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Canine Testicular Tumours: Correlation of Mitotic Index with Ki67 Expression
150:1, 2014
ESVP/ECVP Proceedings 2013
111
PRIMARY ANGIOCENTRIC/ANGIOINVASIVE T-CELL LYMPHOMA OF THE TYMPANIC BULLA IN A FeLVPOSITIVE CAT S.F. Santagostino, C.M. Mortellaro, A. Forlani, G. Ghisleni and P. Roccabianca DIVET-Dipartimento di Scienze Veterinarie e Sanita Pubblica, University of Milano, Italy Introduction: Reports of primary middle ear lymphoma are rare in cats and their extension to internal organs is exceptional. A 5-yearold, neutered female, FeLV-positive domestic shorthair cat was referred for stertor, dyspnoea and head-tilt. CT scan revealed soft tissue opacity inside the right tympanic bulla with bone lysis and concurrent nasopharyngeal and intracranial invasion. Materials and Methods: Endoscopic-guided biopsy samples were collected for histology and immunohistochemistry. A full necropsy examination was performed. Results: Grossly, the lesion was poorly demarcated, white and soft. Cytology identified round, large, plasmacytoid neoplastic cells. Histology revealed dense sheets of round neoplastic cells often surrounding or invading vascular walls (angiocentric/angiodestructive pattern). Neoplastic cells expressed CD3 (T cell phenotype) and FeLV p27 and gp70 antigens. A middle ear angiocentric angioinvasive T-cell lymphoma was diagnosed. Following radiation therapy, clinical conditions improved, but dysphagia recurred and the cat died suddenly. At necropsy examination, a soft, red mass filled the ventromedial compartment of the tympanic bulla. Extension to the base of the skull with right piriform lobe compression was recorded. Hepatic and splenic metastases were present. Conclusions: Diagnosis of primary middle ear tumors is often delayed since clinical signs mimic more common otological conditions. Multiple biopsy specimens and immunohistochemistry were pivotal for the diagnosis in this case. FeLV might have been involved in tympanic lymphoma development.
CANINE TESTICULAR TUMOURS: CORRELATION OF MITOTIC INDEX WITH Ki67 EXPRESSION G.F. Machado, J.M. Silva, G.D. Melo, A. Schweigert, J.E.S. Silva and F.G. Grano S~ao Paulo State University (UNESP), College of Veterinary Medicine, Laboratory of Applied Pathology, Arac¸atuba-SP, Brazil Introduction: Primary testicular tumours are common in older dogs. Histopathology and immunohistochemistry can provide relevant information about tumour cell proliferation, which can be useful to predict the prognosis. Therefore, this study aimed to evaluate and to compare Ki67 expression with the mitotic index in canine testicular tumours. Materials and Methods: Twenty-four seminomas, 12 Leydig cell tumours (LCTs) and five Sertoli cell tumours (SCTs) were included. The mitotic index was determined in 10 high-power microscopical fields (40 objective) in haematoxylin and eosin-stained tissue sections. Immunohistochemistry with the Ki67 antibody (clone MIB1, Dako M7240) was used to evaluate the Ki67-positive cells by computer-assisted image analysis. Results: Ki67 was expressed strongly by canine seminomas (2.9%), which was different to LCTs (0.2%) and SCTs (0.06%) (P !0.0001). The mitotic index was higher in seminomas (1.05%) than in LCTs (0.20%), but was not different from SCTs (0.60%) (P 5 0.0036). Comparing Ki67 immunoexpression with the mitotic index, a moderate positive correlation was observed in seminomas (r 5 0.483) and a perfect positive correlation in LCTs (r 5 1.000). Conclusions: The Ki67 index seems to be a more reliable tool to identify proliferating cells in canine testicular neoplasms than the visual identification of mitosis, leading to more accurate tumour grading.
