Fabio Bagnoli

The interaction between aromatase, metalloproteinase 2,9 and cd44 in breast cancer

OBJECTIVE This study intends to verify the expression levels and correlation of aromatase, matrix metalloproteinase 2 (MMP-2), matrix metalloproteinase 9 (MMP-9) and CD44 in ductal carcinoma in situ (DCIS) and infiltrating ductal carcinoma (IDC) when both are found in the same breast. METHODS One hundred and ten cases were evaluated by tissue microarray (TMA) and immunohistochemically screened with anti-aromatase polyclonal antibodies, anti-MMP-2 monoclonal antibodies, anti-MMP-9 polyclonal antibodies and anti-CD44 monoclonal antibodies. RESULTS Aromatase was expressed in IDC and DCIS in 63 (57.3%) and 60 (67%) of the cases respectively; MMP-2 was similarly expressed in IDC and DCIS in 15 (13.60%) cases; MMP-9 was positively expressed in IDC and DCIS in 83 (75.50%) and 82 (74.50%) cases, respectively; CD44 was positively expressed in IDC and DCIS in 49 (44.50%) and 48 (42.60%) of the cases, respectively; all of them were highly correlated (p<0,001). The correlation analysis found positive, statistically significant correlation, in IDC between aromatase and MMP-2 (p<0.001) and between aromatase and MMP-9 (p=0.034). Positive correlation between aromatase and MMP-2 (p<0.001) and between MMP-9 and CD44 (p=0.030) were found in DCIS. CONCLUSION These results allow us to conclude that aromatase through local estrogen synthesis in breast tissue plays an important role in breast carcinogenesis, mainly influencing MMP-2 and MMP-9 which are important participants in tumor cell invasion and dependence of their connection to CD44 for action.

Oncoplastic Mammaplasty With Geometric Compensation-a Technique for Breast Conservation

To describe a technique of oncoplastic mammaplasty, referred to as geometric compensation, which is suitable for tumors close to the skin in areas not included in the classic preoperative drawings for mammaplasty.

Expressão da aromatase em carcinomas de mama ductais invasivo e in situ presentes na mesma 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%) and negatively in 13(29%) 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%) and negative in 26 (57.8%) 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%) showing no correlation with aromatase expression. Aromatase was positive in 36(80%) 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.

Oncoplastic Mammaplasty with Geometric Compensation – A Novel Technique for Breast Conservation

INTRODUCTION: Oncoplastic mammaplasties can prevent many mastectomies, but there are still limitations regarding their use for tumors close to the skin, in areas outside the classic mammaplasty drawings. This study describes a novel means of oncoplastic mammaplasty, named geometric compensation. MATERIAL AND METHODS: Between March 2007 and June 2012, thirteen patients were included in the study, after signing an informed consent statement. Six patients were from the Federal University of Goias, four from the private office, and three from the Santa Casa de Misericordia of Sao Paulo. They had malignant masses, needing skin resection in areas not usually resected in conventional mammaplasties. The preoperative markings followed the “Wise Pattern” technique, and the aberrant skin resection was geometrically compensated with another area of preserved skin. The tumors were excised with a macroscopic margin of 2 cm. The compromised quadrants were: superior in 5 (38.46%) cases, inferior in 4 (30.77%), and central in one (7.70%). Multiple quadrants were involved in 3 (23.08%) patients. The nipple-areola complex (NAC) was resected in 3 cases (23.08%), and it was kept attached to the superior pedicle in 5 (38.46%), to the inferior pedicle in 4 (30.77%) and to the lateral pedicle in 1 (7.70%), depending on the best available vascularization. Metal clips were emplaced to guide radiotherapy. The contralateral breast symmetry was corrected. RESULTS: The patients’ mean age was 53.46 (+ 11.24) years. The mean pathological tumor size was 44.00mm (+34,06mm). There were 5 (38.46%) locally advanced tumors (T3 or T4), the largest one measuring 140mm. Sentinel node biopsy was performed in 8 (61.54%) cases, all of them successfully. Three (23.07%) patients have lymph nodes with metastasis. Five (38.46%) patients were submitted to neoadjuvant and the remaining to adjuvant chemotherapy. The adjuvant radiotherapy was indicated in all cases. Eleven (84.61%) patients received hormone therapy (10 tamoxifen alone and one sequential aromatase inhibitor). The median preoperative ptosis was grade 2 (range: 1-3), and it was corrected in all cases. The volume and shape symmetry were considered adequate in all cases by surgeons and patients. Invasive ductal carcinoma was the commonest tumor, in 7 (53.84%) cases. One (7.70%) patient was smoker, five (38.46%) had high blood pressure, and one (7.70%) was diabetic. All tumors were excised with free margins. There was not any case of reoperation, hematoma, seroma or dehiscence. There was one (7.70%) small fat necrosis, and one (7.70%) enlarged scar. There was not any case of local or distant recurrence in a mean follow-up of 20.92 (+18.64) months. CONCLUSION: The technique allowed breast conservation in some difficult oncologic situations, which required large aberrant skin excisions, permitting free margins, ptosis correction, satisfactory symmetry and low rate of complications.

