Malignant transformation of a giant congenital pigmented nevus (hamartoma) in a dog

A male Golden Retriever was born with a large area of abnormal skin and hair on the distal pelvic limb. A tumour arose from the proximal margin of this area at 5 years of age. Histopathological examination of the abnormal area of skin revealed an area in which follicles were surrounded by nodular accumulations of densely packed round to spindle-shaped cells with fine granular intracytoplasmic melanin. Similar cells were present within the subcutaneous fat, and clusters of densely pigmented melanocytes were scattered within the basal epidermis, follicular epithelium, and dermis. A diagnosis of giant congenital pigmented nevus (hamartoma) was made. The tumour from the proximal end of this area was composed of densely packed, moderately pleomorphic, poorly differentiated and pigmented, spindle-shaped to epithelioid melanocytes, and was diagnosed as malignant melanoma. Metastasis of the malignant melanoma to the stifle and inguinal regions occurred within 6 months. To date, the authors are unaware of prior reports of a canine giant congenital pigmented nevus (hamartoma) with transformation to a malignant melanoma.     

Osteoclast-like giant cell tumour arising from the kidney in a dog

In this study, a case of osteoclast-like giant cell tumour arising from the kidney is reported in an eight-year-old female Anatolian Shepherd dog. Macroscopically, the tumorous mass covered the hilus of the left kidney. It was 26 x 22 x 12 cm in size and 3700 g in weight. Metastatic tumorous nodules, 0.5-2.0 cm in diameter, were found on the abdominal side of the diaphragm and in the lungs. Microscopically, numerous large osteoclast-like multinucleated giant cells and spindle-spheroidal-shaped cells were seen. Osteoblastic differentiation and osteoid matrix were noted in a few areas at the periphery of the tumour, near the connective tissue septa. The stroma of the tumour tissue was vascular, oedematous and loose. By immunoperoxidase staining, tumour cells showed immunoreactivity for vimentin but not for keratin and desmin, indicating that the tumour had mesenchymal origin. This is the first report in the literature on a malignant osteoclast-like giant cell tumour arising from a visceral organ in animals.     

Orbitotomy for retrobulbar malignant fibrous histiocytoma in a dog

A retrobulbar malignant fibrous histiocytoma was diagnosed in a 12-year-old castrated male Keeshond dog. The mass was excised with a lateral orbitotomy and zygomatic arch resection. Vision was preserved in the affected eye, and no recurrence was noted up to 10 months postoperatively. Malignant fibrous histiocytoma originates from primitive mesenchymal stem cells. The malignant fibrous histiocytoma seen in our patient was most consistent with the storiform-pleomorphic variant, given the storiform arrangement of spindle cells, the presence of histiocytoid cells, and a mixed inflammatory infiltrate, without giant cells. The metastatic potential of malignant fibrous histiocytoma in general, and the storiform variant in particular, is unknown. Seventeen months later the dog was presented to the referring veterinarian with anorexia, diarrhea, weight loss and bilateral purulent nasal exudates. The dog was euthanized without necropsy.     

Chondroitin sulfate proteoglycan-4: a biomarker and a potential immunotherapeutic target for canine malignant melanoma

Chondroitin sulfate proteoglycan-4 (CSPG4), also known as high molecular weight-melanoma associated antigen (HMW-MAA), is a membrane-bound chondroitin sulfate proteoglycan highly expressed by human melanoma cells. This phylogenetically conserved tumour antigen plays an important biological role in human melanoma, where it is used as a marker to diagnose forms with unusual characteristics, such as desmoplastic melanoma, and to detect melanoma cells in lymph nodes and peripheral blood, and as a target for immunotherapy because of its restricted distribution in normal tissues. To identify suitable targets to develop novel approaches of treating canine melanoma, CSPG4 was studies to see whether it is expressed in canine malignant melanomas. Immunohistochemical staining of 65 canine malignant melanomas with an anti-human CSPG4-specific antibody detected CSPG4 in 37 cases (56.9%). Positive staining was more frequent, albeit not significantly, in amelanotic compared to melanotic tumours and was statistically associated with tumours having both melanin and the epithelioid histotype. The frequency of CSPG4 expression was similar to that of other melanoma antigens used as diagnostic markers for canine malignant melanoma, such as Melan A and the protein recognized by the PNL2 monoclonal antibody. The results suggest that CSPG4 constitutes a new potential immunohistochemical marker of canine malignant melanoma and may represent an immunotherapeutic target as in humans.     

