a reduction in the sensitivity of the endometrium to progesterone which affects morphofunctional development of the endometrium in early stages of pregnancy and therefore is in direct expense of the survival of the embryo. Progesterone and estrogens act through specific intracellular receptors that mediate its physiological effect. Therefore the level of expression of these receptors is essential for the hormonal effect. The aim of this study was to analyze the expression of progesterone receptors (PR) and estrogen (ER) in the endometrium of sheep supplemented diet cycle and fed normal diet. The expression of the receptor protein and the transcript of both receptors was detected by immunohistochemistry and RT-PCR Real Time. The results show immunohistochemical expression against both receptors in caruncular and glandular areas, highlighting an intense immunoreaction in the nucleus of stromal cells and glandular epithelium. Were detected increased expression of the transcript of both receptors in sheep fed on the supplement that did not receive food supplement. We discuss the possible use of this information for applications in breeding programs in sheep breeding.

In vitro estrogenic activities of fenugreek Trigonella foenum graecum seeds.

Background & objectives: Trigonella foenum graecum commonly known as fenugreek, has been widely cultivated in Asia, Africa and Mediterranean countries for the edible and medicinal values of its seeds. Earlier reports show that fenugreek seeds provide a mastogenic effect resulting in enhanced breast size. However, very little is known about its estrogenic effect. The present study investigated the effect of chloroform extracts of fenugreek seeds (FCE) in breast cancer cells for its estrogenic effect, and to assess its capacity as an alternative to hormone replacement therapy (HRT). Methods: The effect of FCE on cell proliferation of estrogen receptor (ER) positive breast cancer cells, MCF-7 was studied by MTT assay at a concentration range of 20 to 320 μg/ml. The competitive ER binding assay (HAP assay) was done to find out the ER binding capacity of the extract. Transfection and reporter assay (DLR assay), and RT- PCR with an estrogen responsive gene pS2 were done to find out the transcriptional regulatory activity of FCE. Results: FCE stimulated the proliferation of MCF-7 cells, showed binding to ER (IC50 = 185.6 ± 32.8 μg/ml) and acted as an agonist for ER mediated transcription via ERE. It also induced the expression of estrogen responsive gene pS2 in MCF-7 cells. Interpretation & conclusion: Our study provided the evidence for estrogenic activities of fenugreek seeds. Further in vitro and in vivo studies could demonstrate its suitability as an alternative to HRT.

Influence of estradiol and triiodothyronine on breast cancer cell lines proliferation and expression of estrogen and thyroid hormone receptors / Efeito do estradiol e da triiodotironina na proliferação de linhagens celulares de câncer de mama e na expressão de receptores de estrógeno e hormônio tireoidiano

OBJECTIVE: To better understand the estrogen (E2) agonist action of triiodothyronine (T3) the effects of these hormones on ER negative MDA-MB-231 breast cancer cells were compared with those on S30, a clone of MDA-MB-231 stably transfected with ERα cDNA, in terms of proliferation and modulation of hormone receptors. RESULTS: Growth experiments showed that MDA-MB-231 was not modulated by any hormone or tamoxifen (TAM). Treatment with E2, 10-8M or 10-9M had little effect on S30 proliferation. T3 at 10-8M significantly inhibited proliferation. This effect was not reverted by TAM. Treatments with 10-8M concentration of E2 or T3 reduced ERα gene expression in S30, an effect partially blocked by association with TAM, with no effect on TR expression. These results suggest that, in S30, 10-8M T3 has a similar action to E2 relative to ERα gene modulation. CONCLUSIONS: Such results emphasize the need of determining T3 levels, before the introduction of antiestrogenic forms of treatment in breast cancer patients.

OBJETIVO: Para compreender melhor a ação da triiodotironina (T3) agonista de estrógeno (E2), foram comparados os efeitos destes hormônios em células de câncer de mama MDA-MB-231 ER negativas com um clone de MDA-MB-231, transfectado estavelmente com o cDNA de ERα (S30), em termos de proliferação e modulação dos receptores hormonais. RESULTADOS: Experimentos de crescimento mostraram que MDA-MB-231 não foi modulada por qualquer hormônio ou pelo tamoxifeno (TAM). O crescimento de S30 foi essencialmente inalterado por tratamento com E2 10-9M ou 10-8M, mas T3 10-8M inibiu significativamente a proliferação quando comparada a ambas concentrações de E2. Esse efeito não foi revertido pelo TAM, sugerindo um resultado não genômico, independente de ERE. Tratamentos com 10-8M de E2 ou de T3 reduziram a expressão do gene ERα em S30, efeito parcialmente impedido pela associação com TAM, sem efeito na expressão de TR. Os resultados sugerem que, em S30, T3 10-8M tem ação semelhante ao E2 com relação à modulação do gene ERα. CONCLUSÕES: Esses resultados enfatizam a necessidade de dosagem de T3 circulante antes da introdução do tratamento antiestrogênico no câncer de mama.

