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.

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.

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.

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.

Frequencies of interleukin-6, GST and progesterone receptor gene polymorphisms in postmenopausal women with low bone mineral density / Frequência do polimorfismo da interleucina-6, GST, e dos receptores de progesterona em mulheres na pós-menopausa com baixa densidade mineral óssea

CONTEXT AND OBJECTIVE: Osteoporosis is a skeletal disorder characterized by low bone mineral density (BMD). Studies have shown that some of the genetic components relating to lower BMD may be detected by polymorphisms. Our aim was to evaluate the frequencies of interleukin-6, GST and progesterone receptor gene polymorphisms in postmenopausal women with low BMD. DESIGN AND SETTING: Cross-sectional study, conducted in a public university in São Paulo, Brazil. METHODS : We evaluated interleukin-6 (IL-6), progesterone receptor gene (PROGINS) and glutathione S-transferase (GST) polymorphisms in 110 postmenopausal women with no previous use of hormone therapy. Tests were performed using DNA-PCR, from oral scrapings. We used Student's t-test and a logistic regression model for statistical analysis. RESULTS : Regarding IL-6 polymorphism, 58.2% of the patients were homozygotes (GG) and 41.8% had allele C (heterozygote or mutant homozygote + GC or CC). PROGINS genotype polymorphism was absent in 79% (wild homozygote or P1/P1) and present in 20.9% (heterozygote or P1/P2). Regarding GSTM1 polymorphism, the allele (1/1) was present in 72.7% of the patients and was absent in 27.3%. We found that IL-6 polymorphism had statistically significant correlations with the L2-L4 T-score (P = 0.032) and with BMD (P = 0.005). Women with IL-6 polymorphism were 2.3 times more likely to have a L2-L4 T-score of less than -1, compared with those not presenting this polymorphism. CONCLUSION: IL-6 gene polymorphism was correlated with low BMD, whereas the PROGINS and GSTM1 polymorphisms did not show any correlation. .

CONTEXTO E OBJETIVO: A osteoporose é uma desordem esquelética caracterizada por baixa densidade mineral óssea. Estudos têm demonstrado que alguns componentes genéticos relacionados com a menor densidade mineral óssea podem ser detectados por polimorfismos. Nosso objetivo foi avaliar a presença do polimorfismo de genes em mulheres pós-menopáusicas com baixa densidade mineral óssea. TIPO DE ESTUDO E LOCAL: Estudo transversal, conduzido em universidade pública em São Paulo, Brasil. MÉTODOS: Avaliamos os polimorfismos relacionados à interleucina-6 (IL-6), o gene receptor de progesterona (PROGINS) e glutationa S-transferase (GST) em 110 mulheres na pós-menopausa sem terapia hormonal prévia. Os testes foram realizados com DNA-PCR a partir de raspados orais. Foram utilizados teste t de Student e modelo de regressão logística para análise estatística. RESULTADOS: Em relação ao polimorfismo IL-6, 58,2% dos pacientes eram homozigotos (GG) e 41,8% tinham o alelo C (heterozigoto ou homozigoto mutante + GC ou CC). Nos genótipos do polimorfismo PROGINS, 79% estavam ausentes (homozigoto selvagem ou P1/P1) e 20,9% presentes (heterozigoto ou P1/P2). No polimorfismo do GSTM1, o alelo (1/1) estava presente em 72,7% dos pacientes e ausente em 27,3%. Encontramos significância estatística entre o polimorfismo genético da IL-6 com o T-score de L2-L4 (P = 0,032) e a densidade mineral óssea (P = 0,005). As mulheres com polimorfismo da IL-6 tiveram 2,3 vezes mais chance de ter menos de -1 na L2-L4 T-score, quando comparadas às não portadoras. CONCLUSÃO: O polimorfismo do gene da IL-6 está correlacionado com baixa densidade mineral óssea, enquanto os polimorfismos GSTM1 e PROGINS não mostraram correlação. .

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.

Prevalence of ERα-397 PvuII C/T, ERα-351 XbaI A/G and PGR PROGINS polymorphisms in Brazilian breast cancer-unaffected women

