Are both distinct epithelial and stromal cells molecular analysis from phyllodes tumors versus fibroadenoma components affected in breast fibroepithelial progression?

PURPOSE: To determine molecular events involved in the tumorigenesis of phyllodes tumors (PT) and the role of each stromal (SC) and epithelial (EC) cell. METHODS: Frozen breast samples enriched with epithelial and stromal cells from three fibroadenomas and 14 PT were retrieved and laser microdissected. Sanger and polymerase chain reaction-based sequencing of exon 2 MED12 and TERT promoter hotspot mutations were performed; 44K microarray platform was used to analyze gene expression. RESULTS: All three fibroadenomas (FAs) presented mutations in MED12, but not in TERT, whose mutation was observed in five of the 14 PTs. EC and SC of each affected tumor displayed identical alterations. Of the total differentially expressed genes (DEG) (EC = 1,543 and SC = 850), 984 were EC-eDEGs and 291 were SC-eDEGs. We found a high similarity of diseases and functions enriched by both cell types, but dissimilarity in the number of enriched canonical pathways. Three signaling canonical pathways overlapping with EC and SC were predicted to be activated in one cell type and inactivated in the other, while no overlap in eDEGs was assigned to them. We also identified 13 EC-eDEGs and five SC-eDEGs enriched networks, in which the SC-eDEGs were able to segregate FA from PT samples. CONCLUSIONS: Identical TERT mutations from both SC and ES origins might affect the PTs tumorigenesis. Gene expression differences suggest coordinated molecular processes between these components with determinant differences acquired by SC, able to fully distinguish PTs from FAs lesions.

Immunoexpression of HER2 pathway related markers in HER2 invasive breast carcinomas treated with trastuzumab.

PURPOSE: We evaluated the immunoexpression of potential markers involved in the HER2 pathway in invasive breast carcinoma with HER2 amplification treated with trastuzumab. METHODS: Samples of ninety patients diagnosed and treated at two public Brazilian hospitals with overexpressed invasive carcinoma between 2009 and 2018 were included. Several markers (Bcl-2, CDK4, cyclin D1, EGFR, IGF1, IGF-1R, MDM2, MUC4, p16, p21, p27, p53, PTEN, RA, TNFα, and VEGF) were immune analyzed in the tumor by immunohistochemistry and then correlated with clinicopathological variables. RESULTS: Tumor sample expression results determined potential markers of good prognosis with statistically significant values: cyclin D1 with a nuclear grade, and recurrence; IGF-1 with tumor size, and death; p16 with a response after treatment; PTEN with a response after treatment, and death. Markers of poor prognosis: p53 with histological, and nuclear grade; IGF-1R with a compromised lymph node. The treatment resistance rate after trastuzumab was 40%; the overall survival was 4.13 years (95% CI 5.1-12.5) and the disease-free survival was 3.6 years (95% CI 5.1-13.1). CONCLUSIONS: The tumor samples profile demonstrated that cyclin D1, IGF-1, p16, and PTEN presented the potential for a good prognosis and p53 and IGF-1R for worse.

Ultrasound Measurement of the Distance between the Breast Tumor and the Skin: A Cut-Off Value for Safe Skin Preservation - Diagnostic Accuracy Study.

Background: In superficial tumors of the breast, it is necessary to plan the thickness of surgical skin flaps, and whether skin can be preserved for esthetics results. This study aimed to find an ultrasound-measured cut-off distance between tumor and skin (TSD) that allows patients to have the skin over the tumor spared. Methods: This is a diagnostic accuracy study comparing preoperative ultrasound TSD with pathological TSD and the thickness of the skin flaps. We recruited all consecutive women diagnosed with breast cancer between January 2017 and December 2019 whose surgical planning allowed to have the tumor and overlying skin to be removed in bloc (reconstruction procedures, situations where skin removal would not lead to esthetic problems, and superficially located tumors). Measurements were made: preoperatively (by ultrasound), during surgery (using a metal caliper to obtain the thickness of surgical skin flap), and after surgery (pathological). A pathological tumor-skin distance greater than surgical skin flap thickness would indicate preservation of skin above the tumor. Results: We evaluated 95 consecutive patients with 102 lesions. The average surgical flap thickness was 5.5 mm (3-10 mm). The ultrasound-measured cut-off TSD of 2.1 mm obtained 96.0% accuracy in predicting free anterior margin, considering a 5-mm-thick surgical flap. Conclusion: In breast superficial tumors, a cut-off distance of 2.1 mm or more measured preoperatively by ultrasound allows safe preservation of the skin above the tumor. Future studies need to follow up for longer the women submitted to skin preservation surgeries, especially those not undergoing radiotherapy.

Are both distinct epithelial and stromal cells molecular analysis from phyllodes tumors versus fibroadenoma components affected in breast fibroepithelial progression?

