Role of steroid receptor-associated and regulated protein in tumor progression and progesterone receptor signaling in endometrial cancer / 中华医学杂志(英文版)

BACKGROUND@#Steroid receptor-associated and regulated protein (SRARP) suppresses tumor progression and modulates steroid receptor signaling by interacting with estrogen receptors and androgen receptors in breast cancer. In endometrial cancer (EC), progesterone receptor (PR) signaling is crucial for responsiveness to progestin therapy. The aim of this study was to investigate the role of SRARP in tumor progression and PR signaling in EC.@*METHODS@#Ribonucleic acid sequencing data from the Cancer Genome Atlas, Clinical Proteomic Tumor Analysis Consortium, and Gene Expression Omnibus were used to analyze the clinical significance of SRARP and its correlation with PR expression in EC. The correlation between SRARP and PR expression was validated in EC samples obtained from Peking University People's Hospital. SRARP function was investigated by lentivirus-mediated overexpression in Ishikawa and HEC-50B cells. Cell Counting Kit-8 assays, cell cycle analyses, wound healing assays, and Transwell assays were used to evaluate cell proliferation, migration, and invasion. Western blotting and quantitative real-time polymerase chain reaction were used to evaluate gene expression. The effects of SRARP on the regulation of PR signaling were determined by co-immunoprecipitation, PR response element (PRE) luciferase reporter assay, and PR downstream gene detection.@*RESULTS@#Higher SRARP expression was significantly associated with better overall survival and disease-free survival and less aggressive EC types. SRARP overexpression suppressed growth, migration, and invasion in EC cells, increased E-cadherin expression, and decreased N-cadherin and Wnt family member 7A ( WNT7A ) expression. SRARP expression was positively correlated with PR expression in EC tissues. In SRARP -overexpressing cells, PR isoform B (PRB) was upregulated and SRARP bound to PRB. Significant increases in PRE-based luciferase activity and expression levels of PR target genes were observed in response to medroxyprogesterone acetate.@*CONCLUSIONS@#This study illustrates that SRARP exerts a tumor-suppressive effect by inhibiting the epithelial-mesenchymal transition via Wnt signaling in EC. In addition, SRARP positively modulates PR expression and interacts with PR to regulate PR downstream target genes.

Pólipos endometriais e seu risco de malignização: aspectos epidemiológicos, clínicos e imunoistoquímicos / Endometrial polyps and the risk of malignancy: epidemiological, clinical and immunohistochemical aspects

Pólipos endometriais são neoformações resultantes de uma hiperplasia focal da camada basal do endométrio associada a um hiperestímulo hormonal. Sua etiologia ainda não está bem estabelecida, não havendo consenso sobre sua história natural, seu real significado como entidade patológica e sua relação com a neoplasia endometrial. Os pólipos endometriais são a principal indicação de histeroscopia cirúrgica, sem que haja, no entanto, um protocolo definido para seu melhor manejo. Uma visão abrangente sobre essa condição pode auxiliar na escolha da conduta mais adequada. (AU)

Endometrial polyps are neoformations that result from focal hyperplasia of the endometrial basal layer associated with hormonal hyperstimulation. Their pathogenesis is still unclear, and there is no consensus on their natural history, actual relevance as pathologic entities, and relationship with endometrial neoplasia. Endometrial polyps are the most frequent indication of surgical hysteroscopy, but their optimal management remains controversial. Therefore, an overview of this condition may help choosing the most adequate treatment strategies.(AU)

Claudina-3 e Claudina-4, potenciais marcadores de agressividade no carcinoma endometrial Tipo I / Claudin -3 and Claudin-4 potential aggressiveness markers in endometrial carcinoma Type I

