Cyclooxygenase-2 and human epidermal growth factor receptor type 2 (HER-2) expression simultaneously in invasive and in situ breast ductal carcinoma / Expressão da ciclo-oxigenase-2 e receptor tipo 2 do fator de crescimento epidérmico humano (HER-2) simultaneamente em carcinoma ductal de mama in situ e invasivo

CONTEXT AND OBJECTIVE: Cyclooxygenase-2 (COX-2) and human epidermal growth factor receptor type 2 (HER-2) are associated with tumorigenesis. Studies have shown that HER-2 can regulate COX-2 expression. The aim of this study was to evaluate the correlation between COX-2 and HER-2 expression in normal breast epithelium and in ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) present in the same breast. DESIGN AND SETTING: Cross-sectional study at the Mastology Unit of the Department of Gynecology and Obstetrics, Santa Casa de Misericórdia de São Paulo Hospital. METHODS: COX-2 and HER-2 were detected using immunohistochemistry on 100 tissue fragments. HER-2 > +2 was subjected to fluorescence in situ hybridization (FISH). RESULTS: COX-2 expression was detected in 87 percent, 85 percent and 75 percent of IDC, DCIS and normal epithelium, respectively. HER-2 expression was detected in 34 percent of IDC and 34 percent of DCIS. COX-2 in DCIS correlated with HER-2 in IDC (P = 0.049) and DCIS (P = 0.049). COX-2 in normal epithelium correlated with HER-2 in IDC (P = 0.046) and DCIS (P = 0.046). COX-2 in IDC was not associated with HER-2 (P = 0.235). Comparison between COX-2 and HER-2 in DCIS showed that there was a statistically significant difference with regard to nuclear grades II and III and presence of comedonecrosis (P < 0.001). In IDC, there was significant expression with nuclear grades II and III and histological grade II (P < 0.001). CONCLUSIONS: Our findings provide evidence that HER-2 and COX-2 regulate each other.

CONTEXTO E OBJETIVO: Ciclo-oxigenase (COX-2) e receptor tipo 2 do fator de crescimento epidérmico humano (HER-2) estão associados com tumorigênese. Estudos mostraram que HER-2 pode regular a expressão de COX-2. O objetivo deste estudo foi avaliar a correlação entre expressão da COX-2 e HER-2 no epitélio normal de mama, no carcinoma ductal in situ (DCIS) e carcinoma ductal invasivo (IDC) presentes na mesma mama. TIPO DE ESTUDO E LOCAL: Estudo transversal na clínica de Mastologia do Departamento de Obstetrícia e Ginecologia do Hospital da Santa Casa de Misericórdia de São Paulo. MÉTODOS: A detecção da COX-2 e HER-2 foi realizada por imunoistoquímica em 100 fragmentos teciduais. HER-2 > +2 foi submetido a hibridização fluorescente in situ (FISH). RESULTADOS: Expressão de COX-2 foi detectada em 87 por cento, 85 por cento e 75 por cento dos IDC, DCIS e epitélio normal, respectivamente. Expressão de HER-2 foi detectada em 34 por cento dos IDC e 34 por cento de DCIS. COX-2 em DCIS correlacionou-se com HER-2 em IDC (P = 0,049) e DCIS (P = 0,049). COX-2 no epitélio normal correlacionou-se com HER-2 em IDC (P = 0,046) e DCIS (P = 0,046). COX-2 no IDC não foi associada com HER-2 (P = 0,235). Quando comparado COX-2 com HER-2 em DCIS houve diferença estatisticamente significante com relação ao grau nuclear II e III e presença de comedonecrose (P < 0,001) e no IDC, houve expressão significativa no grau nuclear II e III e histológico II (P < 0,001). CONCLUSÕES: Nossos achados mostram evidências que HER-2 e COX-2 se autorregulam.