Purpose: To determine molecular events involved in the tumorigenesis of phyllodes tumors (PT) and the role of each stromal (SC) and epithelial (EC) cell. Methods: Frozen breast samples enriched with epithelial and stromal cells from three fibroadenomas and 14 PT were retrieved and laser microdissected. Sanger and polymerase chain reaction-based sequencing of exon 2 MED12 and TERT promoter hotspot mutations were performed; 44K microarray platform was used to analyze gene expression. Results: All three fibroadenomas (FAs) presented mutations in MED12, but not in TERT, whose mutation was observed in five of the 14 PTs. EC and SC of each affected tumor displayed identical alterations. Of the total differentially expressed genes (DEG) (EC = 1,543 and SC = 850), 984 were EC-eDEGs and 291 were SC-eDEGs. We found a high similarity of diseases and functions enriched by both cell types, but dissimilarity in the number of enriched canonical pathways. Three signaling canonical pathways overlapping with EC and SC were predicted to be activated in one cell type and inactivated in the other, while no overlap in eDEGs was assigned to them. We also identified 13 EC-eDEGs and five SC-eDEGs enriched networks, in which the SC-eDEGs were able to segregate FA from PT samples. Conclusions: Identical TERT mutations from both SC and ES origins might affect the PTs tumorigenesis. Gene expression differences suggest coordinated molecular processes between these components with determinant differences acquired by SC, able to fully distinguish PTs from FAs lesions.

Entendendo o câncer de mama: educação em saúde / Understanding breast cancer: health education

Objetivo: Demonstrar os benefícios da educação em saúde sobre o câncer de mama para a população. Métodos: Trata-se de um relato de experiência sobre educação em saúde, utilizando boletins informativos e banners elaborados pelos discentes, monitorado pelos alunos de pós-graduação em patologia e supervisionados pelos colaboradores e coordenadores do projeto, divulgados presencialmente para a comunidade. Resultados: Essa experiência trouxe como resultado maior aproveitamento das atividades didáticas e dinâmicas do ambulatório, com uma metodologia ativa, assim como o empoderamento dos usuários para a autonomia, capacidade de identificar determinantes para sua saúde e cuidar de si e dos outros a sua volta. Conclusão: Conclui-se então que esta atividade de educação foi enriquecedora tanto para os discentes que a organizaram quanto para clientes do ambulatório, expondo uma relação entre a prevenção e a promoção da saúde dos usuários e com um projeto de extensão que visa compartilhar o conhecimento adquirido na academia, transcendendo seus muros para todos. (AU)

Objective: To demonstrate the benefits of health education on breast cancer for the population. Methods: Descriptive-exploratory study with a qualitative approach, using newsletters and banners prepared by students, monitored by graduate students in pathology and supervised by project collaborators and coordinators and disseminated to the community. Results: This experience resulted in greater use of the didactic and dynamic activities of the clinic, with an active methodology, as well as the empowerment of users for autonomy, the ability to identify determinants for their health and to take care of themselves and others around them. Conclusion: We conclude then that the campaign was enriching both for the students who organized it and for clients of the outpatient clinic, exposing a relationship between prevention and health promotion of users and with an extension project that aims to share the knowledge acquired in the academy, transcending their walls for everyone. (AU)

Objetivo: Demostrar los beneficios de la educación para la salud sobre el cáncer para la población. Métodos: Estudio descriptivo-exploratorio con un enfoque cualitativo, utilizando boletines y pancartas preparados por estudiantes, monitoreados por estudiantes graduados en patología y supervisados por colaboradores y coordinadores del proyecto y entregado a la comunidad. Resultados: Esta experiencia resultó en un mayor uso de las actividades didácticas y dinámicas de la clínica, con una metodología activa, así como el empoderamiento de los usuarios para la autonomía, la capacidad de identificar determinantes para su salud y para cuidarse a sí mismos y a los demás a su alrededor. Conclusión: Concluimos entonces que la campaña fue enriquecedora tanto para los estudiantes que la organizaron como para los clientes de la clínica ambulatoria, exponiendo una relación entre prevención y promoción de la salud de los usuarios y con un proyecto de extensión que tiene como objetivo compartir el conocimiento adquirido en la academia, trascendiendo sus muros para todos. (AU)

The Highly Expressed FAM83F Protein in Papillary Thyroid Cancer Exerts a Pro-Oncogenic Role in Thyroid Follicular Cells.

Thyroid cancer is the most common endocrine cancer with predominant prevalence of papillary thyroid cancer (PTC) histotype. MAPK signaling genetic alterations are frequent in PTC, affecting more than 80% of cases. These alterations constitutively activate MAPK signaling cross-regulating different pro-oncogenic pathways. However, additional molecular alterations associated with thyroid cancer are not completely understood. In this extent, the new family of proteins named FAM83 (FAMily with sequence similarity 83) was recently identified as mediator of oncogenic signaling in different types of cancer. Here we report FAM83F as a novel highly expressed protein in PTC. We evaluated FAM83F levels in 106 PTC specimens, 34 goiter, and 41 adjacent non-tumoral human thyroid, and observed FAM83F cytoplasmic overexpression in 71% of PTC (76 of 106) while goiter tissues showed nuclear positivity and normal thyroid showed no staining by immunohistochemistry. Moreover, TSH-induced goiter and BRAF T1799A -induced PTC animal models also showed FAM83F activation. In vitro, we generated a stable thyroid cell line PCCL3 with FAM83F overexpression and observed that FAM83F deregulates thyroid follicular cell biology leading to loss of thyroid differentiation genes such as Sodium-Iodide Symporter (NIS), reactivation of stem cell markers such as LIN28B and SOX2, induction of cell migration and resistance to doxorubicin-induced apoptosis. Moreover, FAM83F activates MAPK signaling through interaction with BRAF and RAF while impairs TGFß antiproliferative signaling transduction. In this study, we showed FAM83F as a new pro-oncogenic protein overexpressed in thyroid cancer that modulates thyroid follicular cell biology and differentiation through cross-regulation of MAPK and TGFß signaling.