O Carcinoma de Endométrio é a neoplasia epitelial maligna que acomete mulheres no pré, pós e peri -menopausa. Segundo diferenças endócrinas, metabólicas, fatores de risco e morfologia é classificado em dois grandes grupos: Tipo I (endometrióide) e Tipo II (não endometrióide). O Tipo I perfaz 90 % dos carcinomas de endométrio e sua patogenia esta ligada a exposição excessiva ao estrógeno. O Tipo II é incomum, com fatores predisponentes menos conhecidos. O sintoma mais relevante do carcinoma de endométrio é o sangramento pós-menopausa. Seu diagnóstico é feito pela comprovação histológica, associada aos exames de imagem e laboratoriais. Seu tratamento é fundamentalmente cirúrgico. O fator prognóstico mais importante é a presença ou ausência de metástase nos linfonodos regionais. Atualmente, buscam-se marcadores biológicos e teciduais que indiquem pior prognóstico. Este trabalho verificou a imunoexpressão da claudina-3 (CLDN3) e claudina-4 (CLDN4) nos carcinomas de endométrio Tipo I e Tipo II, relacionando-as com endométrio proliferativo e atrófico, aspectos clínicos, anatomopatológicos, perfil hormonal, índice de proliferação celular e expressão da p53, na tentativa de estabelecer a importância destas proteínas na progressão e agressividade tumoral e o seu valor prognóstico. Foram estudados 79 casos de carcinoma de endométrio e comparados com 74 endométrios normais. Avaliou-se a imunoexpressão das CLDNs 3 e 4, receptor estrogênico, receptor de progesterona, índice de proliferação celular (Ki67) e p53, pela técnica de imunoistoquímica. Observou-se que o padrão de coloração da membrana para CLDN3 se mostrou difuso nos carcinomas, quando comparado com os endométrios normais que exibiu padrão focal. O número de células marcadas com CLDN3 estava diminuído nos carcinomas Tipo I, porém com intensidade aumentada...

Carcinoma of the Endometrium is a malignant epithelial tumor which affects pre, peri and post-menopausal women. It is classified into two major groups, according to endocrine, metabolic risk factors and morphological differences: Type I (endometrioid) and Type II (non-endometrioid). Type I accounts for 90% of all endometrial carcinomas and its pathogenesis is linked to excessive estrogen exposure. The Type II is less common, with poorly defined predisposing factors. The most important symptom of endometrial carcinoma is postmenopausal bleeding. Diagnosis is achieved through histological evidence, in association with imaging and laboratorial exams. Treatment is primarily surgical. The most important prognostic factor is the presence or absence of metastases in regional lymph nodes. Bio and tissue markers that indicate worse prognosis are the focus of current research. This study examined the immunoexpression of claudin-3 (CLDN3) and claudin-4 (CLDN4) in endometrial carcinomas Type I and Type II, and their relation to proliferative and atrophic endometrium, clinical and pathological features, hormonal status, proliferation index and p53 expression, in an attempt to establish the importance of these proteins in tumor progression and aggressiveness and their prognostic value. Seventy-nine cases of endometrial carcinoma were studied and compared with 74 normal endometria. The immunoexpression of CLDNs 3 and 4, estrogen receptor, progesterone receptor, cell proliferation index (Ki67) and p53 were all evaluated by immunohistochemistry. Observation verified that the pattern of membrane staining for CLDN3 was diffuse in carcinomas compared with normal endometrium which presented a focal pattern. The number of cells stained with CLDN3 was lower in Type I carcinomas, while staining intensity was greater...

The Synergistic Apoptotic Interaction of Indole-3-Carbinol and Genistein with TRAIL on Endometrial Cancer Cells

