The Molecular Basis of Canine Melanoma: Pathogenesis and Trends in Diagnosis and Therapy

Melanoma is a common neoplastic disease of dogs with variable presentation and biological behavior. Canine malignant melanoma is a rapidly metastatic disease that generally is incurable. The loss of function of cellular safeguards built into the genetic program and of immune surveillance systems that cooperate to prevent tumor formation and progression appear to be important underlying causes of canine malignant melanoma. In effect, many existing cancer treatments restore the function of 1 or the other of these mechanisms. For example, chemotherapy and radiotherapy often kill tumor cells by initiating a genetic suicide mechanism (apoptosis), and immunotherapy initiates or enhances a response by the body's immune cells to identify and destroy cancer cells by mechanisms that rely on direct cytotoxicity or apoptotic cell death. Nevertheless, standard therapeutic approaches have not proved effective in treatment of canine malignant melanoma, with only marginal improvement in the outcome of dogs with this disease. The advantages of an improved understanding of the molecular basis of canine cancer are underscored by recent promising advances in diagnosis and in immunologic and genetic therapies that may help reduce the mortality of dogs affected with malignant melanoma.     

MicroRNAs as tumour suppressors in canine and human melanoma cells and as a prognostic factor in canine melanomas

Malignant melanoma (MM) is one of the most aggressive cancers in dogs and in humans. However, the molecular mechanisms of its development and progression remain unclear. Presently, we examined the expression profile of microRNAs (miRs) in canine oral MM tissues and paired normal oral mucosa tissues by using the microRNA‐microarray assay and quantitative RT‐PCR. Importantly, a decreased expression of miR‐203 was significantly associated with a shorter survival time. Also, miR‐203 and ‐205 were markedly down‐regulated in canine and human MM cell lines tested. Furthermore, the ectopic expression of miR‐205 had a significant inhibitory effect on the cell growth of canine and human melanoma cells tested by targeting erbb3. Our data suggest that miR‐203 is a new prognostic factor in canine oral MMs and that miR‐205 functions as a tumour suppressor by targeting erbb3 in both canine and human MM cells.     

Malignant melanoma in a zebra finch (Taeniopygia guttata): cytologic, histologic, and ultrastructural characteristics

An approximately 3-year-old adult male zebra finch (Taeniopygia guttata) was diagnosed with malignant melanoma. The large darkly pigmented tumor was located in the coelom, extended from the apex of the heart to the cloaca, and was adherent to the intestines and the ventriculus. Dark small masses (likely metastases) were observed in the lungs. Cytologically, the neoplasm consisted mainly of round to oval cells with brown or pale blue to blue-brown pigment. Lesser numbers of cells were stellate to dendritic with abundant amounts of brown pigment granules or were markedly pleomorphic with variable amounts of pigment. Histologically, the tumor consisted of dense sheets and aggregates of infiltrative melanocytes that were negative for S-100 and Melan A. A few cells were consistent with "signet-ring" melanocytes. Melanocytes examined by electron microscopy contained typical structures, mainly premelanosomes and melanosomes, of this cell type. However, melanocytes with marked pleomorphism also contained intracytoplasmic aggregates of filaments, consistent with previously reported ultrastructural findings in signet-ring or rhabdoid melanoma of nonavian species.     

The oncolytic effects of reovirus in canine solid tumor cell lines

Oncolytic virotherapy is a new strategy for cancer treatment for humans and dogs. Reovirus has been proven to be a potent oncolytic virus in human medicine. Our laboratory has previously reported that canine mast cell tumor and canine lymphoma were susceptible to reovirus. In this study, canine solid tumor cell lines (mammary gland tumor, osteosarcoma and malignant melanoma) were tested to determine their susceptibility towards reovirus. We demonstrated that reovirus induces more than 50% cell death in three canine mammary gland tumors and one canine malignant melanoma cell line. The reovirus-induced cell death occurred via the activation of caspase 3. Ras activation has been shown to be one of the important mechanisms of reovirus-susceptibility in human cancers. However, Ras activation was not related to the reovirus-susceptibility in canine solid tumor cell lines, which was similar to reports in canine mast cell tumor and canine lymphoma. The results of this study highly suggest that canine mammary gland tumor and canine malignant melanoma are also potential candidates for reovirus therapy in veterinary oncology.     

