Comparação dos métodos de palpação retal, citologia, histologia e imunoistoquímica para o diagnóstico da hiperplasia prostática benigna no cão

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detecção e expressão dos genes supressores p53 E c-Myc em tumores palpebrais de cães

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo anátomo-clínico do linfonodo sentinela no câncer de mama

Pós-graduação em Ginecologia, Obstetrícia e Mastologia - FMB

Imunofenotipagem de lesões obtidas em carcinogênese quimicamente induzida por DMBA em glândulas salivares submandibulares e ratos (Rattus norvegicus)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Metástases de carcinoma de mama em cadelas: avaliação da técnica de linfonodo sentinela

Pós-graduação em Medicina Veterinária - FMVZ

Immunoexpression of S100A4 in canine skin melanomas and correlation with histopathological parameters

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Epithelial-mesenchymal transition occurs in preneoplastic and neoplastic lesions of canine prostate

Background: The epithelial-mesenchymal transition (EMT) is an essential process in the tumor progression and metastasis. In human prostate carcinoma (PCa), the upregulation of cytokeratin and E-cadherin and down-regulation of vimentin have been associated with aggressive phenotype and poor prognosis. Due to the importance of canine cancer model it was evaluated the immunoexpression of AE1/AE3, E-cadherin and vimentin in canine prostatic lesions. Patients and Methods: A total of 75 prostatic tissues formalin-fixed paraffin embedded from dogs was selected: 10 normal prostatic tissues, 20 benign prostatic hyperplasia (BPH), 25 proliferative inflammatory atrophy (PIA) and 20 PCa. AE1/AE3 was detected with a monoclonal antibody (Invitrogen, 180132) at a 1:300 dilution, applied for 45 min at room temperature (RT). The antibody against Vimentin (V9, Invitrogen) and E-cadherin (NCH-38, Dako cytomatiomn) were monoclonal mouse antibodies, used at a 1:300 and 1:200, respectively, for 45 min at RT. The immunolabelling was performed by a polymer method (Histofine, Nichirei Biosciences,). A negative control was performed for all antibodies by omitting the primary antibody and substituting with Tris-buffered saline. The percentage of C-MYC, E-cadherin, and p63- positive cells per lesion was evaluated according to Prowatke et al. (2007). The samples were scored separately according to staining intensity and graded semi-quantitatively as negative, weakly positive, moderately positive, and strongly positive. The score was done in one 400 magnification field, considering only the lesion, since this was done in a TMA core of 1 mm. For statistical analyses, the immunostaining classifications were reduced to two categories: negative and positive. The negative category included negative and weakly positive staining. Chi-square or Fisher exact test was used to determine the association between the categorical variables. Results: All prostatic normal and BPH tissue were positive for cytokeratin, E-cadherin and negative for vimentin. Similarly, all PIA samples were positive for AE1/AE3. From those samples, 48% (12/25) were also positive for vimentin. 55% of PCa (11/25) was positive for vimentin and among these samples 75% (6/11) was also positive for AE1/AE3 and 45% (5/11) was negative for AE1/AE3. PIA and PCa presented a higher number of vimentin positive cells when compared with normal tissue (p=0.032). E-cadherin expression had no statistical difference among diagnosis groups, but we found a higher number of positive cases, with more than 51% of positive immunostaining in BPH and PIA (81.25% and 78.60% of the cases, respectively) than in PCa (55.55%). Conclusion: The carcinogenesis process regarding prostatic epithelial cells in dogs showed higher vimentin protein expression associated with concomitant loss of the cytokeratin and E-cadherin, similar in humans.

Primary squamous cell carcinoma of the thyroid diagnosed as anaplastic carcinoma: failure in fine-needle aspiration cytology?

