Characterization of reactive stroma in prostate cancer: involvement of growth factors, metalloproteinase matrix, sexual hormones receptors and prostatic stem cells

ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.

Beneficial effects montelukast, cysteinyl-leukotriene receptor antagonist, on renal damage after unilateral ureteral obstruction in rats

Introductıon Ureteral obstruction is a common pathology and caused kidney fibrosis and dysfunction at late period. In this present, we investigated the antifibrotic and antiinflammatory effects of montelukast which is cysteinyl leukotriene receptor antagonist, on kidney damage after unilateral ureteral obstruction(UUO) in rats. Mateirıals and Methods 32 rats divided four groups. Group 1 was control, group 2 was sham, group 3 was rats with UUO and group 4 was rats with UUO which were given montelukast sodium (oral 10 mg/kg/day). After 14 days, rats were killed and their kidneys were taken and blood analysis was performed. Tubular necrosis, mononuclear cell infiltration and interstitial fibrosis scoring were determined histopathologically in a part of kidneys; nitric oxide(NO), malondialdehyde(MDA) and reduced glutathione(GSH) levels were determined in the other part of kidneys. Urea-creatinine levels were investigated at blood analysis. Statistical analyses were made by the Chi-square test and one-way analysis of variance (ANOVA). Results There was no difference significantly for urea-creatinine levels between groups. Pathologically, there was serious tubular necrosis and fibrosis in group 3 and there was significantly decreasing for tubular necrosis and fibrosis in group 4(p<0.005). Also, there was significantly increasing for NO and MDA levels; decreasing for GSH levels in group 3 compared the other groups(p<0.005). Conclusıon We can say that montelukast prevent kidney damage with antioxidant effect, independently of NO. .

Expressão gênica dos REα, REβ e PR em tumores mamários de cadelas por meio do q-PCR / REα, REβ and PR gene expression in mammary gland tumors of female dogs by q-PCR

A expressão de receptores de estrógeno (ER) e progesterona (PR) por meio da técnica de q-PCR foi avaliada em 26 cadelas portadoras de neoplasias mamárias e cinco cadelas sem afecções mamárias (grupo controle). Os resultados mostraram que os três grupos de animais estudados - com tumor maligno ou benigno e controle - expressaram receptores de estrógeno alfa, beta e progesterona. A quantificação relativa mostrou tendência para uma expressão maior de receptores no grupo controle e menor no grupo de animais com neoplasias malignas. Além disso, observou-se expressão maior de ERα em relação ao ERβ, e as neoplasias malignas de origem mista apresentaram maiores concentrações dos receptores PR, ERα e ERβ que as neoplasias de origem epitelial.

The estrogen and progesterone receptor (ER and PR) expression with the q-PCR technique was evaluated in 26 female dog carrying of mammary tumors and five female dogs without mammary disease (control group). The results showed that the three animal groups evaluated - malignant or benign tumor and control - expressed alpha and beta estrogen and progesterone receptors. The relative quantification showed a tendency for a higher expression of receptors from the control group and smaller in the malignat tumors animal group. Also, there was a major ERα expression regarding to ERβ and the malignat tumors from mixed origin presented higher concentrations of receptors PR, ERα and ERβ, when compared to tumors of epithelial origin.

Inmunohistochemical evaluation of estrogen receptors alpha and beta in normal inferior turbinate mucosa / Evaluación inmunohistoquímica de los receptores de estrógenos alfa y beta en la mucosa normal de concha nasal inferior