Polymorphisms of hormone receptor genes have been linked to modifications in reproductive factors and to an increased risk of breast cancer (BC). In the present study, we have determined the allelic and genotypic frequencies of the ERα-397 PvuII C/T, ERα-351 XbaI A/G and PGR PROGINS polymorphisms and investigated their relationship with mammographic density, body mass index (BMI) and other risk factors for BC. A consecutive and unselected sample of 750 Brazilian BC-unaffected women enrolled in a mammography screening program was recruited. The distribution of PGR PROGINS genotypic frequencies was 72.5, 25.5 and 2.0% for A1A1, A1A2 and A2A2, respectively, which was equivalent to that encountered in other studies with healthy women. The distribution of ERα genotypes was: ERα-397 PvuII C/T: 32.3% TT, 47.5% TC, and 20.2% CC; ERα-351 XbaI A/G: 46.3% AA, 41.7% AG and 12.0% GG. ERα haplotypes were 53.5% PX, 14.3% Px, 0.3% pX, and 32.0% px. These were significantly different from most previously published reports worldwide (P < 0.05). Overall, the PGR PROGINS genotypes A2A2 and A1A2 were associated with fatty and moderately fatty breast tissue. The same genotypes were also associated with a high BMI in postmenopausal women. In addition, the ERα-351 XbaI GG genotype was associated with menarche ≥12 years (P = 0.02). ERα and PGR polymorphisms have a phenotypic effect and may play an important role in BC risk determination. Finally, if confirmed in BC patients, these associations could have important implications for mammographic screening and strategies and may be helpful to identify women at higher risk for the disease.

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.

Pólipos endometriais: aspectos clínicos, epidemiológicos e pesquisa de polimorfismos / Endometrial polyps: clinical and epidemiological aspects and analysis of polymorphisms

OBJETIVO: avaliar as variáveis clínicas e epidemiológicas de risco para câncer de endométrio em mulheres com pólipos endometriais na pós-menopausa, bem como a presença do polimorfismo do receptor da progesterona (PROGINS). MÉTODOS: estudo caso-controle desenhado com 160 mulheres na pós-menopausa com pólipos endometriais, comparado a Grupo Controle de 400 mulheres na pós-menopausa. A genotipagem do polimorfismo PROGINS foi determinada pala reação em cadeia da polimerase (PCR). Aspectos clínicos e epidemiológicos foram comparados entre as mulheres com pólipos endometriais benignos e 118 dos controles normais. Estas variáveis foram também comparadas entre mulheres com pólipos benignos e pólipos malignos. RESULTADOS: a comparação entre o grupo de pólipos benignos e o Grupo Controle mostrou diferença significativa (p<0,05) para as varáveis: idade, raça não-branca, anos da menopausa, paridade, hipertensão arterial, uso de tamoxifeno e antecedente de câncer de mama, todas mais prevalentes no grupo de pólipos endometriais. Após o ajuste para a idade, permaneceram com diferença significativa a paridade (OR=1,1), hipertensão arterial (OR=2,2) e o antecedente de câncer de mama (OR=14,4). Houve seis casos de pólipos malignos (3,7 por cento). A frequência de sangramento para pólipos benignos e malignos foi de 23,4 e 100 por cento, respectivamente, sendo o pólipo grande encontrado em 54,5 por cento dos casos benignos e em 100 por cento dos malignos. A frequência de hipertensão arterial foi de 54,5 por cento para pólipos benignos e 83,3 por cento para pólipos malignos. As frequências do polimorfismo PROGINS T1/T1, T1/T2 e T2/T2 foram 79,9 por cento, 19,5 por cento e 0,6 por cento respectivamente para pólipos benignos e 78,8 por cento, 20,8 por cento e 0,5 por cento para o Grupo Controle. CONCLUSÕES: os pólipos endometriais se mostraram mais frequentes em mulheres de idade avançada, hipertensas e com antecedente de câncer de mama. A presença do polimorfismo PROGINS não mostrou associação significativa com pólipos endometriais. A incidência de pólipos malignos foi baixa, estando fortemente associada à presença de sangramento, tamanho grande do pólipo e hipertensão arterial.

PURPOSE: to evaluate the clinical and epidemiological risk factors for endometrial cancer in postmenopausal women with endometrial polyps, as well as the genetic polymorphism of the progesterone receptor (PROGINS). METHODS: a case-control study was designed with 160 postmenopausal women with endometrial polyps, compared to a normal Control Group of 400 postmenopausal women. The genotyping of PROGINS polymorphism was determined by the polymerase chain reaction. Clinical and epidemiological data were compared between benign endometrial polyps and 118 of the control subjects. Variables were also compared with regard to benign and malignant endometrial polyps. RESULTS: comparison of the epidemiological variables between groups showed a significant difference for age, ethnicity, time since menopause, parity, tamoxifen use, hypertension and breast cancer, all of them more prevalent in the polyp group. After adjustment for age, statistical significance remained only for parity (OR=1.1), hypertension (OR=2.2) and breast cancer (OR=14.4). There were six cases of malignant polyps (3.7 percent). The frequency of bleeding was 23.4 percent for benign polyps and 100 percent for malignant polyps, with large polyps being detected in 54.6 percent of the benign cases and in 100 of the malignnat ones. The frequency of arterial hypertension was 54.5 percent for benign polyps and 83.3 percent for the malignant ones. The frequency of PROGINS T1/T1, T1/T2 and T2/T2 polymorphism was 79.9 percent, 19.5 percent and 0.6 percent, respectively, for the polyp group, and 78.8 percent, 20.8 percent and 0.5 percent for the Control Group. CONCLUSIONS: elderly age, hypertension, and breast cancer were significantly associated with endometrial polyps. The presence of PROGINS polymorphism was not significantly associated with endometrial polyps. The incidence of malignant polyps was low and strongly associated with bleeding, large-sized polyp and arterial hypertension.