Induction of dendritic cell-mediated immune responses against canine malignant melanoma cells

To establish the basis for the use of dendritic cells (DC) in the treatment of canine melanoma, dogs were vaccinated using autologous DC pulsed with canine melanoma CMM2 cell lysate in the presence of keyhole limpet haemocyanin (KLH) in vitro (CMM2-KLH-DC), and the induction of immune responses against CMM2 cells in vivo was examined using the delayed-type hypersensitivity (DTH) skin test. The DTH responses against CMM2 cells and KLH were observed in dogs vaccinated with CMM2-KLH-DC, while the responses against KLH but not CMM2 cells were detected with DC pulsed with KLH alone (KLH-DC). Recruitment of CD8 and CD4 T cells was detected in the positively responding sites, suggested that vaccination with CMM2-KLH-DC efficiently elicits T cell-mediated immunity against CMM-2 cells in vivo. These findings demonstrate the potential utility of DC-based tumour vaccination in the treatment of canine malignant melanoma.     

Malignant fibrous histiocytoma (giant cell type) associated with a malignant mixed tumor in the salivary gland of a dog

A 12-year-old male Boxer dog presented with a 5 x 5 x 7-cm partially encapsulated mass in the right mandibular salivary gland. Histologically, the mass was composed of neoplastic epithelial and mesenchymal cells. The mesenchymal component consisted of two cell populations arranged in different patterns: coalescing nodules of neoplastic mononuclear cells with rare osteoid and numerous osteoclastlike giant cells; and sheets of neoplastic spindle cells intermingled with neoplastic epithelial cells and containing osteoid and well-formed bone trabeculae lined by osteoblasts and few osteoclastlike giant cells. On the basis of these histological features, two malignant salivary tumors were diagnosed: a malignant fibrous histiocytoma (giant cell type) and a malignant mixed tumor. Immunohistochemical studies demonstrated keratin 5 and 8 expression by the neoplastic epithelial cells, indicating a probable salivary ductal origin, and vimentin expression by all mesenchymal elements, suggesting a fibroblastic line of differentiation.     

Carcinosarcoma mimicking a feline injection‐site sarcoma in a cat

A 15‐year‐old spayed female domestic short‐haired cat with cutaneous/subcutaneous well‐circumscribed, alopecic mass approximately 25 × 30 mm in diameter, localized to the left shoulder region was brought to the veterinary surgery department. Despite the suggestive location and macroscopic appearance, feline injection‐site sarcoma was not suspected based on the cytologic examination of fine‐needle aspirates. The tumor was surgically resected, and tissue sections were evaluated microscopically. The tumor was found to be nonencapsulated with a distinct border between the neoplastic parenchyma and surrounding connective tissue. The neoplastic tissue consisted of 2 cell populations: elongated to spindle‐shaped cells arranged in bands and cords and malignant epithelial‐like cells. Both populations showed microscopic features of malignancy. Multinucleate giant cells with irregular cytoplasm were scattered among the neoplastic cells. The spindle‐shaped cells strongly expressed vimentin but did not express α‐smooth muscle actin (α‐SMA) or cytokeratin. Desmin was strongly expressed in about 0‐5% of cells. Epithelial‐like cells expressed cytokeratin, but not vimentin, desmin, or α‐SMA. Multinucleate giant cells expressed vimentin, but did not α‐SMA, desmin, or cytokeratin. Based on microscopic observations and IHC results, the final diagnosis was carcinosarcoma with histologic features compatible with feline injection‐site sarcoma, but without the clinical aggressiveness of this tumor.     

