Liliane Cunha Campos

Histopathological and immunohistochemical assessment of invasive micropapillary mammary carcinoma in dogs: A retrospective study

Invasive micropapillary carcinoma (IMPC) of the mammary gland, despite its rare occurrence in humans and dogs, is an important neoplasm due to its aggressive behaviour. The aim of this study was to evaluate the clinicopathological and immunophenotypical characteristics of IMPC and to determine the overall survival of dogs with this tumour. Of the selected cases, the majority had >3 cm neoplasms (15/19, 78.95%) and lymph node metastases (16/16, 100%), but only two cases (2/9, 22.2%) had distant metastases. The IMPCs were classified as either pure (15/22, 68.18%) or mixed (7/22, 31.82%) types. There was a predominance of moderate histological grade tumours (16 grade II) and the average overall survival was 120 days. Positive immunohistochemical staining for epithelial membrane antigen and negative staining for CD-31, p63 and cytokeratin (CK) AE1AE3 in cystic formations confirmed the micropapillary nature of these neoplasms. A proportion of cases exhibited positive epithelial staining for p63 (4/20, 20%) and CK34βE12 (20/22, 90.9%). Most cases were positive for oestrogen (19/20, 95%) and progesterone (19/20, 95%) receptors, but lacked HER-2 (16/22, 72.72%) and epidermal growth factor receptor (15/22, 68.18%) over-expression. The mean proliferation index was 14.8%. The findings demonstrate that, similar to humans, canine IMPCs behave aggressively with high rates of metastasis to regional lymph nodes and short overall survival times.

Combination therapy with carboplatin and thalidomide suppresses tumor growth and metastasis in 4T1 murine breast cancer model.

Carboplatin, efficient cytostatics for cancer therapy, could induce apoptosis and inhibit the growth of vascular endothelium in several tumor cell lines and xenograft models. It has been suggested that the antitumor effect of chemotherapy could be increased by combining it with an antiangiogenesis agent in anticancer strategy. The present study explored the potential to increase the antitumor effect of carboplatin by combining it with thalidomide in mouse 4T1 breast cancer models, and the underlining mechanism was investigated. The systemic administration of carboplatin and thalidomide significantly decreased tumor growth through increased tumor cell apoptosis compared with either control group. Collectively, these findings suggest that combined treatment has shown synergistic suppression in tumor progression according to the analysis. Furthermore, also was observed reduction in number of lung metastases as compared to isolated treatments and increased survival of the animals. The present study may be important in future exploration of the potential application of the combined approach in the treatment of breast cancer.

Quantitation of the Regional Lymph Node Metastatic Burden and Prognosis in Malignant Mammary Tumors of Dogs.

Background As in women, regional lymph node status impacts survival in dogs with malignant mammary tumors. However, few studies have evaluated regional lymph node metastases in dogs with malignant mammary gland tumors. Objectives To estimate overall survival based on the assessments of the lymph node status and the morphologic and morphometric features in female dogs with malignant mammary gland tumors. Materials and Methods In total, 178 lymph nodes from 97 female dogs were assessed and reviewed, and after confirmation by immunohistochemistry (IHC), 161 lymph nodes were selected for analysis of metastases. Animals were considered metastasis‐free (negative lymph nodes) only after IHC analysis for cytokeratin AE1/AE3. The number of positive lymph nodes, the number of metastatic foci, the maximum diameter and the area of metastasis were analyzed, and estimates of overall survival were made. Results Dogs with metastasis had lower mean survival than those with metastasis‐free regional lymph nodes, showing a direct relationship between the number of affected lymph nodes and shorter survival. However, histologic analysis of the lymph nodes identified lower survival rates in animals with macrometastases and isolated tumor cells, areas of metastasis >20.11 mm², and metastatic diameters >7.32 mm. Conclusion The identification of ≥1 lymph nodes positive for metastasis and morphometric characterization of lymphatic metastases indicate the prognostic relevance of lymph nodes status in dogs with mammary tumors.