Canine digital tumors: a veterinary cooperative oncology group retrospective study of 64 dogs

We compared clinical characteristics and outcomes for dogs with various digital tumors. Medical records and histology specimens of affected dogs from 9 veterinary institutions were reviewed. Risk factors examined included age, weight, sex, tumor site (hindlimb or forelimb), local tumor (T) stage, metastases, tumor type, and treatment modality. The Kaplan-Meier product limit method was used to determine the effect of postulated risk factors on local disease-free interval (LDFI), metastasis-free interval (MFI), and survival time (ST). Outcomes were thought to differ significantly between groups when P < or = .003. Sixty-four dogs were included. Squamous cell carcinoma (SCC) accounted for 33 (51.6%) of the tumors. Three dogs presented with or developed multiple digital SCC. Other diagnoses included malignant melanoma (MM) (n = 10; 15.6%), osteosarcoma (OSA) (n = 4; 6.3%), hemangiopericytoma (n = 3; 4.7%), benign soft tissue tumors (n = 5; 7.8%), and malignant soft tissue tumors (n = 9; 14%). Fourteen dogs with malignancies had black hair coats, including 5 of the 10 dogs with MM. Surgery was the most common treatment and, regardless of the procedure, had a positive impact on survival. None of the patient variables assessed, including age, sex, tumor type, site, and stage, had a significant impact on ST. Both LDFI and MFI were negatively affected by higher T stage, but not by type of malignancy. Although metastasis at diagnosis correlated with a shorter LDFI, it did not have a significant impact on ST. On the basis of these findings, early surgical intervention is advised for the treatment of dogs with digital tumors, regardless of tumor type or the presence of metastatic disease.     

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.     

Tumor stem cells and implications for the immunotherapy of cancer

Passive, active and adoptive immunotherapy of human cancers and leukemias raise a renewed interest strengthened by the availability of purified biological substances such as IL2, interferon, thymic hormones and by some recent favourable therapeutic results which have demonstrated the possibility of obtaining objective tumor regressions by immunotherapy. Analysis of results of chemotherapy shows that only 8 human cancers and malignant hemopathies are curable at an advanced diffuse stage of disease. These cancers are characterized by the rarity or absence of late metastases (more than 3-4 years after initial diagnosis). Cure may be considered as complete with a high probability if disease free status is maintained for 3 years. This finding suggests the absence of tumor stem cells capable to produce late metastases. Other cancers are not chemocurable at an advanced systemic stage. In most of them late metastases (greater than 4 years after diagnosis) are observed. A model of organization of malignant tumors based on the distinction between primitive tumor stem cells which are rarely or exceptionally in cycle and protected by a specific microenvironment and committed tumor stem cells is proposed. According to this model, only cancers and/or metastases and malignant hemopathies containing but committed tumor stem cells would be chemocurable. Analysis of a trial of adjuvant therapy of breast cancer with poly A: poly U shows the possibility of immunotherapy to prevent the development of late metastases, independently of hormonal status in contrast with standard adjuvant chemotherapy which is only active on early micrometastases in premenopausal women.(ABSTRACT TRUNCATED AT 250 WORDS)     

Malignant melanoma in a Eurasian otter (Lutra lutra).

An 11-yr-old female Eurasian otter (Lutra lutra) presented with multiple cutaneous nodules identified histologically as malignant melanomas of spindle cell and epithelioid cell type. Metastases were detected in lymph nodes and liver, and the tumor, which was derived from melanocytes, showed aggressive biological behavior. Only occasional reports exist of neoplastic disease in otters.     