Juvenile malignant mesothelioma in a dog

An 11-month-old male mixed breed dog was euthanized due to two months history of vomiting and anorexia. At necropsy, numerous, multifocal or coalescing, firm, protruding nodules, 5 to 40 mm in diameter were scattered throughout the mesentery and omentum. Histologically and immunohistochemically, the nodules were diagnosed as malignant mesothelioma. Metastasis to the regional mesenteric, mediastinal and tracheobronchial lymph nodes were observed.     

A salivary malignant myoepithelioma in a dog

Salivary tumours are uncommon in domestic animals and there are no known previous confirmed reports of salivary tumours of myoepithelial origin in dogs. A 12-year-old female mixed breed dog was presented with a lobulated mass, composed of white-yellowish tissues, extending from soft palate to epiglottis. Histological examination revealed a neoplastic lesion consisting of a dense population of cells showing moderate pleomorphism, with pale cytoplasm and large oval nuclei, arranged in solid lobules. Mitotic activity was very high. Tumoral cells were negative for both periodic acid-Schiff reaction and Alcian blue stain and displayed strong immunohistochemical reactivity for pan-cytokeratin, muscle specific actin and myosin and focal positivity for cytokeratin 14. On the basis of the morphological, histochemical and immunohistochemical findings a diagnosis of malignant tumour of myoepithelial origin (malignant myoepithelioma) was made.     

Serum neopterin levels in female dogs with malignant mammary tumours

In this study, we have determined serum neopterin levels in female dogs with primary malignant mammary tumours. The study involved 50 female dogs which had a malignant mammary tumours removed surgically (32 animals with carcinoma, 12 animals with sarcoma and 6 animals with carcinosarcoma) and 10 clinically healthy female dogs. Serum neopterin levels were determined using a commercial ELISA kit. The mean neopterin levels were lower in the malignant tumour groups than in healthy animals but differences were statistically significant only in carcinoma and sarcoma groups. The decrease of neopterin levels in animals with malignant mammary tumours may suggest their decreased cellular immunity. Moreover, it might indicate that decreased activity of cellular mechanisms of the anti‐neoplastic response is one of the factors associated with the development and course of malignant mammary tumours in female dogs; however, further studies are necessary.     

Canine oral malignant melanoma: Prognostic utility of an alternative staging system

Reports of prognosis based on clinical staging of canine oral malignant melanoma consistently differ. To determine the prognostic utility of the World Health Organization (WHO) tumour, node, metastasis (TNM) system and a proposed alternative staging system, a retrospective study of 41 dogs with naturally occurring oral malignant melanoma was conducted. All the tumours were clinically staged, removed surgically and the tissues histologically reviewed. Treatment responses were correlated with the clinical and histological features reported to have prognostic utility. Dogs presenting with a tumour smaller than 8 cm3, located on the rostral mandible or caudal maxilla, and, or, having a tumour mitotic index of 3 or less had a significantly longer remission length and survival time than did other dogs, regardless of the treatment selected. Treatment by radical surgical excision (eg, hemimandibulectomy) resulted in longer remission lengths and survival times than any other type of treatment regardless of whether or not surgical margins were determined to be free of residual tumour. The WHO system failed to identify a prognostic difference between dogs of a differing clinical stage. An alternative system derived from significant features mentioned above did identify a prognostic difference and is recommended for further evaluation regarding its utility in the pre-treatment evaluation and clinical staging of other dogs with oral -malignant melanoma.     