Myeloid metaplasia in canine mixed mammary tumors: occurrence and characterization

Background: Mixed tumors are among the most frequent mammary neoplasms in female dogs. Some of these tumors present bone marrow associated with the newly formed osseous tissue, characteristic of myeloid metaplasia. Objective: To evaluate the occurrence of these lesions in a series of mixed tumors, and determine its histomorphological characteristics. Animals and methods: In total, 384 canine mammary mixed tumors from 289 animals have been reviewed. The lesions were classified according to the presence of osseous metaplasia associated with myeloid metaplasia or extramedullary hematopoiesis. Myeloid metaplasia characterization was determined from the morphological characteristics and organization of the cells and adjacent tissues. Cytoplasmic staining for CD31 and Factor VIII were used as a criterion to confirm the presence of blood vessels and megakaryocytes, respectively. Results: The 384 cases included 206 benign and 178 carcinomas in mixed tumors. Osseous metaplasia was present in 16.1% and calcified areas exclusively in 3.1% lesions. Among all osseous metaplasia, 33.9% presented some type of extramedullary hematopoiesis, of which 71.4% were classified as myeloid metaplasia and 28.6% as extramedullary hematopoiesis. Myeloid metaplasia cases consisted of 67% benign mixed tumors and 33% carcinomas in mixed tumors. CD31 and Factor VIII expression occurred in all myeloid metaplasia, confirming the presence of blood capillaries and megakaryocytes. Myeloid metaplasia was observed in 24% of mixed tumors containing osseous metaplasia and in 4% of all mixed tumors analyzed. Conclusion: Despite the low frequency of this lesion, additional studies are needed to understand the implications of myeloid metaplasia in canine mammary mixed tumors.

Prognostic significance of tissue and serum HER2 and MUC1 in canine mammary cancer

The aim of our study was to compare serum levels and protein tissue of human epidermal growth factor receptor–2 proto-oncogene (HER2) and mucin 1 (MUC1) using an antigen-capture enzyme-linked immunosorbent assay and immunohistochemistry (IHC) in canine mammary carcinomas and investigate how the 2 markers correlate with dogs with metastasis and without metastasis to a regional lymph node. Forty-eight female dogs were selected, including 14 with non-metastatic cancer, 14 with lymph node metastasis, and 20 healthy animals. Serum samples were collected from all the animals and tissues from 28 dogs with malignant mammary tumor with or without metastasis for evaluated HER2 and MUC1 expression. Tissue sample were evaluated for MUC1 and HER2 immunoexpression by IHC. The results showed measurable serum levels of MUC1 and HER2 in all groups. While serum MUC1 levels were significantly higher in animals with metastasis than the other 2 groups, no increase was observed in HER2 serum levels. The MUC1 IHC results showed that only membrane immunostaining was significantly different between the groups. Statistically, there was an association between immunostaining and the serum HER2 levels. Our results indicate that serum concentrations of HER2 and the IHC staining pattern for HER2 in primary tumor do not correlate with the presence of regional metastasis. However, increased concentrations of MUC1 in the serum of dogs with mammary cancer are associated with the presence of metastasis to regional lymph nodes. A membrane pattern of IHC staining for MUC1 in the primary tumor suggests that metastases to regional lymph node are present.

Versican expression in myoepithelial cells from carcinomas in canine mixed mammary tumors

The matrix of canine mixed mammary tumors (CMMTs) consists of proliferating spindle cells of possible myoepithelial origin, as well as myxomatous tissue, cartilage matrix and/or bone. Among the multiple components of this tumor extracellular matrix, versican probably plays a prominent role due to its importance in tumor progression, cell proliferation and differentiation. However, there are few data related to a possible association between versican expression and the state of myoepithelial cell differentiation in CMMTs. Using immunohistochemistry and histochemistry, the objective of this study was to evaluate the expression of versican, sulfated proteoglycans and mucopolysaccharides in myoepithelial cells at different stages of differentiation and to explore a potential relationship with p63 and α-smooth muscle actin (SMA) expression. A significant difference in versican expression was observed among the different stages of myoepithelial cell differentiation with an inverse correlation between versican and p63/SMA expression. These results suggest that at an early stage of proliferation, myoepithelial cells acquire a phenotype consistent with a role in chondrogenesis. Moreover, myoepithelial cells showed an affinity for safranin and periodic acid-Schiff staining at different stages of proliferation supporting the myoepithelial origin of spindle cells from CMMTs.