Expression of basal and luminal cytokeratins in breast cancer and their correlation with clinicopathological prognostic variables.

Background : Normal breast ducts contain at least 3 types of epithelial cells: luminal (glandular) cells, basal/myoepithelial cells and stem cells. Myoepithelial and luminal epithelia can be distinguished by their different cytokeratin expression patterns. The aim of this study is to evaluate the expression of some prognostic biomarkers (ER, PR and HER2), as well as histological grading and lymph node status in cytokeratin-based groups of breast cancer. Objective: To evaluate the correlation between expression of basal and luminal markers and hormonal receptors, HER2/neu, age, grade and lymph node status in breast-invasive ductal carcinoma. Materials and Methods : Sixty-seven formalin-fixed and paraffin-embedded breast cancer specimens (of invasive ductal carcinoma, 'NOS' type) which had already been studied for ER, PR and HER2/neu were selected. Data concerning age, tumor grade and lymph node status were also obtained from archives. Expression of basal (CK5/6) and luminal (CK7) cytokeratins was detected by immunohistochemistry. Stained sections were classified according to the intensity of staining and the percentage of stained cells. Results : We categorized the cases into 3 distinct phenotype groups: pure luminal, basal phenotype and null. Pure basal, mixed basal and luminal groups were classified as expressing a basal phenotype. There was a significant difference in the ER and/or PR expression between those 3 groups and a significant association between ER and/or PR negativity and basal phenotype expression. There was no significant difference in HER2/neu expression, age of the patients, tumor grade and lymph node status between the 3 cytokeratin-based groups and no significant association between lymph node status and basal phenotype expression. Conclusion : We found that to gain a real association between basal phenotype and prognostic markers, we should use a cocktail or a panel of different biomarkers to correctly determine basal-like phenotype of breast cancers. This approach guarantees more concordance with gene expression-based studies.

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.

Association of CYP1A1*2 polymorphisms with breast cancer risk: a case control study.

BACKGROUND: The Cytochrome P-4501A1 (CYP1A1) gene, located on chromosome 15q, is involved in the metabolism of carcinogens mainly polycyclic aromatic hydrocarbons as well as estrogen. It is considered as candidate gene for low-penetrance breast cancer susceptibility. Hence the present study aims to discuss the role of CYP1A1 polymorphisms in breast cancer. MATERIALS AND METHODS: A total of 250 breast cancer patients and the same number of healthy age-matched controls were analyzed for the polymorphism of CYP1A1*2 by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: In the present study, association of CYP1A1*2 (Ile 462Val) polymorphism with breast cancer was studied. Only one breast cancer patient was observed to be homozygous for Val allele but none among controls. The frequency of heterozygous Ile/Val genotype was found to be increased significantly in breast cancer patients (68.1%) as compared to controls (51.0%). Higher frequency of heterozygotes for Val allele was observed among premenopausal breast cancer patients and patients with high BMI, positive for HER2/neu status and advanced stage of the disease in comparison to the corresponding groups. No significant association of CYP1A1*2 polymorphism was observed with occupation, estrogen receptor and progesterone receptor status of breast cancer patients. CONCLUSIONS: In conclusion, our results suggest a significant correlation between CYP1A1*2 expression and the occurrence of breast cancer.

Expression of estrogen and progesterone receptor genes in endometrium, myometrium and vagina of postmenopausal women treated with estriol / Expresión de los genes receptores de estrógeno y progesterona en el endometrio, miometrio y vagina en mujeres posmenopáusicas tratadas con estriol

CONTEXT AND OBJECTIVE: Estriol is an estrogen with considerably weaker stimulatory effects on endometrial proliferation than estradiol. A study was conducted to determine the level of estrogen receptors (ERs) and progesterone receptors (PRs) in women who received 14-day vaginal estriol therapy, compared with those who did not receive this therapy. ER and PR gene expression was analyzed in the endometrium, myometrium and vagina of postmenopausal women treated with estriol. DESIGN AND SETTING: Analytical cross-sectional study, at the Research Institute of the Polish Mothers' Memorial Hospital, Lodz, Poland. METHODS: Twenty-seven postmenopausal women (57-74 years of age) were included in the study. All of them were waiting for per vaginam hysterectomy or plastic surgery on the vagina and perineum because of uterine prolapse. ER and PR gene expression was determined by means of the technique of reverse transcription polymerase chain reaction (RT-PCR). RESULTS: In the estriol-treated patients, in comparison with the control group, a significant increase in ER gene expression was observed in the endometrium and vagina, while enhanced PR gene expression was found in the endometrium. However, under histological examination of the endometrium, estrogen stimulation of low and medium degree was diagnosed for 21.4 percent and 14.3 percent of the estriol-treated women, respectively. CONCLUSION: The results obtained suggest that the women who received 14 days of treatment with vaginal estriol had higher ER and PR mRNA levels. No difference between these groups regarding endometrial proliferation was observed.