MYC is expressed in the stromal and epithelial cells of primary breast carcinoma and paired nodal metastases.

The MYC oncogene is directly involved in the proliferation, metabolism, progression and distant metastasis of breast cancer. Since metastatic spread to the lymph nodes is often the first indication of propensity for metastatic dissemination, the MYC status in nodal disease may represent a decision-making variable. However, the analysis of MYC expression in stromal cells, namely cancer-associated fibroblasts (CAFs), which are known to play a critical role in cancer progression, remains poorly reported. The aim of this study was to determine the expression of MYC and other markers, including estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), p53, Ki67, epidermal growth factor receptor (EGFR), phosphorylated AKT (p-AKT) and phospho-mammalian target of rapamycin (p-mTOR) by immunohistochemistry in representative samples from 80 patients with ductal infiltrative breast cancer and 43 paired compromised axillary lymph nodes allocated in tissue microarrays (TMAs). The epithelial and stromal components of primary tumors and respective lymph node metastases were separately analyzed. MYC expression (cytoplasmic and nuclear) was a frequent event in the epithelial and stromal components of the primary tumors. The epithelial cells in the nodal metastases exhibited a trend for decreased MYC expression compared to that in the primary tumors (P=0.08) but retained the original status of the primary tumors for all other markers. The stromal cells were uniformly negative for ER, PR, HER2, p53, Ki67 and EGFR. Comparison of the stromas of primary tumors and respective lymph node metastases revealed a reduced frequency of nuclear MYC in 15% of the cases (P=0.003), whereas p-mTOR followed a similar trend (P=0.09). Analyses of the possible correlations among markers revealed that epithelial nuclear MYC was associated with p53 (P=0.048). This is an original study demonstrating a significant proportion of MYC expression (nuclear or cytoplasmic), as well p-mTOR and p-AKT expression, in the epithelial and stromal components of either the primary tumor or the nodal metastases. CAFs expressing MYC may establish an angiogenic microenvironment supporting cancer survival and facilitating colonization at the nodal metastatic site.

Integration of collagen matrices into the urethra when implanted as onlay graft.

OBJECTIVE: To assess the integration of decellularized heterologous collagen matrices into the urethra, when implanted with no cells or when seeded with autologous smooth muscle cells. MATERIALS AND METHODS: Eighteen New Zealand rabbits were randomly assigned to two groups: Group I (n = 9) - animals undergoing urethral segment resection with interposition of a patch of heterologous collagen matrix seeded with autologous smooth muscle cells; Group II (n = 9) - animals undergoing resection of a urethral segment with interposition of a decellularized heterologous collagen matrix patch. Two animals from each group were sacrificed on postoperative days seven, fourteen and twenty-eight; three animals from each group were sacrificed at the end of three postoperative months. At the end of the third month one animal from each group underwent urethroscopy for urethral integrity assessment and one animal from each group had its microcirculation image captured by a SDF device (Side-stream Dark Field - Microscan Analysis Software). One animal from each group in each euthanasia period underwent cystourethrography so as the urethra could be viewed at flow time. The matrices integration was assessed through histological examination using hematoxylin and eosin (H & E), Masson trichrome (MT), Picrosirius red and Von Willebrand staining. In a blind study with two pathologists all the slides were studied. RESULTS: The matrices whether seeded or not with autologous muscle cells were able to restore the architecture of the urethra, but were eliminated from the first week on, before incorporation. Microcirculation of the neourethra, at the end of the third month, showed the same characteristics as a normal urethra in both groups of animals. CONCLUSION: Natural heterologous matrices implanted in the urethra as onlay graft were not incorporated into its walls but were able to fully restore the cell architecture of the organ, regardless of being seeded or not with autologous muscle cells.

Integration of collagen matrices into the urethra when implanted as onlay graft

Objective To assess the integration of decellularized heterologous collagen matrices into the urethra, when implanted with no cells or when seeded with autologous smooth muscle cells. Materials and Methods Eighteen New Zealand rabbits were randomly assigned to two groups: Group I (n = 9) - animals undergoing urethral segment resection with interposition of a patch of heterologous collagen matrix seeded with autologous smooth muscle cells; Group II (n = 9) - animals undergoing resection of a urethral segment with interposition of a decellularized heterologous collagen matrix patch. Two animals from each group were sacrificed on postoperative days seven, fourteen and twenty-eight; three animals from each group were sacrificed at the end of three postoperative months. At the end of the third month one animal from each group underwent urethroscopy for urethral integrity assessment and one animal from each group had its microcirculation image captured by a SDF device (Side-stream Dark Field - Microscan Analysis Software). One animal from each group in each euthanasia period underwent cystourethrography so as the urethra could be viewed at flow time. The matrices integration was assessed through histological examination using hematoxylin and eosin (H&E), Masson trichrome (MT), Picrosirius red and Von Willebrand staining. In a blind study with two pathologists all the slides were studied. Results The matrices whether seeded or not with autologous muscle cells were able to restore the architecture of the urethra, but were eliminated from the first week on, before incorporation. Microcirculation of the neourethra, at the end of the third month, showed the same characteristics as a normal urethra in both groups of animals. Conclusion Natural heterologous matrices implanted in the urethra as onlay graft were not incorporated into its walls but were able to fully restore the ...