Correlation Between Mammographic Breast Density and Expression of 17ß Hydroxysteroid Desydrogenase Type 1

INTRODUCTION: High Mammographic density is a breast cancer risk factor and it is present over one third of cases of breast cancer. The 17s-hydroxysteroid dehydrogenase (17s-HSD) enzyme has possible local effects on breast tissue by stimulating growth of breast epithelial cells and is associated with mammographic density in women in pre and perimenopause. After a review in the literature, we see the importance of the enzyme 17sHSD1 in the etiology of breast cancer by both local and peripheral steroidogenesis, which may increase breast density. OBJECTIVE: For this reason we studied the correlation between mammographic breast density and immunohistochemical expression of 17s hydroxysteroid desydrogenase type 1 (17sHSD1). Casuistic and METHODS: 48 cases were evaluated with mammographic image classified by BI-RADS® system in four categories. The pattern of density 1 and 2 (measure breast tissue until 50%) was classified like low density and the pattern 3 and 4 (measure breast tissue upper 50%) like high density. Expression of the enzymes were evaluated by tissue microarray (TMA) screened with anti-17sHSD1 monoclonal rabbit antibodies (LS-C49955, Lifespan) diluted 1/200. Specimens were assigned scores on a scale from zero to three according to intensity of staining and number of stained cells. Scores zero and one were considered negative results and scores two and three indicated positive enzyme expression. RESULTS: In relation to breast density, in absolute numbers and percentages, we found the following results in category 1, 17 patients (35.42%), category 2, 17 patients (35.42%), category 3, 6 patients (12 50%) and category 4, 8 patients (16.67%). The 17sHSD1 expression was positive in 18 (37.5%). Statistical analysis showed no significant correlation between the expression of 17sHSD1 (p = 0.00048) with breast density (p = 0.666). No significant correlation between the expression of 17sHSD1 with breast density was found to be unchanged even when the analysis of the proposed subgroups (BMI greater than or less than 25, aged 50 years or less and high and low mammographic density). CONCLUSION: The results of our studies showed that there was no positive association between mammographic breast density and the expression of the enzyme 17sHSD1.

The Expression of the Aromatase Enzyme and the Tumoral Invasion Markers on the Primary Breast Cancer Tumor and on its Metastatic Lymph Nodes

INTRODUCTION: Every year, 22% of the new cancer cases are due to breast cancer. The prognostic factors for evaluation of these patients show a prediction of the overall survival or disease free survival. Among these factors, there are the ones well known as the axillary lymph nodes, size of the tumor, histological type and grade, nuclear grade and the presence of hormones receptors; there are other factors that are not as studied such as the aromatase enzyme and the cell invasion markers. The metalloproteinases (MMPs) are markers that facilitate the angiogenesis and promote the tumor progression. They are also related to the activation of the wall receptors CD44. As we know, the axillary lymph nodes metastasis is an important event on the metastatic process of the breast cancer and measuring these markers on these areas could give us important information to improve the treatment of these patients. OBJECTIVE: The objective of our study is evaluate the relation of the aromatase enzyme and the biologic markers of the neoplasic progression (MMP-2, MMP-9, CD44) with the invasive ductal carcinoma (IDC) and it’s axillary metastasis. MATERIAL AND METHODS: It’s a transversal study with 885 breast cancer treated patients. 41 cases satisfied the criterion of inclusion and exclusion. The histological and immunohistochemical analysis of the 41 blocks (including IDC and axillary lymph nodes) was done and processed by tissue microarray. Results: The expression of the biological markers on the IDC was as follow: 57, 3% had the aromatase enzyme; 13, 6% MMP-2; 75, 5% MMP-9 and 44, 5% CD44. and the axillary lymph nodes: 42, 5% had the aromatase enzyme; 55, 5% MMP-2; 90, 6% MMP-9 and 60, 2% CD44. There is a positive correlation statistically significant between aromatase and MMP-9 in the lymph node metastasis (p=0, 0001). There is a positive correlation between CD44 and MMP-2 (p=0,043) and with MMP-9 (p=0,023) in IDC. The expression of the MMP-2 in IDC showed a positive correlation with it s expression in the lymph node (p=0, 0001), what suggests that these enzyme may be involved on the cancer dissemination process. The expression of aromatase in IDC shows positive correlation with aromatase in the lymph node (p< 0, 0001). CONCLUSION: We concluded that there is a correlation between the expression of these biological markers in the tumor (IDC) and its axillary nodes affected by breast cancer metastasis.

Abstract 448: Expression of Glypican-3 (GPC3) in breast cancer tumors from Brazilian and Argentinean patients

The heparan sulfate proteoglycan Glypican-3 (GPC3) is involved in the signaling regulation of several growth factors. GPC3 is widely expressed in embryonic tissues, but disappears from most adult tissues, except for the mammary gland, among others. Several studies have involved GPC3 with cancer and some data indicate that this glypican is diminished in breast tumors. Previously, we have transfected the murine mammary tumor cell line LM3 (GPC3 negative) with the GPC3 complete cDNA sequence. We found that GPC3 reexpression is able to inhibit invasion and metastasis in vivo as well as to induce an in vitro EMT reversion, suggesting that GPC3 could act as a metastasis suppressor. The aim of the present work was to determine, in a prospective trial, whether GPC3 expression might be useful as a biomarker able to predict metastasis outcome in breast cancer patients. Our local access to a major source of breast human tissues from a representative Latin American population is an exceptional tool for the development of this new biomarker. A preliminary study was perform to analyze the expression of GPC3 at the mRNA level, by quantitative real time RT-PCR, in breast cancer samples (Brazilian n=15, Argentinean n=22) and peri-tumoral normal breast tissues (Brazilian n=16, Argentinean n=4). Even though the analyzed sample size was small, we found that both Brazilian and Argentinean tumor populations presented similar characteristics related to GPC3 expression. It was established a lower GPC3 level in tumor (n=37) than in peri-tumoral normal tissues (n=20) (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 448. doi:1538-7445.AM2012-448