Induction of apoptosis in target cells is a key mechanism by which chemotherapy promotes cell killing. The purpose of this study was to determine whether Indole-3-Carbinol (I3C) and Genistein in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induce apoptosis in endometrial cancer cell (Ishikawa) and to assess apoptotic mechanism. The MTT assay and flow cytometry were performed to determine cell viability and cell cycle. The induction of apoptosis was measured by caspase-3 activity test, DNA fragmentation assay, annexin V binding assay and western blot analysis. There was no effect in cell growth inhibition and cell cycle progression alone or in two-combination. However, the treatment of I3C and Genistein followed by TRAIL showed significant cell death and marked increase in sub-G1 arrest. Three-combination treatment revealed elevated expression of DR4, DR5 and cleaved forms of caspase-3, caspase-8, PARP. The Flip was found down regulated. Moreover, increase in caspase-3 activity and DNA fragmentation indicated the induction of apoptosis. The results indicate that I3C and Genistein with TRAIL synergistically induced apoptosis via death receptor dependent pathway. Our findings might provide a new insight into the development of novel combination therapies against endometrial cancer.

Canonical and noncanonical Wnt pathways: a comparison between endometrial cancer type I and atrophic endometrium in Brazil / Vias Wnt canônica e não canônica: uma comparação entre o câncer endometrial tipo I e endométrio atrófico no Brasil

CONTEXT AND OBJECTIVE: The Wnt pathway is involved in tumorigenesis of several tissues. For this reason, we proposed to evaluate Wnt gene expression in endometrial cancer type I. DESIGN AND SETTING: Cross-sectional study on materials gathered from the tissue bank of the Department of Pathology, Universidade Federal de São Paulo. METHODS: Endometrial specimens were obtained from surgeries performed between 1995 and 2005 at São Paulo Hospital, Universidade Federal de São Paulo. The material was divided into two groups according to tissue type: Group A, atrophic endometrium (n = 15); and Group B, endometrial adenocarcinoma (n = 45). We compared the immunohistochemical expression of Wnt1, Frizzled-1 (FZD1), Wnt5a, Frizzled-5 (FZD5) and beta-catenin between endometrial cancer type I and atrophic endometrium. RESULTS: Regarding Wnt1, FZD1 and Wnt5a expression, no significant association was observed between the groups. A significant association was observed between the groups in relation to FZD5 expression (P = 0.001). The proportion of FZD5-positive samples was significantly higher in group A (80.0 percent) than in group B (31.1 percent). Regarding the survival curve for FZD5 in group B, we did not find any significant association between atrophic endometrium and endometrial adenocarcinoma. We also did not find any significant association regarding beta-catenin expression (P = 1.000). CONCLUSION: FZD5 is downregulated in endometrial adenocarcinoma, in comparison with atrophic endometrium.

CONTEXTO E OBJETIVO: A via Wnt está envolvida na tumorigênese de diversos tipos de tecidos. Por essa razão, propusemo-nos a avaliar a expressão de genes da família Wnt no câncer endometrial tipo I. TIPO DE ESTUDO E LOCAL: Estudo transversal com coleta de materiais do banco de tecidos do Departamento de Patologia da Universidade Federal de São Paulo. MÉTODOS: Amostras endometriais foram obtidas de cirurgias que ocorreram entre 1995 e 2005 no Hospital São Paulo, Universidade Federal de São Paulo. Foram separados dois grupos segundo o tipo de tecido obtido: grupo A, com endométrio atrófico (n = 15); e grupo B, com adenocarcinoma endometrial (n = 45). Comparamos a expressão imunoistoquímica de Wnt 1, Frizzled-1 (FZD1), Wnt 5a, Frizzled-5 (FZD 5) e beta-catenina entre câncer endometrial tipo I e endométrio atrófico. RESULTADOS: Na expressão do Wnt1, FZD1 e Wnt5a, não observamos associação significante entre os grupos. Na expressão do FZD5, encontramos associação significante entre os grupos (P = 0,001). A proporção de positividade do FZD5 foi significantemente maior no grupo A comparado ao grupo B (31,1 por cento). Em relação à curva de sobrevida para o FZD5 no grupo B, não tivemos associação significante entre endométrio atrófico e adenocarcinoma do endométrio. Também não observamos associação significante na expressão da beta-catenina (P = 1,000). CONCLUSÃO: FZD5 é downregulated no adenocarcinoma endometrial quando comparado ao endométrio atrófico.