Effects of low‐dose cyclophosphamide with piroxicam on tumour neovascularization in a canine oral malignant melanoma‐xenografted mouse model

Low‐dose cyclophosphamide (CyLD) has shown promise in the treatment of several cancers; however, the effect of CyLD on canine oral malignant melanoma has never been explored. In this study, we investigated the effects of CyLD with or without piroxicam (Px) on tumour neovascularization and vascular normalization in a canine oral malignant melanoma‐xenografted mice model. After treatment with CyLD, Px or a combination of both (CyPx), the growth of the tumour in the treatment groups was significantly suppressed compared to the control group at 30 days of treatment. Proliferation index was also significantly reduced by all treatments, only CyPx significantly lowered microvessel density and vascular endothelial growth factor (VEGF) levels. Additionally, CyLD significantly reduced the proportion of normal vessels and caused an imbalance between VEGF and thrombospondin‐1. These results suggested that CyPx has potent anti‐angiogenic effects in terms of both the number and quality of blood vessels in xenografted canine oral malignant melanoma.     

Conjunctival malignant melanoma in a horse

A case of malignant melanoma originating from the conjunctiva of a horse is reported. The tumor exhibited locally aggressive behavior as evidenced clinically by recurrence following two treatment episodes including surgical excision on each occasion and one application of cryotherapy. The orbit was subsequently exenterated and histologically malignant conjunctival melanoma was confirmed. Histopathologic features included variable pigmentation with amelanotic sites demonstrating marked cellular and nuclear pleomorphism with high numbers of mitotic figures. Cords of neoplastic cells invaded the sclera and cornea. Following exenteration, the horse exhibited no recurrence of the tumor for five years before being lost to follow-up. To our knowledge, this is the first report of primary malignant conjunctival melanoma in a horse.     

An ultrastructural study of malignant mesotheliomas in two cows

Malignant mesotheliomas of two Holstein cows were examined by light and electron microscopy. These tumours were derived from peritoneal surfaces and were of biphasic type composed of mesothelial lining cells and submesothelial mesenchymal cells. Ultrastructurally, the neoplastic mesothelial cells were characterized by intermediate filaments, microvilli, desmosomes, tight junctions and basal laminae. The neoplastic submesothelial cells were closely associated with collagen filaments and some cells had several resemblance to the neoplastic mesothelial cells. The ultrastructural features of the mesotheliomas are discussed in comparison with those of adenocarcinomas and fibroblastic neoplasms. In one case of malignant granulosa cell tumour involved the left ovary and its morphology was apparently different from that of the mesothelioma. Multiple primary tumours are uncommon in cattle.     

Metastasizing fibrosarcoma in a Wistar rat--case report

A 19-month-old male control Wistar rat from a 30-month inhalation study showed a subcutaneous greyish-white mass extending from the throat to the thoracic cavity. The rat had been euthanized because of its poor general condition. Histologically, the mass was diagnosed as a fibrosarcoma infiltrating the masseter muscle with metastases in the lungs, liver and heart. The primary tumour was characterized by fusiform spindle cells producing various amounts of interlacing bundles of collagen. The cells formed a characteristic herringbone pattern and mitotic figures were frequent. The histological parameters of the metastases were practically identical to those seen in the primary tumour. The diagnosis was confirmed by trichrome staining and positive immunostaining for vimentin and was differentiated from leiomyosarcomas by its negative immunostaining for desmin, from schwannomas by its negative immunostaining for S-100 and from malignant fibrous histiocytomas by the absence of giant cells. The incidence of fibrosarcomas in Wistar rats is very low (up to 3%) and metastasis is rarely observed.     

Osteoclastic-like giant cell tumour in a cat

An 11-year-old neutered male domestic shorthair cat was presented with an epulis. A hemispherical mass, 8mm in maximum diameter, without a peduncle and bright reddish in colour, was observed on the gingiva of the left mandible. Radiography failed to show any infiltrating osteolysis. The epulis was surgically removed via gingival incision around the margin to the depth of connective tissue layer. Histopathological examination indicated that the epulis contained a large number of multinucleated giant cells (MGCs) intermixed with mononuclear mesenchymal cells in a loose fibrovascular stroma. Mitotic cells were found, mainly in the centre of the mass. MGCs were stained positive by the tartrase resistant acid phosphatase (TRAP) staining, indicating osteoclasts activity. Immunohistochemical staining for proliferating-cell nuclear antigen (PCNA) was observed within the majority of mononucleated cells, whereas multinucleated cells did not stain. An osteoclast-like giant cell tumour was concluded in this case. The origin of epulis is likely from the periosteal tissue. The cat recovered uneventfully and no recurrence has been noted for 3 years thereafter.     