It has been postulated that the nasal mucosa, like other human tissues, is affected by a complex interactive network of neuropeptides, cytokines, allergic and inflammatory mediators and hormones such as estrogen, in which associations between symptoms (e.g. nasal stuffiness and coryza) and hormonal variations deriving from pregnancy, use of hormonal contraceptives and menstrual cycle phases are observed. The objective is evaluating the presence of specific estrogen receptors (types alpha and beta) in inferior turbinate mucosa in healthy subjects without nasal symptoms. Samples of nasal inferior turbinate were removed from patients undergoing aesthetic nasal surgery, and analyzed using hematoxylin-eosin staining, followed by immunohistochemical preparations on paraffin-embedded sections from the material sample, to detect estrogen receptors alpha and beta. Positive immunohistochemical reactions for both beta and alpha receptors were found in various regions of the inferior nasal turbinate. In conclusion both alpha and beta receptors were found, though the expression of beta was greater and more intense in the anterior portion of the inferior turbinate. No difference was found between male and female patients regarding the intensity of expression of receptors in the inferior turbinate.

Se ha postulado que la mucosa nasal, al igual que otros tejidos humanos, se ve afectada por una compleja red interactiva de neuropéptidos, citoquinas, mediadores alérgicos e inflamatorios, y hormonas como el estrógeno, en el que las asociaciones entre los síntomas (por ejemplo, congestión nasal y catarro) y hormonales las variaciones derivadas del embarazo, se observó el uso de anticonceptivos hormonales y las fases del ciclo menstrual. El objetivo es evaluar la presencia de receptores de estrógenos específicos (tipos de alfa y beta) en la mucosa de la concha nasal inferior en sujetos sanos sin síntomas nasales. Las muestras de la concha nasal inferior fueron retirados de los pacientes sometidos a cirugía nasal estética y analizados mediante hematoxilina-eosina, seguidos de cortes de preparados de inmunohistoquímica incluídos en parafina de la muestra de material, para detectar los receptores de estrógenos alfa y beta. Las reacciones de inmunohistoquímica fueron positiva para ambos receptores alfa y beta, éstas se encuentran en diversas regiones del cornete nasal inferior. En conclusión, tanto los receptores alfa y beta se encuentran, aunque la expresión de la beta fue mayor y más intensa en la porción anterior de la concha nasal inferior. No se encontraron diferencias entre pacientes hombres y mujeres en relación con la intensidad de la expresión de los receptores en el concha nasal inferior.

Differential binding with ERá and ERâ of the phytoestrogen-rich plant Pueraria mirifica

Variations in the estrogenic activity of the phytoestrogen-rich plant, Pueraria mirifica, were determined with yeast estrogen screen (YES) consisting of human estrogen receptors (hER) hERá and hERâ and human transcriptional intermediary factor 2 (hTIF2) or human steroid receptor coactivator 1 (hSRC1), respectively, together with the â-galactosidase expression cassette. Relative estrogenic potency was expressed by determining the â-galactosidase activity (EC50) of the tuber extracts in relation to 17â-estradiol. Twenty-four and 22 of the plant tuber ethanolic extracts interacted with hERá and hERâ, respectively, with a higher relative estrogenic potency with hERâ than with hERá. Antiestrogenic activity of the plant extracts was also determined by incubation of plant extracts with 17â-estradiol prior to YES assay. The plant extracts tested exhibited antiestrogenic activity. Both the estrogenic and the antiestrogenic activity of the tuber extracts were metabolically activated with the rat liver S9-fraction prior to the assay indicating the positive influence of liver enzymes. Correlation analysis between estrogenic potency and the five major isoflavonoid contents within the previously HPLC-analyzed tuberous samples namely puerarin, daidzin, genistin, daidzein, and genistein revealed a negative result.

Predictive factors of breast cancer evaluated by immunohistochemistry: [review] / Fatores preditivos do câncer de mama avaliados pela imuno-histoquímica: [revisão]