Efeitos do tamoxifeno sobre a expressão das proteínas TGF-β e p27 em pólipos e endométrio adjacente de mulheres após a menopausa / Effects of tamoxifen on the expression of TGF-β and p27 proteins in polyps and adjacent endometrium in postmenopausal women

OBJETIVO:avaliar os efeitos do uso do tamoxifeno sobre a expressão das proteínas TGF-β e p27 em pólipos e endométrio adjacente de mulheres após a menopausa. MÉTODOS: estudo prospectivo, com 30 mulheres, após a menopausa com diagnóstico de câncer de mama, usuárias de tamoxifeno (20 mg/dia), que apresentavam diagnóstico de pólipo endometrial suspeito por meio de ultrassonografia transvaginal, submetidas à histeroscopia diagnóstica e cirúrgica para retirada dos pólipos e do endométrio adjacente. Realizado estudo imunoistoquímico para verificar a expressão das proteínas TGF-β e p27 nos pólipos e no endométrio adjacente. A quantificação dessas proteínas foi realizada por morfometria. RESULTADOS: a média de idade foi 61,7 anos; média da idade na menopausa, 49,5 anos; e o tempo médio de uso do tamoxifeno, de 25,3 meses. A concentração média de células positivas para proteína TGF-β no pólipo epitélio glandular e estroma foi 62,6±4,5 células/mm². Para a p27, no pólipo epitélio glandular, foi de 24,2±18,6 cel/mm² e estroma 19,2±15,2 cel/mm². Não houve diferença significante entre a expressão do TGF-β e p27 nos epitélios glandulares dos pólipos e do endométrio adjacente. A expressão das proteínas do pólipo e endométrio adjacente com os seus respectivos epitélios glandular e estromal apresentou diferença significativa para a proteína p27 (r=0,9, p<0,05). CONCLUSÕES: concluímos que a expressão do TGF-β não está relacionada ao efeito do tamoxifeno sobre o crescimento dos pólipos endometriais, mas a ausência de malignização dos pólipos induzida pelo tamoxifeno pode ser explicada pela alta expressão da proteína p27 no seu epitélio glandular.

PURPOSE: to evaluate the effects of tamoxifen on the expression of TGF-β and p27 proteins in polyps and adjacent endometrium of women after menopause. METHODS: prospective study with 30 post-menopausal women with diagnosis of breast cancer, taking tamoxifen (20 mg/day), presenting diagnosis of suspect endometrial polyps through transvaginal ultrasonography, and submitted to diagnostic and surgical hysterectomy to withdraw the polyps and adjacent endometrium. A immunohistochemical study has been done to verify the expression of the TGF-β and p27 proteins in the polyps and adjacent endometrium. These proteins' quantification has been done by morphometry. RESULTS: the patients' average age was 61.7 years old; their average age at the menopause onset was 49.5; and the average of using tamoxifen was 25.3 months. The average concentration of positive cells for TGF-β protein in the glandular and stroma polyp epithelium was 62.6±4.5 cells/mm². For the p27, in the glandular polyp epithelium, it was 24.2±18.6 cells/mm² and for the stroma, 19.2±15.2 cells/mm². There was no significant difference between the expression of TGF-β and p27 in the glandular epithelial form the polyps and the adjacent endometrium. The expression of proteins in the polyp and adjacent endometrium with its respective glandular and stroma epithelium showed a significant difference for the p27 protein (r=0.9, p<0.05). CONCLUSIONS: we have concluded that the TGF-β expression is not related to the effect of tamoxifen on the growing of endometrial polyps, but the absence of polyps' malignization by tamoxifen may be explained by the high expression of p27 protein in its glandular epithelium.