CONTEXTO Y OBJETIVO: El estriol es un estrógeno con un efecto estimulatorio bastante más débil sobre la proliferación endometrial que el estradiol. Se realizó un estudio para determinar los efectos de una terapia vaginal de 14 días con estriol, sobre el nivel de receptores de estrógeno (ER) y receptores de progesterona (PR), comparado con mujeres sin esa terapia. La expresión de los genes de ER y PR se analizó en el endometrio, miometrio y vagina de mujeres posmenopáusicas tratadas con estriol. DISEÑO Y UBICACIÓN: Estudio Transversal analítico, en el Instituto de Investigación del Hospital de la Madre Polaca en Lodz, Polonia. MÉTODOS: Se incluyeron veintisiete mujeres posmenopáusicas (de 57 a 74 años) en el estudio. Todas ellas estaban en espera de una histerectomía per vaginam o de cirugía plástica de la vagina y del perineo debido a un prolapso del útero. La expresión de genes de los receptores ER y PR se estableció por la técnica de RT-PCR. RESULTS: En las pacientes tratadas con estriol en comparación con el grupo de control, se observó un aumento significativo de la expresión del gen de ER en el endometrio y la vagina, mientras que un aumento de la expresión del gen de PR se encontró en el endometrio. De todas formas, en el examen histológico del endometrio, se diagnosticó estimulación estrogénica de bajo y medio grado en el 21.4 por ciento y en el 14.3 por ciento de las mujeres tratadas con estriol, respectivamente. CONCLUSIONES: Los resultados obtenidos sugieren que un tratamiento de 14 días con estriol en pacientes, aunque aumentan el nivel de ER y de PR mRNA, tiene muy poco o ningún efecto sobre la proliferación endometrial.

Impact of Grade, Hormone Receptor, and HER-2 Status in Women with Breast Cancer on Response to Specific Chemotherapeutic Agents by in vitro Adenosine Triphosphate-based Chemotherapy Response Assay

This study was designed to assess whether histological and biological factors of breast cancer can predict chemoresponse to specific agents. Adenosine triphosphate-based chemotherapy response assay (ATP-CRA) was employed to retrieve chemoresponse to 5-fluorouracil (5-FU), doxetaxel, doxorubicin, epirubicin, and paclitaxel in 49 patients. Tumors with high histologic and nuclear grade have higher response rate to doxorubicin (P<0.05) and palitaxel (P<0.05). Estrogen receptor (ER)-negative tumors respond well to doxorubicin (P=0.038), and progesterone receptor (PR)-negative tumors to 5-FU (P=0.039), doxetaxel (P=0.038), doxorubicin (P=0.000), epirubicin (P=0.010), and paclitaxel (P=0.003). Among the breast cancer subtypes determined by ER, PR, and HER-2 immunohistochemical stains, the HER-2+/ER- subtype has a higher response rate to doxorubicin (P=0.008). This in vitro result suggests that the combination of histologic and nuclear grade, hormone receptor, and HER-2 status can be a predictive factor of response to specific chemotherapy agents. Further in vivo study should be followed for clinical trials.

Clasificación molecular del cáncer de mama / Molecular classification of breast cancer

Breast cancer is classified based on clinical stage, cellular morphology and immunohistochemical analysis. More precise prognostic factors are necessary to aid with therapeutic decisions. Breast cancer subtypes that differ in their genetic expression and prognosis have been determined using cDNA microarrays. These findings confirm the differences between the phenotypes and provide new knowledge about the biology of breast cancer. Based on the presence or absence of expression of the estrogen receptor (ER), breast cancer is divided in two groups: ER+ and ER-. Genetic expression profile has identified two subtypes of the ER+ tumors: luminal A and luminal B. ER- tumors also include two subtypes, the HER2+ and the basal type. These subtypes differ in their biology and both demonstrate short disease-free periods after treatment and poorer outcome. This classification has shown the relationship between cDNA microarrays and clinical outcome of these tumors. This classification is proposed as a method of identifying those patients who will demonstrate better results with the different adjuvant modalities.