HER-2, EGFR, Cox-2 and Ki67 expression in lymph node metastasis of canine mammary carcinomas: Association with clinical-pathological parameters and overall survival.

Studies about canine mammary tumors based on single molecular markers probably cannot accurately account for the heterogeneity of this disease, and the investigation of multiple molecular alterations in primary tumors and their metastases, in conjunction, has assumed great importance for the understanding of mammary tumor progression. In the present study, we selected 54 primary mammary carcinomas with lymph node metastasis (T1,2,3N1M0), 29 primary mammary carcinomas without metastasis (T1,2,3N0M0), and 25 canine lymph nodes metastasis to evaluate the immunohistochemical expression of HER-2, EGFR, Cox-2 and Ki67 and its association with clinical-pathological parameters and overall survival. Our results found a concordance between the expression of HER-2 (K coefficient: 0.250), Cox-2 (K coefficient: 0.571), and Ki67 (K coefficient: 0.397) and a discordance between EGFR expression (K coefficient: -0.195) in primary mammary carcinomas and paired lymph node metastasis. Furthermore, a high Ki67 index (>24%), large tumor size and the presence of angiolymphatic invasion in canine primary mammary carcinoma with lymph node metastasis plus the presence of extracapsular extension in lymph nodes metastasis were also related to worse prognoses and shorter overall survival (P<0.05). In conclusion, our study demonstrates that primary mammary carcinomas with high expression of HER-2, Cox-2 and Ki67 also show high expression of these markers in paired lymph node metastasis. Moreover, the expression of these molecular markers in lymph nodes metastasis did not demonstrate a prognostic relevance.

Thalidomide promotes leukocytosis in mice inoculated with 4T1 mammary carcinoma

The aim of this study was to evaluate the therapy effect of thalidomide in the 4T1 murine mammary carcinoma. 4T1 cell suspension was injected into the posterior left flank of all animals to obtain solid tumors. Five days after inoculation, the treatment group was orally administered 150 mg/kg of thalidomide for seven days. Tumors were measured every 48 hours until the end of treatment. Whole blood was collected for hematology analysis. Our results suggest that thalidomide therapy increase the number of circulating leukocytes in the 4T1 murine mammary carcinoma, and this response is accompanied by a decrease in tumor growth.

Avaliação nutricional de pacientes com câncer de mama atendidas no Serviço de Mastologia do Hospital das Clínicas, Belo Horizonte (MG), Brasil

The scope of this article is to evaluate the nutritional status, body composition and tumor characteristics of 31 patients with breast cancer attended at the Mastology Service of Hospital das Clinicas of the Federal University of Minas Gerais. Dietary data were obtained from the 24-hour dietary recall in the pre-operative state and analyzed by the DietWin Professional 2008 Nutrition Software. Body composition was assessed by bioelectrical impedance analysis. The tumor characterization data were collected from medical records. The mean age of patients was 50.75 ± 14.34 years. Excess body weight was found in 58% and waist circumference greater than 80 cm in 64.5% of patients Excessive consumption of oils and sugars was observed in 90.3% and 83.8%, respectively. Most had low intakes of calcium and vitamins B6, B12 and A. The predominant diagnosis was type II histological grade invasive ductal carcinoma in stage II or III. The prevalence of overweight and inadequate dietary intake demonstrate the need for individualized nutritional guidance and monitoring to improve the prognosis and quality of the life of patients.

Canine low-grade intra-orbital myxosarcoma: case report

OBJECTIVE The aim of this study was to evaluate important clinical, morphological, histopathological, histochemical, and immunohistochemical characteristics in order to establish the diagnosis and prognosis of a low-grade intra-orbital myxosarcoma. ANIMAL STUDIED A mongrel dog presented a 2-year history of a neoplastic mass behind the right eye. RESULTS The neoplasm presented a mesenchymal spindle and stellate cell proliferation with an abundant myxoid matrix, moderate anisocariosis, and a low mitotic index. It stained positive for vimentin, moderately positive for periodic acid-Schiff, and negative for Gomori trichrome stain and α-smooth muscle actin. CONCLUSIONS One year following surgical excision, the patient remains disease free. The histological findings established a diagnosis of a rare canine intra-orbital low-grade myxosarcoma.