Breast cancer tissue slices as a model for evaluation of response to rapamycin.

Rapamycin is a selective inhibitor of the mammalian target of rapamycin (mTOR), a regulator kinase that integrates growth factors signaling via the phosphoinositide-3-kinase pathway and that has emerged as a novel therapeutic modality in breast cancer (BC). We propose a pre-clinical "ex-vivo" personalized organotypic culture of BC that preserves the microenvironment to evaluate rapamycin-mediated gene expression changes. Freshly excised ductal invasive BC slices, 400 µm thick (n=30), were cultured in the presence or absence (control) of rapamycin (20 nM) for 24 h. Some slices were formalin-fixed for immunohistochemical determinations and some were processed for microarray analysis. Control slices in culture retained their tissue morphology and tissue viability (detected by BrdU uptake). The percentage of proliferating cells (assessed by Ki67) did not change up to 24 h of treatment. Immunohistochemical evaluation of p-AKT, p-mTOR, p-4EBP1 and p-S6K1 indicated that AKT/mTOR pathway activation was maintained during cultivation. For microarray analysis, slices were divided into two groups, according to the presence/absence of epidermal growth factor receptor-type 2 and analyzed separately. Limited overlap was seen among differentially expressed genes after treatment (P<0.01) in both groups suggesting different responses to rapamycin between these BC subtypes. Ontology analysis indicated that genes involved in biosynthetic processes were commonly reduced by rapamycin. Our network analysis suggested that concerted expression of these genes might distinguish controls from treated slices. Thus, breast carcinoma slices constitute a suitable physiological tool to evaluate the short-term effects of rapamycin on the gene profile of individual BC samples.

Expression of claudin, paxillin and FRA-1 in non-nodular breast lesions in association with microcalcifications.

CONTEXT AND OBJECTIVE: The possible role of adhesion molecules in early breast carcinogenesis has been shown in the literature. We aimed to analyze early adhesion imbalances in non-nodular breast lesions and their association with precursor lesions, in order to ascertain whether these alterations exist and contribute towards early carcinogenesis. DESIGN AND SETTING: Retrospective cross-sectional study based on medical records at a private radiological clinic in São Paulo, Brazil. METHODS: We retrospectively reviewed the medical records of all consecutive women attended between August 2006 and July 2007 who presented mammographic evidence of breast microcalcifications classified as Breast Imaging Reporting and Data System Atlas (BI-RADS) type 4. These women underwent stereotaxic biopsy. Clinical, radiological and pathological data were collected, and immunohistochemical assays searched for claudin, paxillin, FRA-1 and HER-2. RESULTS: Over this period, 127 patients were evaluated. Previous BI-RADS diagnoses showed that 69 cases were in category 4A, 47 in 4B and 11 in 4C. Morphological assessment showed benign entities in 86.5%. Most of the benign lesions showed preserved claudin expression, associated with paxillin (P < 0.001). Paxillin and HER-2 expressions were correlated. FRA-1 expression was also strongly associated with HER-2 expression (P < 0.001). CONCLUSIONS: Although already present in smaller amounts, imbalance of adhesion molecules is not necessarily prevalent in non-nodular breast lesions. Since FRA-1 expression reached statistically significant correlations with radiological and morphological diagnoses and HER-2 status, it may have a predictive role in this setting.

Expression of claudin, paxillin and FRA-1 in non-nodular breast lesions in association with microcalcifications / Expressão de claudina, paxilina e FRA-1 em lesões mamárias não nodulares associadas a microcalcificações

CONTEXT AND OBJECTIVE The possible role of adhesion molecules in early breast carcinogenesis has been shown in the literature. We aimed to analyze early adhesion imbalances in non-nodular breast lesions and their association with precursor lesions, in order to ascertain whether these alterations exist and contribute towards early carcinogenesis. DESIGN AND SETTING Retrospective cross-sectional study based on medical records at a private radiological clinic in São Paulo, Brazil. METHODS We retrospectively reviewed the medical records of all consecutive women attended between August 2006 and July 2007 who presented mammographic evidence of breast microcalcifications classified as Breast Imaging Reporting and Data System Atlas (BI-RADS) type 4. These women underwent stereotaxic biopsy. Clinical, radiological and pathological data were collected, and immunohistochemical assays searched for claudin, paxillin, FRA-1 and HER-2. RESULTS Over this period, 127 patients were evaluated. Previous BI-RADS diagnoses showed that 69 cases were in category 4A, 47 in 4B and 11 in 4C. Morphological assessment showed benign entities in 86.5%. Most of the benign lesions showed preserved claudin expression, associated with paxillin (P < 0.001). Paxillin and HER-2 expressions were correlated. FRA-1 expression was also strongly associated with HER-2 expression (P < 0.001). CONCLUSIONS Although already present in smaller amounts, imbalance of adhesion molecules is not necessarily prevalent in non-nodular breast lesions. Since FRA-1 expression reached statistically significant correlations with radiological and morphological diagnoses and HER-2 status, it may have a predictive role in this setting. .