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.

Polimorfismo do gene dos receptores de progesterona e o aborto espontâneo de repetição / Progesterone receptor gene polymorphism and recurrent spontaneous abortion

OBJETIVO: investigar se polimorfismos dos genes que codificam o receptor de progesterona (PROGINS) estão relacionados à ocorrência de aborto espontâneo de repetição (AER). MÉTODOS: em estudo caso-controle, foram selecionados 85 pacientes com antecedente de pelo menos três abortos precoces sem etiologia definida (Grupo Caso) e 157 mulheres com história de pelo menos duas gestações de termo sem intercorrências e sem passado de abortamento (Grupo Controle). Realizada coleta de 10 mL de sangue por punção venosa periférica e extração de DNA pela técnica DTAB/CTAB. As genotipagens foram feitas por reação em cadeia de polimerase (PCR), nas condições de ciclagem específica para o polimorfismo em estudo, seguida de amplificação em gel de agarose a 2 por cento. A visualização das bandas foi feita sob luz ultravioleta e os géis foram fotografados. As diferenças genotípicas e alélicas entre os dois grupos para o polimorfismo PROGINS foram calculadas pelo teste de χ2, adotando-se como nível de significância valores de p<0,05. Calculou-se ainda o Odds Ratio (OR, razão de chances), com intervalos de confiança de 95 por cento (IC95 por cento). RESULTADOS: As frequências genotípicas encontradas para o polimorfismo PROGINS foram de 72,3 por cento T1/T1 e 27,7 por cento T1/T2 no grupo com AER e 76,4 por cento T1/T1, 22,3 por cento T1/T2 e 1,3 por cento T2/T2 no Grupo Controle. Não houve diferenças entre os grupos, analisando-se as frequências genotípicas e alélicas: respectivamente p=0,4 (OR: 0,8) e p=0,6 (OR: 0,9). CONCLUSÕES: os dados do presente estudo sugerem que o polimorfismo PROGINS do gene dos receptores de progesterona não está relacionado ao AER.

PURPOSE: to assess a possible association between polymorphism of the progesterone receptor gene (PROGINS) and recurrent spontaneous abortion (RSA). METHODS: in this case-control study, 85 women with at least three previous spontaneous abortions without an identifiable cause (RSA Group) and 157 women with at least two previous term pregnancies without pathologies and no previous miscarriage (Control Group) were selected. An amount of 10 mL of peripheral blood was collected by venipuncture and genomic DNA was extracted by the DTAB/CTAB method, followed by the polymerase chain reaction (PCR) under specific conditions for this polymorphism and by amplification by 2 percent agarose gel electrophoresis. The bands were visualized with an ultraviolet light transilluminator and the gels were photographed. Differences in the PROGINS genotype and allele frequencies between groups were analyzed by the χ2 test, with the level of significance set at p<0.05. The Odds Ratio (OR) was also used, with 95 percent confidence intervals 95 percentCI. RESULTS: PROGINS genotypic frequencies were 72.3 percent T1T1 and 27.7 percent T1T2 for the RSA group and 764 percent T1T1, 22.3 percent T1T2 and 1.3 percent T2T2 for the control group. There were no differecnes between groups when the genotype and allele frequencies were analyzed: respectively p=0.48 (OR: 0.8) and p=0.65 (OR: 0.9). CONCLUSIONS: our results suggest that PROGINS polymorphism is not associated with RSA.