Cox-2 e sua associação com fatores prognósticos e resposta à quimioterapia primária em pacientes com câncer de mama / Cox-2 and its association with prognostic factors and response to primary chemotherapy in patients with breast cancer

OBJETIVO: Avaliar em pacientes com câncer de mama a expressão imunoistoquímica da cox-2 antes da quimioterapia primária com 5-fluorouracil, epirrubicina e ciclofosfamida (FEC) e a associação desta com tamanho inicial do tumor, estado linfonodal, receptores hormonais, expressão da Her-2 e com a resposta clínica e anatomopatológica. MÉTODOS: Estudo retrospectivo com 41 mulheres portadoras do diagnóstico histopatológico de carcinoma ductal de mama. Foram submetidas à quimioterapia primária com esquema FEC (5-fluorouracil, epirrubicina e ciclofosfamida) na dosagem de 500mg/m2, 75mg/m2 e 500 mg/m2, respectivamente. Os critérios de inclusão foram intervalo etário entre 30 e 70 anos, estadiamento II a IIIA, após comprovação da ausência de metástase, tumor primário de mama, único e unilateral, tipo histológico ductal invasivo e ausência de cardiopatia e gestação. Para avaliação da expressão da proteína Her 2 neuutilizaram-se anticorpos monoclonais de coelho. Para visibilizar a expressão da proteína cox-2 utilizaram-se anticorpos policlonais obtidos do soro de cabras. A avaliação da resposta clínica ao tratamento foi realizada por exame físico mensurando-se o maior eixo tumoral por paquímetro. As medidas foram realizadas à admissão e após os ciclos de quimioterapia primária. Após três sessões quimioterápicas com intervalos de 21 dias realizou-se o procedimento cirúrgico. Adotaram-se os critérios do RECIST. Após a operação foi avaliada a resposta anatomopatológica local, sendo considerada completa quando da ausência de neoplasia invasiva e do componente in situ. Na avaliação imumoistoquímica para os receptores de estrogênio utilizaram-se estrogen receptor NCL-ER6F11 e para progesterona, progesterone receptor, NCL-PGR-312 considerando positiva quando da coloração em 10 por cento ou mais das células tumorais. RESULTADOS: A distribuição segundo estadiamento clínico UICC verificaram-se seis no estádio IIA (14,6 por cento), 22 no estádio IIB (53,6 por cento) e 13 estádio IIIA (31,8 por cento). A avaliação clínica inicial do maior eixo tumoral variou de 2,5 a 15 cm e mediana de 5 cm. Foram identificadas 14 pacientes (34,1 por cento) com estado linfonodal negativo e 27 positivo (65,9 por cento). Observou-se que 19 (46,3 por cento) apresentavam-se no menacme e 22 (53,6 por cento) na menopausa. CONCLUSÃO: Houve associação da expressão da cox-2 à fatores de pior prognóstico no câncer de mama como estado linfonodal positivo, receptores hormonais negativos e expressão da Her-2.

OBJECTIVE: To evaluate the immunohistochemical expression of cox-2 before primary chemotherapy with 5-fluorouracil, epirubicin and cyclophosphamide (FEC) and its association with initial tumor size, lymph node status, hormone receptors, expression of HER2 and the clinical and pathological response in patients with breast cancer. METHODS: We conducted a retrospective study with 41 women with histopathological diagnosis of ductal breast carcinoma. They underwent primary chemotherapy with FEC regimen (5-fluorouracil, epirubicin and cyclophosphamide) at 500mg/m2, 75mg/m2 and 500 mg/m2, respectively. Inclusion criteria were age range between 30 and 70 years, stage II to IIIA, absence of metastasis, primary tumor of the breast, single, unilateral, with ductal invasion at histology and absence of heart disease and pregnancy. To evaluate the expression of HER2/neu protein we used rabbit monoclonal antibodies. To visualize the expression of cox-2 protein we used polyclonal antibodies obtained from goats' serum. The evaluation of clinical response to treatment was performed during physical examination by measuring the major tumor axis with a pachymeter. Measurements were taken at admission and after primary chemotherapy cycles. After three chemotherapy sessions at intervals of 21 days the surgical procedure was carried out. We adopted the criteria of RECIST. After the operation we evaluated the local pathological response, which was considered complete when there was absence of invasive neoplasia and of the in situ component. In immunohistochemical assessing of estrogen receptors we used estrogen receptor NCL-ER6F11 and, for progesterone, progesterone receptor NCL-PGR-312, considering positive the staining of 10 percent or more tumor cells. RESULTS: The distribution according to UICC clinical stage classified six patients in stage IIA (14.6 percent), 22 in stage IIB (53.6 percent) and 13 stage IIIA (31.8 percent). The initial clinical evaluation of the major tumor axis ranged from 2.5 to 15 cm and a median of 5 cm. We identified 14 patients (34.1 percent) with negative lymph node status, and 27 positive (65.9 percent). It was observed that 19 (46.3 percent) were in premenopause and 22 (53.6 percent) in menopause. CONCLUSION: There was an association of the expression of Cox-2 to the factors associated with poor prognosis in breast cancer, such as positive lymph node status, negative hormone receptors and HER2 expression.