CONTEXTO E OBJETIVO A literatura tem mostrado a importância de moléculas de adesão na carcinogênese precoce de mama. Objetivamos analisar desequilíbrios precoces de adesão em lesões não nodulares da mama e associação com lesões precursoras, a fim de verificar se essas alterações existem e contribuem com a carcinogênese. TIPO DE ESTUDO E LOCAL Estudo retrospectivo baseado em prontuários médicos, numa clínica radiológica privada em São Paulo, Brasil. MÉTODOS Revisamos retrospectivamente prontuários de todas as mulheres consecutivamente atendidas com evidência mamográfica de microcalcificações mamárias, classificadas como tipo 4 do Breast Imaging Reporting and Data System Atlas (BI-RADS) entre agosto de 2006 e julho de 2007. Elas foram submetidas a biópsia estereotáxica. Dados clínicos, radiológicos e histopatológicos foram coletados e ensaios de imunoistoquímica procuraram por claudina, paxilina, HER-2 e FRA-1. RESULTADOS No período, 127 pacientes foram avaliadas. Diagnósticos de BI-RADS anteriores tinham 69 casos na categoria 4A, 47 em 4B, e 11 em 4C. A avaliação morfológica mostrou entidades benignas em 86,5%. A maioria das lesões benignas mostrou expressão preservada de claudina, associada a paxilina (P < 0,001). Expressões de paxilina e HER-2 foram correlacionadas. Expressão de FRA-1 associou-se à de HER-2 (P < 0,001). CONCLUSÕES Embora já presente em menor quantidade, o desequilíbrio de moléculas de adesão não é necessariamente prevalente em lesões mamárias nodulares e talvez a expressão de FRA-1 possa ter um papel preditivo neste cenário, uma vez que atingiu correlações ...

Analysis of CD83 antigen expression in human breast fibroadenoma and adjacent tissue / Análise da expressão do antígeno CD83 no fibroadenoma mamário humano e no tecido adjacente

CONTEXT AND OBJECTIVE: Dendritic cell maturation is considered essential for starting an immune response. The CD83 antigen is an important marker of dendritic cell maturation. The objectives here were to analyze CD83 antigen expression in human breast fibroadenoma and breast tissue adjacent to the lesion and to identify clinical factors that might influence this expression. DESIGN AND SETTING: This was a retrospective study at a public university hospital, in which 29 histopathological samples of breast fibroadenoma and adjacent breast tissue, from 28 women of reproductive age, were analyzed. METHODS: The immunohistochemistry method was used to analyze the cell expression of the antigen. The antigen expression in the cells was evaluated by means of random manual counting using an optical microscope. RESULTS: Positive expression of the CD83 antigen in the epithelial cells of the fibroadenoma (365.52; standard deviation ± 133.13) in relation to the adjacent breast tissue cells (189.59; standard deviation ± 140.75) was statistically larger (P < 0.001). Several clinical features were analyzed, but only parity was shown to influence CD83 antigen expression in the adjacent breast tissue, such that positive expression was more evident in nulliparous women (P = 0.042). CONCLUSIONS: The expression of the CD83 antigen in the fibroadenoma was positive and greater than in the adjacent breast tissue. Positive expression of the antigen in the adjacent breast tissue was influenced by parity, and was significantly more evident in nulliparous women.

CONTEXTO E OBJETIVOS: A maturação da célula dendrítica é considerada essencial para o início da resposta imune. O antígeno CD83 é um importante marcador da maturação da célula dendrítica. Os objetivos são analisar a expressão do antígeno CD83 no fibroadenoma mamário humano e no tecido mamário adjacente à lesão e identificar fatores clínicos que possam influenciar esta expressão. TIPO DE ESTUDO E LOCAL: Este é um estudo retrospectivo, realizado em um hospital público universitário, onde 29 amostras histopatológicas de fibroadenomas de mamas e de tecidos mamários adjacentes, de 28 mulheres em idade reprodutiva, foram analisados. MÉTODOS: O método de imunoistoquímica foi utilizado na análise da expressão celular do antígeno. A expressão do antígeno nas células foi avaliada por contagem aleatória e manual utilizando-se microcópio de luz. RESULTADOS: A expressão positiva do antígeno CD83 nas células epiteliais dos fibroadenomas (365,52; desvio padrão ± 133,13) em relação às células do tecido mamário adjacente (189,59; desvio padrão ± 140,75) foi estatisticamente superior (P < 0,001). Vários aspectos clínicos foram analisados, porém, a paridade se mostrou influente na expressão do antígeno CD83 no tecido mamário adjacente, onde a expressão positiva foi mais evidente nas mulheres nulíparas (P = 0,042). CONCLUSÕES: A expressão do antígeno CD83 foi positiva e mais expressiva no fibroadenoma do que no tecido mamário adjacente. A expressão positiva do antígeno no tecido mamário adjacente foi influenciada pela paridade, sendo significativamente mais evidente nas mulheres nulíparas.