The progins progesterone receptor gene polymorphism is not related to endometriosis-associated infertility or to idiopathic infertility

OBJECTIVE: This study aimed to determine the frequency of the PROGINS polymorphism in women with endometriosis-associated infertility, in infertile women without endometriosis and in controls. INTRODUCTION: The human progesterone receptor gene has two isoforms that modulate the biological action of progesterone: isoform A, which is capable of inhibiting the activation of the estrogen receptors, and isoform B, which has the capacity to activate the estrogen receptors. Several polymorphisms have been described for this gene, among which one stands out: a polymorphism named PROGINS, which has been speculated to be related to the genesis of endometriosis by several studies with conflicting results. METHODS: This was a prospective study that included 148 patients with endometriosis-associated infertility, 50 idiopathic infertile patients and 179 fertile women as controls. The PROGINS polymorphism was studied by PCR. RESULTS: Genotypes P1P1, P1P2 and P2P2 (P2 representing the PROGINS polymorphism) of the progesterone receptor gene presented frequencies of 93.9 percent, 5.4 percent and 0.7 percent, respectively, in the women with endometriosis-associated infertility (p=0.2101, OR=0.51, 95 percent CI=0.24-1.09); 94.4 percent, 4.2 percent and 1.4 percent, respectively, in the patients with minimal/mild endometriosis (p=0.2725, OR=0.53, 95 percent CI=0.20-1.43); 93.5 percent, 6.5 percent and 0 percent, respectively, among the patients with moderate/severe endometriosis (p=0.3679, OR=0.49, 95 percent CI=0.18-1.31); 86.0 percent, 14.0 percent and 0 percent, respectively, in idiopathic infertile women (p=0.8146, OR=1.10, 95 percent CI=0.46-2.63); and 88.3 percent, 10.6 percent and 1.1 percent, respectively, in the control group. CONCLUSION: The data suggest that PROGINS is not related either to endometriosis-associated infertility or to idiopathic infertility in the population studied.

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.

Polymorphism in CYP17, GSTM1 and the progesterone receptor genes and its relationship with mammographic density

Radiologic breast density is one of the predictive factors for breast cancer and the extent of the density is directly related to postmenopause. However, some patients have dense breasts even during postmenopause. This condition may be explained by the genes that codify for the proteins involved in the biosynthesis, as well as the activity and metabolism of steroid hormones. They are polymorphic, which could explain the variations of individual hormones and, consequently, breast density. The constant need to find markers that may assist in the primary prevention of breast cancer as well as in selecting high risk patients motived this study. We determined the influence of genetic polymorphism of CYP17 (cytochrome P450c17, the gene involved in steroid hormone biosynthesis), GSTM1 (glutathione S-transferase M1, an enzyme involved in estrogen metabolism) and PROGINS (progesterone receptor), for association with high breast density. One hundred and twenty-three postmenopausal patients who were not on hormone therapy and had no clinical or mammographic breast alterations were included in the present study. The results of this study reveal that there was no association between dense breasts and CYP17 or GSTM1. There was a trend, which was not statistically significant (P = 0.084), towards the association between PROGINS polymorphism and dense breasts. However, multivariate logistic regression showed that wild-type PROGINS and mutated CYP17, taken together, resulted in a 4.87 times higher chance of having dense breasts (P = 0.030). In conclusion, in the present study, we were able to identify an association among polymorphisms, involved in estradiol biosyntheses as well as progesterone response, and radiological mammary density.

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.

Selective expression of progesterone receptor in malignant melanoma was inversely correlated with PCNA / 华中科技大学学报（医学）（英德文版）

To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.

Hormone receptor expression and Her/2neu amplification in breast carcinoma in a cohort of Sri Lankans.

OBJECTIVE: 1) To determine the profile of estrogen and, progesterone receptors (ER, PR) expression and Her/2neu amplification in carcinoma of breast of Sri Lankan women. 2) To determine their inter-relationships, and associations with age at diagnosis and histological grade. DESIGN: Retrospective analysis of data. SETTING: Department of Pathology, Faculty of Medicine, University of Peradeniya. MATERIALS: 124 samples of invasive ductal carcinoma of breast, stained for steroid receptors and Her/2neu amplification with immunohistochemistry. MEASUREMENTS: 1) Semiquantative scores of steroid receptors and Her/2neu amplification. 2) Correlations of ER, PR, Her/2neu amplification, age at diagnosis and histological grade. RESULTS: The prevalence of ER, PR and Her/2neu amplification were 53.2%, 50% and 14.6% respectively. The expression of ER and PR were significantly correlated (p < 0.001). and had a significant negative correlation with Her/2neu amplification (p0.003 each). Lower grade of the tumour was significantly related to the expression of ER (p0.000) and PR (p0.000) but not to Her/2neu amplification (p0.331). Age at diagnosis was significantly correlated to the expression of ER (p0.004), but not to PR (p0.365) or Her/2neu amplification (p0.456). CONCLUSION: Prevalence or ER, PR and Her/2neu amplification in carcinoma of breast among Sri Lankans is similar to that described internationally. The correlations of ER, PR, Her/2neu amplification, to each other, age at diagnosis and grade of tumour is as reported in other countries.