ABERRANT EXPRESSION OF OCCLUDIN IN CANINE AND FELINE MAMMARY TUMOURS H. Warwick, L. Pritchard, K. Smith and I. McGonnell The Royal Veterinary College, London, United Kingdom Introduction: Occludin is a component of tight junctions, found in both mammary epithelia and myoepithelia. Reduction in occludin expression in epithelial cells is associated with malignancy in human mammary tumours. The aim of this study was to determine the expression of occludin in canine and feline mammary tumours. Materials and Methods: Eighteen canine and 19 feline mammary tumours were labelled using a specific anti-occludin antibody. Results: Benign tumours displayed normal occludin distribution, in the apical epithelium and the basal edge of the myoepithelium. However, nuclear occludin expression was also identified in these samples. Quantification of membrane occludin labelling showed a statistically significant reduction in occludin labelling in malignant tumours. Nuclear expression was also statistically significantly higher in malignant canine tumours, but not feline tumours. Examination of two mammary epithelial cell lines also showed nuclear occludin labelling that did not relate to the centromere on double labelling for gamma tubulin (centromere marker). Conclusions: Reduction in expression of both epithelial and myoepithelial occludin correlates with malignancy, which may point to the potential for occludin to act as a prognostic marker for mammary cancers. The appearance of nuclear occludin in these tissues is a new finding and further work in both tumour samples and breast cancer epithelial cells lines is required to elucidate its function in the disease process.
OVEREXPRESSION OF CYTOKERATIN 19, ALDOLASE A AND MANGANESE SUPEROXIDE DISMUTASE IN OVINE PULMONARY ADENOCARCINOMA A. Kycko and M. Reichert National Veterinary Research Institute in Pulawy, Poland Introduction: Ovine pulmonary adenocarcinoma (OPA) is a transmissible lung cancer of sheep caused by jaagsiekte sheep retrovirus. The present study aimed to compare the protein profiles of neoplastic and non-neoplastic lung tissues of sheep for the identification of potential protein biomarkers of OPA. Materials and Methods: Protein lysates obtained from frozen lung tissues of five OPA-affected sheep and four negative controls were analyzed by two-dimensional electrophoresis (2DE) and mass spectrometry. Expression of proteins identified in the spots presenting the highest overexpression in OPA proteomes was verified with immunohistochemistry (IHC). Results: As a result of 2DE analysis, 14 spots showed at least two-fold higher expression in each OPA sample compared with non-neoplastic tissue. In 11 out of 14 spots, more than one polypeptide was detected with mass spectrometry. The single proteins identified in three spots were cytokeratin 19, aldolase A and manganese superoxide dismutase. IHC revealed the expression of the three proteins in the cytoplasm of neoplastic cells in OPA, as well as in epithelial cells of bronchioles and type II pneumocytes in both non-neoplastic and OPA samples. Conclusions: The proteomic methods allowed for the identification of three polypeptides being part of the protein expression profile of OPA. Immunohistochemical examination of other identified proteins is still required to establish more markers of the disease.
ESVP/ECVP Proceedings 2013
111
PRIMARY ANGIOCENTRIC/ANGIOINVASIVE T-CELL LYMPHOMA OF THE TYMPANIC BULLA IN A FeLVPOSITIVE CAT S.F. Santagostino, C.M. Mortellaro, A. Forlani, G. Ghisleni and P. Roccabianca DIVET-Dipartimento di Scienze Veterinarie e Sanita Pubblica, University of Milano, Italy Introduction: Reports of primary middle ear lymphoma are rare in cats and their extension to internal organs is exceptional. A 5-yearold, neutered female, FeLV-positive domestic shorthair cat was referred for stertor, dyspnoea and head-tilt. CT scan revealed soft tissue opacity inside the right tympanic bulla with bone lysis and concurrent nasopharyngeal and intracranial invasion. Materials and Methods: Endoscopic-guided biopsy samples were collected for histology and immunohistochemistry. A full necropsy examination was performed. Results: Grossly, the lesion was poorly demarcated, white and soft. Cytology identified round, large, plasmacytoid neoplastic cells. Histology revealed dense sheets of round neoplastic cells often surrounding or invading vascular walls (angiocentric/angiodestructive pattern). Neoplastic cells expressed CD3 (T cell phenotype) and FeLV p27 and gp70 antigens. A middle ear angiocentric angioinvasive T-cell lymphoma was diagnosed. Following radiation therapy, clinical conditions improved, but dysphagia recurred and the cat died suddenly. At necropsy examination, a soft, red mass filled the ventromedial compartment of the tympanic bulla. Extension to the base of the skull with right piriform lobe compression was recorded. Hepatic and splenic metastases were present. Conclusions: Diagnosis of primary middle ear tumors is often delayed since clinical signs mimic more common otological conditions. Multiple biopsy specimens and immunohistochemistry were pivotal for the diagnosis in this case. FeLV might have been involved in tympanic lymphoma development.