Carcinoma mamário secretório em criança de sete anosde idade: relato de caso e revisão da literatura / Carcinoma mamário secretório em criança de sete anos de idade: relato de caso e revisão da literatura

O carcinoma secretório da mama é uma entidade extremamente rara, inicialmente descrita em crianças e adolescentes, mas um número relativamente frequente de casos foi relatado em adultos. É usualmente associado ao bom prognóstico. O presente caso refere-se a uma menina de sete anos de idade com crescimento tumoral progressivo na mama esquerda. O exame clínico identificou um nódulos ubareolar duro, móvel, indolor e margens regulares; macroscopicamente, o espécime cirúrgico mostrou nódulo com 1,3 cm de diâmetro e superfície lisa. O diagnóstico final de neoplasia maligna levou à mastectomia, e a avaliação intraoperatória do linfonodo sentinela foi negativa. O perfil imunoistoquímico incluiu coloração fortemente positiva para citoqueratinas (AE1/AE3) e TP53, expressão negativa para receptores de estrógeno e progesterona e proteína Her-2/neu; o antígeno de proliferação celualar Ki-67 (MIB-1) mostrou índice de marcação de 10%. Exames adicionais de acompanhamento não mostraram nenhuma evidência de metástases à distância, com desenvolvimento normal das características sexuais secundárias (pubarca, telarca direita e menarca, que ocorreu em julho de 2009). Sete anos mais tarde, a adolescente ainda está livre da doença e aguarda cirurgia plástica reconstrutiva da mama esquerda.

Secretory breast carcinoma is an extremely rare entity initially described in children and in adolescents, but a relatively frequent number of cases has been reported in adults. It is often associated with good prognosis. The present case refers to a seven-year-old girl with a progressively growing tumor in the left breast. Clinical examination identified a hard, mobile, non-painful,subareolar nodule with regular margins; further macroscopy of surgical specimen showed a smooth surface and 1.3 cm in diameter. The final diagnosis of malignancy led to mastectomy and intra-operatively negative sentinel lymph node assessment. Immunohistochemical profile included a strong positive stain for cytokeratins (AE1/AE3) and TP53, and estrogen-receptor and progesterone-receptor and Her-2/neu protein expression negative; and MIB-1 (Ki-67) labeling index was 10%. Further follow-up examinations have shown no evidence of distant metastases, with normal development of secondary sexual characteristics (pubarche, right thelarche, and menarche occurring in July, 2009). Seven years later, the teenager still does not have the disease and waits reconstructive plastic surgery of the left breast.

Expression of BCL-2 and bax proteins in breast cancer after in vivo tamoxifen administration / Expressão das proteínas BCL-2 e BAX no câncer de mama após administração in vivo de tamoxifeno

Apesar de apresentarem receptores estrogênicos nas células tumorais, algumas pacientes com câncer de mama não se beneficiam do tratamento com tamoxifeno, medicamento conhecido por interferir no ciclo celular e promover apoptose. Sabendo-se que as proteínas BCL-2 e BAx estão diretamente relacionadas a este processo, é importante compreender quais os caminhos que levam à morte celular que sofrem interferência do tamoxifeno. Amostras pareadas de câncer de mama foram obtidas antes e após tratamento de pacientes previamente, assim randomizadas: grupo controle e grupo de tratamento com tamoxifeno. O grupo de tratamento recebeu tamoxifeno (20 mg/dia) por 14 dias. A identificação imunoistoquímica da expressão de BAX e BCL-2 foi analizada de forma semiquantitativa, considerando o número de células coradas e a intensidade da coloração. Os escores de cada reação foram comparados pré e pós-tratamento, e também em relação ao grupo controle. Das 25 pacientes estudadas, considerando nenhuma exposição ao medicamento, foram encontradas 36 (9/25) e 72% (18/25) de casos positivos para BCL-2 e Bax, respectivamente. Este estudo não encontrou mudanças significativas sobre a expressão das proteínas BCL-2 e BAX, após 14 dias de tratamento com tamoxifeno (p>0,05), comparado ao grupo controle. A expressão das proteínas BCL-2 e BAX também não sofreu mudanças significativas após 14 dias de exposição ao tamoxifeno. Este é um dos poucos trabalhos prospectivos randomizados de estudo in vivo dos efeitos do tamoxifeno na apoptose.

Despite the presence of estrogen receptor in breast cancer cells, some patients do not show benefits on their treatmentwith tamoxifen, which is a medication that interferes on cell cycle promoting apoptosis. Since BCL-2 and BAX proteins are directly linked to this process, it is important to understand which pathways for cell death are related to and interfered by tamoxifen. Paired samples of breast cancer tumors, which were previously obtained before and after tamoxifen therapy were randomly divided in the Control and Treated Patients Groups. The Treated Group received tamoxifen for 14 days (20 mg/day). The immunohistochemical identification of BAX and BCL-2 expression was analyzed in a semi-quantitative scale consideraing the number of positive cells and intensity of staining. The scores of each reaction were compared pre and post-treatment and to the Control Group. Over the 25 patients studied, considering no exposure to the drug, there were 36 (9/25) and 72% (18/25) of positive cases for BCL-2 and BAX, respectively. This study showed no significant changes over BCL-2 and BAX proteins expressions after 14 days of tretament with tamoxifen (p>0,05) compared to the control patients. Expression of BCL-2 and BAX proteins did not suffer statistically significant changes after a 14-day exposure to tamoxifen. This is one of a few prospective randomized double-blind studies about in vivo effects of tamoxifen in apoptosis.