A case of primary squamous-cell carcinoma (SCC) of the thyroid which had been initially diagnosed as an anaplastic carcinoma (ATC) is described: female, 73 years old, with a fast-growing cervical nodule on the left side and hoarseness for 3 months. Ultrasonography showed a 4.5 cm solid nodule. FNA was compatible with poorly differentiated carcinoma with immunoreactivity for AE1/AE3, EMA. Thyroidectomy was performed. Histopathological examination showed a nonencapsulated tumor. Immunohistochemistry disclosed positivity for AE1/AE3, p53,p63, and Ki67. The diagnosis was ATC. A second opinion reported tumor consisting of squamous cells, with intense inflammatory infiltrate both in tumor and in the adjacent thyroid, with final diagnosis of SCC, associated with Hashimoto thyroiditis. No other primary focus of SCC was found. Patient has shown a 48-month survival period. Clinically, primary SCCs of the thyroid and ATCs are similar. The distinction is often difficult particularly when based on the cytological analysis of FNA material.

Análise das células tronco tumorais em neoplasias mamárias de cães

Mammary tumors are the most frequent cancers in dogs, representing about 50% of tumors, and have a higher incidence in females of middle aged and elderly. These tumors have been used as a model for breast cancer in women due to several common characteristics such as histological and immunohistochemical similarities. In the last decade, studies based on molecular profiles of breast cancer, made possible the identification of some neoplastic cells with characteristics of stem cells - cancer stem cells (CSC). One of the putative molecules of CSCs is CD44. Recent studies have established a crucial link between the epithelial-mesenchymal transition (EMT) and the acquisition of molecular and functional properties of stem cells. For that reason we analyzed the expression of proteins CD44, Cytokeratins AE1/AE3 and Vimentin, in dogs mammary tumors, to investigate the potencial for CSC markers, and its relation with the EMT using immunohistochemistry in paraffin embedded tissues making use of techniques such as Tissue MicroArrays (TMA). Immunostaining of cytokeratin had no significant difference between benign and malignant tumors (p ≥ 0,05), being more intense in malignant tumors. However vimentina showed higher staining intensity in benign tumors, but with no significant difference (p ≤ 0,05). The expression of CD44 was higher in malignant tumors that have greater proliferative and metastatic potencial, however its relation with EMT was not detected in the analyzed tumors. The techniques applied for the TMAs were efficient and can be used in routine and later researches.

Malignant epitheliod hemangioendothelioma of the thyroid: a case report

Primary malignant epithelioid hemangioendothelioma (MEH) of the thyroid is a rare neoplasia with only a few cases reported in the literature. We report a 75 year old man, who presented with a substernal goiter and compressive symptoms. Ultrasonographic evaluation revealed a hypoechogenic nodule in the left lobe, measuring 4.1 cm in maximum diameter, and associated gross calcifications. Fine needle aspiration yielded hemorrhagic material. A left thyroid lobectomy and isthmusectomy was performed. The surgical specimen contained a malignant epithelioid hemangioendothelioma measuring 6 x 4 x 3 cm that had infiltrated about 50% of the thyroidal parenchyma, and surrounded a necrotic nodule. Immunohistochemistry results corroborated the histopathological findings; staining was positive for AE1/AE3, CD31, CD34, factor VIII-related antigen, and Ki-67 expression. Because of the patient’s comorbidities, surgical complementation was not undertaken and he has been undergoing conservative treatment.

Proposal for a 10-high-power-fields scoring method for the assessment of tumor budding in colorectal cancer