Paravertebral malignant peripheral nerve sheath tumour (MPNST) in a horse

A 10‐year‐old Lipizzaner gelding was presented with intermittent ataxia and hindlimb weakness. Ultrasonographic examination identified a mass cranial to the tuber sacrale. A provisional diagnosis of a soft tissue sarcoma was made based on a biopsy specimen. Owing to the extensive nature of the tumour and the associated poor prognosis, the horse was subjected to euthanasia on humane grounds. A post mortem examination revealed a locally infiltrative soft mass within the left lumbosacral epaxial musculature. Histologically, an infiltrative neoplasm predominantly composed of pleomorphic spindle or stellate‐shaped cells was identified. Neoplastic cells exhibited strong S‐100 protein and GFAP expression and variable vimentin, NSE, NGFR and myoglobin expression. They were uniformly negative for pan‐cytokeratin, melan A, laminin and desmin. The ultrastructural examination revealed pleomorphic cells with long cytoplasmic processes and an absence of melanosomes. Based on these results, a diagnosis of malignant peripheral nerve sheath tumour was made. This report contributes further information to assist in the diagnosis of these poorly defined neoplasms in animals, especially when they occur in uncommon locations.     

Pathology of canine oral malignant melanoma with cartilage and/or osteoid formation

Of 197 cases of canine oral malignant melanoma, 29 cases with myxoid, cartilage, and osteoid formation were studied pathologically and immunohistochemically. Tumor tissues were classified into spindle cell type (13 cases), epithelioid cell type (1 case), and mixed type (15 cases). Myxoid matrixes (29 tumors) were formed mainly in the tissues of spindle cell type and were positive for Alcian blue (pH 2.5). Cartilaginous matrixes (12 tumors) were formed in the myxoid tumor tissues. The morphology of atrophied neoplastic cells, which were embedded in the cartilage cavities, significantly differed from that of spindle cells proliferating in surroundings. There were reticular areas in the process of transitioning from myxoid to cartilaginous matrixes. Osteoid matrixes were not continuous with myxoid or cartilaginous matrixes, and arose as eosinophilic trabeculae in the dense collagenous connective tissues. A calcified bone trabecula was present among the osteoid trabeculae in a case. Melanin-producing melanocytes were proliferating in the collagenous matrixes, while amelanotic cells were in the osteoid matrixes. Immunohistochemistry demonstrated proliferating neoplastic cells as melanocytes. All cells in/out of these three matrixes were positive for Melan-A, S-100 protein, NSE, and vimentin. From these results, it is suggested that cartilage and osteoid matrixes are produced by dedifferentiated melanocytes.     

Comparative study of anti-oncogenic microRNA-145 in canine and human malignant melanoma

MicroRNA-145 (miRNA-145; miR-145) is aberrantly expressed in most of human cancers and plays a significant role in carcinogenesis and cancer progression. In the current study, we focused on how miR-145 plays a role in canine and human malignant melanomas. MiR-145 was significantly downregulated in canine malignant melanoma tissues and canine melanoma cell lines, as well as human melanoma cell lines tested. The ectopic expression of miR-145 showed a significant growth inhibition in both canine and human melanoma cells tested, and the effect was achieved partly by suppressing c-MYC in canine melanoma LMeC and in human melanoma A2058 and Mewo cells. At the same time, a suppressive tendency on cell migration in canine melanoma KMeC cells and significant suppression of cell migration in human melanoma A2058 cells by suppressing FASCIN1 were also found. These findings suggest that miR-145 acts as a tumor suppressor in both canine and human malignant melanomas.