Clinial implication of tThe expression of Aurora B in normal endometrium and endometrial carcinoma / 华中科技大学学报（医学）（英德文版）

The expression of Aurora B in normal endometria and endometrial carcinomas and its relation with clinicopathologic parameters of endometrial carcinomas were investigated. Streptavidin-biotin peroxidase (SP) immunohistochemical technique was used to detect the expression of Aurora B in 10 cases of normal proliferative phase endometria, 10 cases of normal secretory phase endometria and 72 cases of endometrial carcinomas respectively. According to the 1988 International Federation of Gynecology and Obstetrics (FIGO) grade, there were 37 patients in grade 1, 23 in grade 2 and 12 in grade 3 respectively. According to the FIGO stage, there were 59 patients in stage I-II and 13 patients in stage III-IV. Aurora B was expressed in both normal proliferative phase endometria, secretory phase endometria and endometrial carcinomas, but its positive labeling index (PLI) in proliferative phase endometria was significantly higher than that in secretory phase endometria (P1/2 myometrial invasion (all P<0.01). Aurora B exerts its functions in the replication of normal endometrial glandular cells; Expression of Aurora B is significantly correlated with biologic behavior of endometrial carcinoma, indicating that Aurora B may be a promising prognostic factor in endometrial carcinoma.

Immunohistochemical Analysis of CD44s and CD44v6 in Endometriosis and Adenomyosis: Comparison with normal, hyperplastic, and malignant endometrium

The expression patterns of CD44s and CD44v6 were immunohistochemically compared with those of normal, hyperplastic and malignant endometrium. In normal endometria (n=37), endometrioses (n=46) and adenomyoses (n=20), the surface and glandular epithelial cells were negative for CD44s and CD44v6 in a proliferative pattern and positive in a secretory pattern, whereas the stroma was only positive for CD44s in both proliferative and secretory patterns. The endometrial hyperplasia (4 simple and 9 complex) had the identical patterns with normal proliferative phase of endometrium. Only one case showing complex hyperplasia with atypia was focally positive for CD44s and CD44v6 in glandular epithelia. CD44s and CD44v6 were positive in all endometrial adenocarcinomas (13), except one CD44s-negative case. In summary, the expressions of CD44s and CD44v6 in endometriosis and adenomyosis recapitulated those of normal cyclic endometrium. The expression patterns in endometrial hyperplasia were similar to those in normal proliferative endometrium, whereas the endometrial adenocarcinoma showed abnormal expressions for CD44s and CD44v6. Thus it was considered that the ectopic endometrium in endometriosis and adenomyosis was not aberrant as in endometrial carcinoma on the aspects of immunohistochemical expressions of CD44s and CD44v6.

Expresion diferencial de los carbohidratos de mucina en endometrios humanos / Differential expression of mucin carbohydrates in human endometriuns

La aplicación de Lectinas para la identificación de oligosacáridos constituyentes de las Mucinas presentes en la superficie celular, es una herramienta técnica muy útil, debido a que, probablemente estas sustancias estén involucrados en procesos tales como invasión y metástasis. En este trabajo, nosotros estudiamos tejido endometrial normal com entidades benignas y malignas, para investigar la presencia de Galactosa beta 1-3 N Acetilgalactosamina (Galbeta 1-3GalNacalpha) y de Galactosa beta 1-3 N Acetilgalactosamina(Galbeta 1-3GalNa alpha y beta) empleando dos Lectinas: Agaricus bisporus (ABL) y Arachis hipogea (PNA) respectivamente. Lo controles fueron realizados com baños de galactosa para PNA y com mucina porcina de estómago de cerdo para ABL. El uso de estas dos Lectinas, permitió encontrar diferencias en los patrones de unión, ya que si bien ambas se unen a los mismos oligosacáridos, ABL realiza la unión en presencia de Acido Siálico mientras que PNA no. Se observaron significativas diferencias en los patrones de unión de ambas lectinas en tejidos, com entidades benignas, malignas y tejido normal. En este último, la marcación fue simpre continua com ambas Lectinas, mientras que fue irregular en el carcinoma.