Although tumor budding is linked to adverse prognosis in colorectal cancer, it remains largely unreported in daily diagnostic work due to the absence of a standardized scoring method. Our aim was to assess the inter-observer agreement of a novel 10-high-power-fields method for assessment of tumor budding at the invasive front and to confirm the prognostic value of tumor budding in our setting of colorectal cancers. Whole tissue sections of 215 colorectal cancers with full clinico-pathological and follow-up information were stained with cytokeratin AE1/AE3 antibody. Presence of buds was scored across 10-high-power fields at the invasive front by two pathologists and two additional observers were asked to score 50 cases of tumor budding randomly selected from the larger cohort. The measurements were correlated to the patient and tumor characteristics. Inter-observer agreement and correlation between observers' scores were excellent (P<0.0001; intraclass correlation coefficient=0.96). A test subgroup of 65 patients (30%) was used to define a valid cutoff score for high-grade tumor budding and the remaining 70% of the patients were entered into the analysis. High-grade budding was defined as an average of ≥10 buds across 10-high-power fields. High-grade budding was associated with a higher tumor grade (P<0.0001), higher TNM stage (P=0.0003), vascular invasion (P<0.0001), infiltrating tumor border configuration (P<0.0001) and reduced survival (P<0.0001). Multivariate analysis confirmed its independent prognostic effect (P=0.007) when adjusting for TNM stage and adjuvant therapy. Using 10-high-power fields for evaluating tumor budding has independent prognostic value and shows excellent inter-observer agreement. Like the BRE and Gleason scores in breast and prostate cancers, respectively, tumor budding could be a basis for a prognostic score in colorectal cancer.

Intratumoral budding is preoperative biopsies predicts local and distant metastasis in colorectal cancer patients

Objective: In 2011, the term “intratumoral budding, ITB” was used to describe the presence of tumor buds within the main tumor body and was correlated to worse clinical outcome in colorectal cancer patients. Here, we further elucidate the potential clinical role of ITB in pre-operative biopsies using pan-cytokeratin stained tissues and a quantitative scoring system. Method: 139 pre-operative biopsies from patients with colorectal cancer underwent immunohistochemistry for pancytokeratin (AE1/AE3). ITB were counted in the area of densest budding (40×) and classified as high-grade when >10 buds/HPF were observed based on receiver operating characteristic (ROC) curve analysis. Results: High-grade ITB occurred in 26.6 % of cases and was associated with right-sided tumor location (p=0.0356), more advanced pT (p=0.0198) and pN (p<0.0001) classifications, distant metastasis (p=0.0164), higher tumor grade (p=0.0037) and lymphatic invasion (p=0.0445). The specificity and positive predictive value for lymph node metastasis was 86.7 % and 75.6 %, respectively. Disease-free survival was significantly worse in patients with high-grade ITB (5-year survival=25 %) in comparison to patients with low-grade ITB (5-year survival=55 %) (p=0.0157). Conclusion: The assessment of ITB in pre-operative biopsies is predictive of local and distant metastasis in corresponding resections and should be considered in daily management of colorectal cancer patients.

Intratumoural budding (ITB) in preoperative biopsies predicts the presence of lymph node and distant metastases in colon and rectal cancer patients

Background:In colorectal cancer (CRC), tumour budding at the invasion front is associated with lymph node (LN) and distant metastasis. Interestingly, tumour budding can also be detected in biopsies (intratumoural budding; ITB) and may have similar clinical importance. Here we investigate whether ITB in preoperative CRC biopsies can be translated into daily diagnostic practice.Methods:Preoperative biopsies from 133 CRC patients (no neoadjuvant therapy) underwent immunohistochemistry for pan-cytokeratin marker AE1/AE3. Across all biopsies for each patient, the densest region of buds at × 40 (high-power field; HPF) was identified and buds were counted.Results:A greater number of tumour buds in the biopsy was associated with pT stage (P=0.0143), LN metastasis (P=0.0007), lymphatic (P=0.0065) and venous vessel invasion (P=0.0318) and distant metastasis (cM1) (P=0.0013). Using logistic regression, a 'scale' was developed to estimate the probability of LN and distant metastasis using the number of tumour buds (e.g. 10 buds per HPF: 64% chance of LN metastasis; 30 buds per HPF: 86% chance). Inter-observer agreement for ITB was excellent (intraclass correlation coefficient: 0.813).Conclusion:Tumour budding can be assessed in the preoperative biopsy of CRC patients. It is practical, reproducible and predictive of LN and distant metastasis. Intratumoural budding qualifies for further investigation in the prospective setting.