Clinical significance of matrix metalloproteinase 2 and 9 in breast cancer.

Background: Invasion and metastasis are the most strenuous problems in the management of breast cancer. These events require diverse proteolytic enzymes, among which MMP-2 and MMP-9 play a significant role in degradation of type IV collagen, the major component of the basement membrane. Therefore, the major objective of the study is to evaluate the clinical usefulness of MMP-2 and MMP-9 with respect to malignant tumor growth, invasion, and metastasis in breast cancer. Materials and Methods: Gelatin zymography was performed on 157 tissue extracts of malignant and adjacent normal breast tissues as well as negative and positive lymph nodes from 49 breast cancer patients. Statistical analysis was carried out using SPSS statistical software (version 10). Results: ProMMP-2 levels were significantly higher in adjacent normal tissues. Active MMP-2 and MMP-9 levels were higher in malignant breast tissues. Activation ratios of MMP-2 and MMP-9 were significantly higher in malignant breast tissues and in patients with lymph node metastasis. ProMMP-2, active MMP-2, and active MMP-9 could significantly discriminate between malignant and adjacent normal breast tissues. The MMP-2 activation ratio showed significant discriminatory efficacy between patients with and without lymph node metastasis and significant association with increased risk of lymph node metastasis in node-negative patients. Conclusion: The results indicate significant clinical utility of these proteolytic enzymes in malignant tumor growth, invasion, and metastasis in breast cancer.

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

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

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

Does thymidine phosphorylase correlate with angiogenesis in intraductal carcinoma of the breast?

To determine the association between thymidine phosphorylase [TP] and angiogenesis, and other conventional prognostic markers. We also evaluated interobserver and intraobserver reliability for TP expression in ductal carcinoma, to achieve a more consistent results. Our study included all cases diagnosed in Adnan Menderes University Medical Faculty Hospital, Aydin, Turkey as invasive ductal carcinoma or ductal carcinoma in situ [DCIS] with proven component of [>30%], between January 2003 and February 2005. The total number of the cases was 27 and their median age was 50 years. All sections were stained using monoclonal antibody-TP and examined at x40 magnification. Either nuclear or cytoplasmic staining was accepted as positive. The histoscore [H-score] was calculated for each specimen. The tumor stromal vascularity was assessed by monoclonal anti-CD34; and areas of intense vascularization were determined. Conventional immunohistochemical markers such as c-erb B2, Ki-67, estrogen and progesterone receptors and p53 were also applied to all slides. Three pathologists blindly examined each slide under 10 high-power fields [10 HPF] for 2 times in a 2 months period. There was no significant association between stromal vascularity and TP staining of cancer cells [p=0.1] and no correlation was determined between H-scores for TP staining in ductal carcinoma and DCIS components [p=0.5]. There was no significant correlation noted between stromal and periductal vascularity with the anti-CD34 antibody was used. No significant correlation was identified between the TP H-score and stromal or periductal vascularity