Analysis of CD83 antigen expression in human breast fibroadenoma and adjacent tissue.

CONTEXT AND OBJECTIVE: Dendritic cell maturation is considered essential for starting an immune response. The CD83 antigen is an important marker of dendritic cell maturation. The objectives here were to analyze CD83 antigen expression in human breast fibroadenoma and breast tissue adjacent to the lesion and to identify clinical factors that might influence this expression. DESIGN AND SETTING: This was a retrospective study at a public university hospital, in which 29 histopathological samples of breast fibroadenoma and adjacent breast tissue, from 28 women of reproductive age, were analyzed. METHODS: The immunohistochemistry method was used to analyze the cell expression of the antigen. The antigen expression in the cells was evaluated by means of random manual counting using an optical microscope. RESULTS: Positive expression of the CD83 antigen in the epithelial cells of the fibroadenoma (365.52; standard deviation ± 133.13) in relation to the adjacent breast tissue cells (189.59; standard deviation ± 140.75) was statistically larger (P < 0.001). Several clinical features were analyzed, but only parity was shown to influence CD83 antigen expression in the adjacent breast tissue, such that positive expression was more evident in nulliparous women (P = 0.042). CONCLUSIONS: The expression of the CD83 antigen in the fibroadenoma was positive and greater than in the adjacent breast tissue. Positive expression of the antigen in the adjacent breast tissue was influenced by parity, and was significantly more evident in nulliparous women.

Correlação entre as medidas antropométricas e o comprometimento linfonodal de pacientes pós-menopausadas com câncer de mama / Correlation between anthropometric measurements an lymph node metastases in postmenopausal women with breast cancer

Introdução: Diversos estudos epidemiológicos observaram que mulheres obesas na pós-menopausa apresentam aumento de risco e mortalidade do câncer de mama. Apesar de não haver consenso, alguns estudos demonstraram maior grau de comprometimento dos linfonodos axilares nessas pacientes, o que pode contribuir para o pior prognóstico da doença nesse grupo. Objetivo: Avaliar a correlação entre as medidas antropométricas e o grau de comprometimento linfonodal axilar em pacientes pós-menopausadas com câncer de mama hormônio-responsivo. Métodos: Estudo prospectivo analítico com 57 mulheres com carcinoma ductal invasivo no estágio II, na pós-menopausa, com receptores hormonais positivos (receptor estrogênico e/ou receptor de progesterona - RE/RP), tratadas nos hospitais São Paulo e Pérola Byington. Logo após o diagnóstico foram realizadas as medidas antropométricas (IMC, CA, CQ e RCQ) das pacientes e, após o tratamento cirúrgico e avaliação histopatológica dos linfonodos axilares, realizou-se o estudo estatístico. Resultados: Observou-se associação significativa entre o número de linfonodos acometidos e o sobrepeso (IMC> 25 kg/m2) (p = 0,0329). Cerca de 64% das pacientes com mais de três linfonodos acometidos apresentaram IMC> 25 kg/m2. Entretanto, não houve diferença estatística entre as medidas antropométricas e a positividade dos linfonodos axilares de forma global. Conclusão: O número de linfonodos axilares comprometidos foi maior em pacientes com índice de massa corpórea entre 25 e 30 kg/m2.

Introduction: Several epidemiological studies have shown an increased risk and mortality in breast cancer of obese postmenopausal women. The higher number of lymph node metastases in these patients could contribute to poor prognosis. Objective: To evaluate the correlation between the anthropometric measurements and lymph node metastases in postmenopausal women with breast cancer expressing hormone receptors (ER/PgR). Methods: Prospective study with 57 women with invasive ductal carcinoma, stage II and estrogen receptor and/or progesterone receptor (ER/PgR) positivity treated in São Paulo and Pérola Byington Hospital. Anthropometric datawere obtained after the diagnoses, and statistical analysis was done after surgery treatment and definitive pathology results of axillary lymph nodes dissection. Results: There was a significant association (p = 0.0329) between the number of axillary lymph node metastases and overweight (BMI > 25 kg/m2). Almost 64% of patients with more than three lymph node metastases had more than 25 kg/m2. However, there were no statistical significance between the correlation of anthropometric measurements and the global number of axillary lymph node metastases. Conclusion: The number of axillary lymph node metastases was higher in patients with body mass index between 25 and 30 kg/m2.

[Assessment of angiogenesis expression and its relationship with prognosis of colorectal cancer by conventional and computer-assisted histopathological image analysis]. / Investigação da expressão da angiogênese e sua relação com o prognóstico do câncer colorretal pela análise convencional e informatizada da imagem histopatológica.

