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Vasculogenic mimicry (VM) is the ability of highly aggressive cancer cells to form fluid-conducting channels that facilitate the nutrition and metastasis of cancer cells. Considering the importance of VM in the prognosis of canine mammary gland tumours, this study aimed to investigate global gene expression in two canine mammary carcinoma cell cultures associated with the capacity for VM in vitro. The cell lines were subjected to an in-vitro assay to form VM channels (3D culture). Each cell line was then used in 2D conditions as controls and we compared the global gene expression with that of the 3D cultures. A total of 1,217 differentially expressed genes (DEGs) (P <0.05, fold change >2.0 or <2.0) were observed in 3D conditions compared with 2D culture in the UNESP-CM9 cell line, of which 677 were upregulated genes and 540 were downregulated. In contrast, the UNESP-CM60 cell line had only one upregulated and two downregulated genes. Overall, we identified several genes and pathways involved in the development of VM and these molecular data will be useful for future studies aimed at identifying diagnostic and therapeutic targets for VM in canine mammary carcinoma.

Assuntos

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Rapamycin is an antifungal drug with antitumor activity and acts inhibiting the mTOR complex. Due to drug antitumor potential, the aim of this study was to evaluate its effect on a preclinical model of primary mammary gland tumors and their metastases from female dogs. Four cell lines from our cell bank, two from primary canine mammary tumors (UNESP-CM1, UNESP-CM60) and two metastases (UNESP-MM1, and UNESP-MM4) were cultured in vitro and investigated for rapamycin IC50. Then, cell lines were treated with rapamycin IC50 dose and mRNA and protein were extracted in treated and non-treated cells to perform AKT, mTOR, PTEN and 4EBP1 gene expression and global proteomics by mass spectrometry. MTT assay demonstrated rapamycin IC50 dose for all different tumor cells between 2 and 10 µM. RT-qPCR from cultured cells, control versus treated group and primary tumor cells versus metastatic tumor cells, did not shown statistical differences. In proteomics were found 273 proteins in all groups, and after data normalization 49 and 92 proteins were used for statistical analysis for comparisons between control versus rapamycin treatment groups, and metastasis versus primary tumor versus metastasis rapamycin versus primary tumor rapamycin, respectively. Considering the two statistical analysis, four proteins, phosphoglycerate mutase, malate dehydrogenase, l-lactate dehydrogenase and nucleolin were found in decreased abundance in the rapamycin group and they are related with cellular metabolic processes and enhanced tumor malignant behavior. Two proteins, dihydrolipoamide dehydrogenase and superoxide dismutase, also related with metabolic processes, were found in higher abundance in rapamycin group and are associated with apoptosis. The results suggested that rapamycin was able to inhibit cell growth of mammary gland tumor and metastatic tumors cells in vitro, however, concentrations needed to reach the IC50 were higher when compared to other studies.

Assuntos

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Canine prostate cancer (PC) presents a poor antitumor response, usually late diagnosis and prognosis. Toceranib phosphate (TP) is a nonspecific inhibitor of receptor tyrosine kinases (RTKs), including vascular endothelial growth factor receptor (VEGFR), platelet-derived growth factor receptor (PDGFR), and c-KIT. This study aimed to evaluate VEGFR2, PDGFR-ß, and c-KIT protein expression in two established canine PC cell lines (PC1 and PC2) and the transcriptome profile of the cells after treatment with TP. Immunofluorescence (IF) analysis revealed VEGFR2 and PDGFR-ß protein expression and the absence of c-KIT protein expression in both cell lines. After TP treatment, only the viability of PC1 cells decreased in a dose-dependent manner. Transcriptome and enrichment analyses of treated PC1 cells revealed 181 upregulated genes, which were related to decreased angiogenesis and cell proliferation. In addition, we found upregulated PDGFR-A, PDGFR-ß, and PDGF-D expression in PC1 cells, and the upregulation of PDGFR-ß was also observed in treated PC1 cells by qPCR. PC2 cells had fewer protein-protein interactions (PPIs), with 18 upregulated and 22 downregulated genes; the upregulated genes were involved in the regulation of parallel pathways and mechanisms related to proliferation, which could be associated with the resistance observed after treatment. The canine PC1 cell line but not the PC2 cell line showed decreased viability after treatment with TP, although both cell lines expressed PDGFR and VEGFR receptors. Further studies could explain the mechanism of resistance in PC2 cells and provide a basis for personalized treatment for dogs with PC.

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Soft tissue sarcomas are a heterogeneous group of malignant neoplasms with different morphological patterns of mesenchymal lineage. This type of neoplasm is most commonly found in the subcutaneous tissue but is rare in solid organs, such as the liver and kidneys. This paper describes the main anatomopathological alterations in the liver of a Wistar rat(Rattus norvegicus) with soft tissue sarcoma. A two-year-old male pet Wistar rat was referred to the Laboratory of Veterinary Pathology at the Federal University of Fronteira Sul. The owner reported apathy and the animal died during physical examination. At necropsy, 10 ml of a reddish liquid was found in the thoracic cavity. The left lateral liver lobec ontained a 5-cm mass of heterogeneous surface composed of whitish parenchyma and red multifocal lesions extending to the surface of the liver. Moreover, whitish dotted spots intercalated with dark and more friable spots were found in the whole left lateral liver lobe. Histopathological evaluation of the nodule revealed the formation of spindle cells in parallel bundles with slightly eosinophilic cytoplasm, elongated nucleus, hyperchromatic to granular chromatin, and inconspicuous nucleolus. Extracellular matrix and mineralization were also observed. An area with proliferation of spindle cells with elongated cytoplasm, round to oval nuclei, sometimes hyperchromatic, consistent with cells found in the liver was noted in the mesenteric lymph node and omental node. Masson Trichrome staining revealed tumor cells stained in blue. Immunohistochemistry was performed and revealed positive staining for vimentin and negative for pan-cytokeratin, S100,desmin and factor VIII. Thus, the histological, histochemical and immunohistochemical evaluations suggested hepatic fibrosarcoma. This report showed the histological and immunohistochemical findings of a poorly described tumor in a Wister rat in veterinary literature.