Hormone receptor and Her2 protein overexpression evaluated by immunohistochemistry (IHC) is widely validated as a predictive factor in breast cancer. The quality of the IHC reaction is influenced by tissue fixation and processing. Over- and underfixation deeply affect IHC results. Antigen retrieval may improve IHC but it does not recover tissue from autolysis or overfixation. The choice of primary antibody for IHC as to its sensitivity and specificity in relation to therapeutic response represents an important stage. Apart from mouse monoclonal antibodies, new rabbit monoclonal antibodies are commercially available, such as clones anti-ER SP1 and B644, anti-PR SP2 and B645 and anti-Her2 SP3 and 4B5. They represent an alternative to hormone receptor and Her2 evaluation by IHC. New polymeric non-biotinylated detection systems are also available and allow accurate and strong marking with no stromal and no non-specific cytoplasmic staining due to endogenous biotin. The most recommended cut off for estrogen and progesterone receptors (ER and PR) is more than 1 percent of positive cells with moderate or strong staining intensity (Allred's scoring system). New guidelines for Her2 evaluation by IHC show a cut off of more than 30 percent of positive cells with strong intensity (3+) that correlates better with gene amplification. The 2+ cases are now considered indeterminate and should be confirmed by fluorescence in situ hybridisation (FISH) or chromogenic in situ hybridisation CISH. A quality control of pre-analytical, analytical and post-analytical phases of IHC is recommended in order to optimize results.

A superexpressão de receptores hormonais e Her2 avaliada pela imuno-histoquímica (IHQ) é amplamente validada como fator preditivo em câncer de mama. A qualidade da reação imuno-histoquímica é influenciada pela fixação do tecido e seu processamento. A fixação insuficiente ou demasiada afeta profundamente os resultados da IHQ. A reativação antigênica pode melhorar os resultados da IHQ, porém não recupera tecidos com autólise ou com excessiva fixação. A escolha do anticorpo primário para a IHQ, considerando sua sensibilidade e sua especificidade de acordo com a resposta terapêutica, representa uma importante etapa. Além de anticorpos monoclonais de camundongo, novos anticorpos monoclonais de coelho são comercialmente disponíveis, tais como clones SP1 e B644 anti-RE, SP2 e B645 anti-RP, e SP3 e 4B5 anti-Her2. Eles representam uma alternativa para avaliação de receptores hormonais e Her2 através da IHQ. Novos sistemas de detecção poliméricos não-biotinilados também são disponíveis e permitem marcação exata e forte sem marcação estromal ou citoplasmática inespecífica devido à biotina endógena. O cut off mais recomendado para receptor de estrogênio (RE) e receptor de progesterona (RP) é acima de 1 por cento de células positivas com marcação moderada ou forte (sistema de escore de Allred). Novas recomendações para avaliação de Her2 através da IHQ apontam um cut off de mais de 30 por cento de células positivas com marcação forte (3+), que melhor se relaciona com amplificação gênica. Os casos 2+ são agora considerados indeterminados e devem ser confirmados por hibridação in situ por fluorescência (FISH) ou hibridização in situ colorimétrica (CISH). Um controle de qualidade de fases pré-analítica, analítica e pós-analítica da IHQ é recomendado para a otimização dos resultados.

Expression of the RERG Gene is Gender-Dependent in Hepatocellular Carcinoma and Regulated by Histone Deacetyltransferases

Ras-related, estrogen-regulated, and growth-inhibitory gene (RERG) is a novel gene that was first reported in breast cancer. However, the functions of RERG are largely unknown in other tumor types. In this study, RERG expression was analyzed in hepatocellular carcinomas of human patients using reverse transcriptase PCR analysis. In addition, the possible regulation of RERG expression by histone deacetyltransferases (HDACs) was studied in several cell lines. Interestingly, the expression of RERG gene was increased in hepatocellular carcinoma (HCC) of male patients (57.9%) but decreased in HCC of females (87.5%) comparison with paired peri-tumoral tissues. Moreover, RERG gene expression was increased in murine hepatoma Hepa1-6 cells, human breast tumor MDA-MB-231 cells, and mouse normal fibroblast NIH3T3 cells after treated by HDAC inhibitor, trichostatin A. Our results suggest that RERG may function in a gender-dependent manner in hepatic tumorigenesis and that the expression of this gene may be regulated by an HDAC-related signaling pathway.