PURPOSE: To quantify the degree of angiogenesis by conventional method (microvessel density, MVD) and computerized method (endothelial area, EA), and to evaluate their relationships with the prognosis of patients operated on for colorectal adenocarcinoma. METHODS: Tumoral angiogenesis was studied by means of an immunohistochemical technique, using CD 34, on 126 patients; to quantify the angiogenesis, MVD (defined as number of microvessels per mm(2)) and EA measurement (defined as the area occupied by EA in the microscope field). A computerized method, IMAGELab software was utilized to quantify endothelial area. RESULTS: The mean number of microvessels was 128.6 MV/mm(2) (SD = 44.5) and the mean EA was 4.3% (SD = 2.1). The Pearson method demonstrated a low correlation coefficient between MVD and EA (r = 0.429). No relationship between MVD and EA was observed with regard to relapse-free interval and overall survival. CONCLUSION: The histological analysis of angiogenesis expression in patients with colorectal adenocarcinoma can be performed either by computer-assisted image analysis of endothelial area or by conventional microvessels counting. Both methods did not show any significant relationship between these angiogenesis parameters with relapse-free interval and overall survival.

Assessment of angiogenesis expression and its relationship with prognosis of colorectal cancer by conventional and computer-assisted histopathological image analysis

PURPOSE: To quantify the degree of angiogenesis by conventional method (microvessel density, MVD) and computerized method (endothelial area, EA), and to evaluate their relationships with the prognosis of patients operated on for colorectal adenocarcinoma. METHODS: Tumoral angiogenesis was studied by means of an immunohistochemical technique, using CD 34, on 126 patients; to quantify the angiogenesis, MVD (defined as number of microvessels per mm²) and EA measurement (defined as the area occupied by EA in the microscope field). A computerized method, IMAGELab software was utilized to quantify endothelial area. RESULTS: The mean number of microvessels was 128.6 MV/mm² (SD = 44.5) and the mean EA was 4.3 percent (SD = 2.1). The Pearson method demonstrated a low correlation coefficient between MVD and EA (r = 0.429). No relationship between MVD and EA was observed with regard to relapse-free interval and overall survival. CONCLUSION: The histological analysis of angiogenesis expression in patients with colorectal adenocarcinoma can be performed either by computer-assisted image analysis of endothelial area or by conventional microvessels counting. Both methods did not show any significant relationship between these angiogenesis parameters with relapse-free interval and overall survival.

OBJETIVO: Quantificar a intensidade da angiogênese pelo método convencional (densidade microvasal, DMV) e pelo método informatizado, área endothelial (AE) e avaliar a sua correlação com o prognóstico de doentes operados por adenocarcinoma colorretal. MÉTODOS: A angiogênese tumoral foi investigada por meio de técnica imuno-histoquímica, utilizando-se CD-34, em 126 doentes; para quantificar a angiogênese, a microdensidade vascular, definida como o número de microvasos por mm² e a medida da área endotelial, definida como a área ocupada pelo endotélio vascular identificada no campo microscópico foram empregadas. Um programa computadorizado, o IMAGELab foi empregado para a quantificação da área endothelial. RESULTADOS: A media do número de microvasos foi de 128,6 MV/mm ² (d esvio padrão de 44,5). O método do coeficiente de Pearson demonstrou uma baixa correlação entre a DMV e a AE (r=0,429). Nenhuma correlação entre a DMV e AE com o intervalo livre de doença e tempo global de sobrevida foi observada. CONCLUSÃO: A análise histológica da expressão angiogênica em doentes com adenocarcinoma colorretal pode ser realizada tanto da forma informatizada, na quantificação da área endotelial, como pela convencional, na contagem dos microvasos. Ambos métodos não demonstraram relação estatisticamente significante entre estes parâmetros de angiogênese e o intervalo livre de doença e a sobrevida global.

The composition and behavior of capsules around smooth and textured breast implants in pigs.

BACKGROUND: The surface of the implant is one of the many factors often associated with the occurrence of capsular contracture, the etiopathogeny of which remains unclear. The purpose of this study was to analyze the behavior of capsular contracture by means of applanation tonometry and histology using a midsized animal model. METHODS: Silicone breast implants were implanted into 33 pigs and observed at 30, 60, 180, and 270 postoperative days. RESULTS: Capsular contracture in smooth implants showed significantly greater pressure values of tonometry, and the smooth implant capsule was significantly thicker than the textured implant capsule. Both pressure and thickness of the capsules increased at each period. The collagenous layer did not show any difference considering the periods of time in which the total thickness was analyzed; on the other hand, the increase in total capsular thickness occurred by thickening of the noncollagenous layer in both smooth and textured implants. Taking into consideration both kinds of implants, histomorphometric analysis showed that thin fibers were replaced by thick fibers in later postoperatives periods (180 and 270 days). CONCLUSIONS: The greater incidence of capsular contracture in smooth implants was correlated with the progressive increase in total capsule thickness, due to a higher concentration of collagenous fibers, when compared with textured implants (p = 0.011; mean difference, 6.61), and a higher concentration of thick fibers (p = 0.034; average, >5.51 percentage points per field of thick fibers than the textured implants in all periods). Pigs are good animal models for studying the healing process after breast augmentation with implants.