Assuntos

RESUMO

Soft tissue sarcomas are a heterogeneous group of malignant neoplasms with different morphological patterns of mesenchymal lineage. This type of neoplasm is most commonly found in the subcutaneous tissue but is rare in solid organs, such as the liver and kidneys. This paper describes the main anatomopathological alterations in the liver of a Wistar rat(Rattus norvegicus) with soft tissue sarcoma. A two-year-old male pet Wistar rat was referred to the Laboratory of Veterinary Pathology at the Federal University of Fronteira Sul. The owner reported apathy and the animal died during physical examination. At necropsy, 10 ml of a reddish liquid was found in the thoracic cavity. The left lateral liver lobec ontained a 5-cm mass of heterogeneous surface composed of whitish parenchyma and red multifocal lesions extending to the surface of the liver. Moreover, whitish dotted spots intercalated with dark and more friable spots were found in the whole left lateral liver lobe. Histopathological evaluation of the nodule revealed the formation of spindle cells in parallel bundles with slightly eosinophilic cytoplasm, elongated nucleus, hyperchromatic to granular chromatin, and inconspicuous nucleolus. Extracellular matrix and mineralization were also observed. An area with proliferation of spindle cells with elongated cytoplasm, round to oval nuclei, sometimes hyperchromatic, consistent with cells found in the liver was noted in the mesenteric lymph node and omental node. Masson Trichrome staining revealed tumor cells stained in blue. Immunohistochemistry was performed and revealed positive staining for vimentin and negative for pan-cytokeratin, S100,desmin and factor VIII. Thus, the histological, histochemical and immunohistochemical evaluations suggested hepatic fibrosarcoma. This report showed the histological and immunohistochemical findings of a poorly described tumor in a Wister rat in veterinary literature.(AU)

Assuntos

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Canine cutaneous squamous cell carcinoma (cSCC) is the most common skin cancer in dogs, and, due to its low metastatic rate, local treatments, such as electrochemotherapy (ECT), promote disease control or even complete remission (CR). This study aimed to evaluate the gene and protein expression of Bcl-2 and Bcl-2 associated X protein (BAX), the proliferative index and clinical parameters in dogs with cSCC subjected to ECT. A prospective nonrandomized clinical study was performed using dogs with naturally occurring cSCC that was treated with ECT. Eighteen lesions from 11 dogs were selected. The tumor size at day 0 (D0) had no impact on survival or prognosis (P > 0.05). Tumor samples had a lower proliferative index after ECT (D21) than before ECT (P = 0.031). The survival of subjects with Ki67 values lower and higher than the Ki67 median value were not significantly different (P > 0.05). Regarding apoptotic markers, there were no significant differences in the gene and protein expression levels of BAX or Bcl-2 at D0 and D21 (P > 0.05) or in the overall survival of subjects with different levels of apoptotic markers. In conclusion, there was no change in BAX or Bcl-2 gene and protein expression in response to ECT at the time points evaluated, but ECT was able to reduce tumor volume and cellular proliferation in cSCC.

Assuntos

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In veterinary medicine, primary mammary hemangiosarcoma is a very rare disease and there was no previous report describing the disease in dogs. Herein, we describe necropsy findings of a female dog, which presented a diffuse mammary hemangiosarcoma affecting the mammary gland. A diffuse irregular plaque was found in all mammary gland, involving the whole mammary gland tissue. The histopathological evaluation revealed neoplastic cells with an ovoid to spindle shaped basophilic and numerous atypical neoplastic tumor cells forming vascular structures. There was no glandular proliferation and no changes in the superficial dermis and no neoplastic emboli in the lymph vessels. Immunohistochemistry for vimentin, pan-cytokeratin, CK8/18 and CD31 was conducted. Positive expression was found to the pan-cytokeratin and CK8/18 by remaining epithelial cells confirmed the luminal mammary origin. Vimentin and CD31 positive expression by neoplastic cells confirmed the endothelial origin of the neoplasia. The histopathological and immunohistochemical findings supported the diagnosis of a primary mammary hemangiosarcoma.

Assuntos

RESUMO

In veterinary medicine, primary mammary hemangiosarcoma is a very rare disease and there was no previous report describing the disease in dogs. Herein, we describe necropsy findings of a female dog, which presented a diffuse mammary hemangiosarcoma affecting the mammary gland. A diffuse irregular plaque was found in all mammary gland, involving the whole mammary gland tissue. The histopathological evaluation revealed neoplastic cells with an ovoid to spindle shaped basophilic and numerous atypical neoplastic tumor cells forming vascular structures. There was no glandular proliferation and no changes in the superficial dermis and no neoplastic emboli in the lymph vessels. Immunohistochemistry for vimentin, pan-cytokeratin, CK8/18 and CD31 was conducted. Positive expression was found to the pan-cytokeratin and CK8/18 by remaining epithelial cells confirmed the luminal mammary origin. Vimentin and CD31 positive expression by neoplastic cells confirmed the endothelial origin of the neoplasia. The histopathological and immunohistochemical findings supported the diagnosis of a primary mammary hemangiosarcoma.(AU)

Assuntos