CANINE TESTICULAR TUMOURS: CORRELATION OF MITOTIC INDEX WITH Ki67 EXPRESSION G.F. Machado, J.M. Silva, G.D. Melo, A. Schweigert, J.E.S. Silva and F.G. Grano S~ao Paulo State University (UNESP), College of Veterinary Medicine, Laboratory of Applied Pathology, Arac¸atuba-SP, Brazil Introduction: Primary testicular tumours are common in older dogs. Histopathology and immunohistochemistry can provide relevant information about tumour cell proliferation, which can be useful to predict the prognosis. Therefore, this study aimed to evaluate and to compare Ki67 expression with the mitotic index in canine testicular tumours. Materials and Methods: Twenty-four seminomas, 12 Leydig cell tumours (LCTs) and five Sertoli cell tumours (SCTs) were included. The mitotic index was determined in 10 high-power microscopical fields (40 objective) in haematoxylin and eosin-stained tissue sections. Immunohistochemistry with the Ki67 antibody (clone MIB1, Dako M7240) was used to evaluate the Ki67-positive cells by computer-assisted image analysis. Results: Ki67 was expressed strongly by canine seminomas (2.9%), which was different to LCTs (0.2%) and SCTs (0.06%) (P !0.0001). The mitotic index was higher in seminomas (1.05%) than in LCTs (0.20%), but was not different from SCTs (0.60%) (P 5 0.0036). Comparing Ki67 immunoexpression with the mitotic index, a moderate positive correlation was observed in seminomas (r 5 0.483) and a perfect positive correlation in LCTs (r 5 1.000). Conclusions: The Ki67 index seems to be a more reliable tool to identify proliferating cells in canine testicular neoplasms than the visual identification of mitosis, leading to more accurate tumour grading.
ABERRANT EXPRESSION OF OCCLUDIN IN CANINE AND FELINE MAMMARY TUMOURS H. Warwick, L. Pritchard, K. Smith and I. McGonnell The Royal Veterinary College, London, United Kingdom Introduction: Occludin is a component of tight junctions, found in both mammary epithelia and myoepithelia. Reduction in occludin expression in epithelial cells is associated with malignancy in human mammary tumours. The aim of this study was to determine the expression of occludin in canine and feline mammary tumours. Materials and Methods: Eighteen canine and 19 feline mammary tumours were labelled using a specific anti-occludin antibody. Results: Benign tumours displayed normal occludin distribution, in the apical epithelium and the basal edge of the myoepithelium. However, nuclear occludin expression was also identified in these samples. Quantification of membrane occludin labelling showed a statistically significant reduction in occludin labelling in malignant tumours. Nuclear expression was also statistically significantly higher in malignant canine tumours, but not feline tumours. Examination of two mammary epithelial cell lines also showed nuclear occludin labelling that did not relate to the centromere on double labelling for gamma tubulin (centromere marker). Conclusions: Reduction in expression of both epithelial and myoepithelial occludin correlates with malignancy, which may point to the potential for occludin to act as a prognostic marker for mammary cancers. The appearance of nuclear occludin in these tissues is a new finding and further work in both tumour samples and breast cancer epithelial cells lines is required to elucidate its function in the disease process.
OVEREXPRESSION OF CYTOKERATIN 19, ALDOLASE A AND MANGANESE SUPEROXIDE DISMUTASE IN OVINE PULMONARY ADENOCARCINOMA A. Kycko and M. Reichert National Veterinary Research Institute in Pulawy, Poland Introduction: Ovine pulmonary adenocarcinoma (OPA) is a transmissible lung cancer of sheep caused by jaagsiekte sheep retrovirus. The present study aimed to compare the protein profiles of neoplastic and non-neoplastic lung tissues of sheep for the identification of potential protein biomarkers of OPA. Materials and Methods: Protein lysates obtained from frozen lung tissues of five OPA-affected sheep and four negative controls were analyzed by two-dimensional electrophoresis (2DE) and mass spectrometry. Expression of proteins identified in the spots presenting the highest overexpression in OPA proteomes was verified with immunohistochemistry (IHC). Results: As a result of 2DE analysis, 14 spots showed at least two-fold higher expression in each OPA sample compared with non-neoplastic tissue. In 11 out of 14 spots, more than one polypeptide was detected with mass spectrometry. The single proteins identified in three spots were cytokeratin 19, aldolase A and manganese superoxide dismutase. IHC revealed the expression of the three proteins in the cytoplasm of neoplastic cells in OPA, as well as in epithelial cells of bronchioles and type II pneumocytes in both non-neoplastic and OPA samples. Conclusions: The proteomic methods allowed for the identification of three polypeptides being part of the protein expression profile of OPA. Immunohistochemical examination of other identified proteins is still required to establish more markers of the disease.