Malignant melanoma in pleural effusion in a 14-year-old cat

CASE DETAILS: A 14-year-old female cat presented with signs of respiratory distress. Pleural fluid was found on radiographic assessment. Cytologic evaluation of the fluid revealed malignant melanocytosis. The cat had a previous history of a recurrent malignant melanoma near the base of the right ear. Due to declining clinical condition, the cat was euthanized. CLINICAL SIGNIFICANCE: Cutaneous malignant melanomas (or melanosarcomas) are uncommon neoplasms in cats, and knowledge is limited. As far as the authors are aware, there are no previous reports in the veterinary literature of malignant melanocytes being identified in pleural effusion in cats, as they have in dogs. This report suggests that, despite conflicting information in the literature regarding the clinical behavior of cutaneous melanomas in cats, these tumors are capable of recurrence and metastasis. Aggressive treatment may be necessary even, as in this case, if the tumor is well differentiated on histopathology.     

A comparative review of melanocytic neoplasms

Melanoma is a devastating disease frequently encountered within both veterinary and human medicine. Molecular changes linked with neoplastic transformation of melanocytes include mutations in genes that encode proteins intrinsic to the regulatory pathways of two tumor suppressor proteins (retinoblastoma protein and p53), proto-oncogene mutation to oncogenes, altered expression of epithelial cadherin and CD44 adhesion molecules, and upregulation of angiogenic factors and other growth factors. Histologic evaluation of the primary mass is the most common means of diagnosis, with cytology used more frequently to document metastasis. Melanoma's highly variable histologic and cytologic patterns can make diagnosis by either method problematic. Adherent epithelioid morphology, including signet ring forms, and nonadherent round and spindle forms are recognized, with pigmentation an inconsistent finding. The site of the tumor, the thickness of the primary tumor or depth of invasion, and the number of mitotic figures per high-power field or per millimeter are used histologically to predict biologic behavior, whereas site and degree of pleomorphism are typically used for cytologic preparations. Diagnosis of amelanotic melanoma can be aided by ancillary diagnostic techniques. Tumor cells are usually positive for vimentin, S100, neuron-specific enolase, and Melan-A, and negative for cytokeratin. Melan-A as a positive marker is not as sensitive as the others are but is likely more specific. Monoclonal antibodies to human melanosome-specific antigens 1 and 5 cross-react with canine antigens for a combined sensitivity rate of 83%. Mouse monoclonal antibody IBF9 specifically recognizes canine melanoma antigen and also has good sensitivity. Serologic markers, including cytokines, cell adhesion molecules, and melanoma-inhibitory activity, are being investigated as potential sentinels of melanoma. Currently, there is no single diagnostic technique capable of differentiating benign from malignant melanocytic neoplasms or predicting survival time.     

Hemangiosarcoma in dogs and cats.

Hemangiosarcoma (HSA, including angiosarcoma and malignant hemangioendothelioma) is a highly malignant tumor derived from the endothelial cell line and is characterized by early and aggressive metastasis. HAS is a common tumor type in dogs, but is rare in other species. Treatment options include surgery, chemotherapy, and possibly radiation, but survival times are invariably short (usually < 1 year), except for patients with superficial dermal HSAs. Further options to treat this tumor type are currently being investigated.     

Molecular carcinogenesis of canine mammary tumors: news from an old disease

Studies focusing on the molecular basis of canine mammary tumors (CMT) have long been hampered by limited numbers of molecular tools specific to the canine species. The lack of molecular information for CMT has impeded the identification of clinically relevant tumor markers beyond histopathology and the introduction of new therapeutic concepts. Additionally, the potential use for the dog as a model for human breast cancer is debatable until questions are answered regarding cellular origin, mechanisms, and cellular pathways. During the past years, increasing numbers of canine molecular tools have been developed on the genomic, RNA, and protein levels, and an increasing number of studies have shed light on specific aspects of canine carcinogenesis, particularly of the mammary gland. This review summarizes current knowledge on the molecular carcinogenesis of CMT, including the role of specific oncogenes, tumor suppressors, regulators of apoptosis and DNA repair, proliferation indices, adhesion molecules, circulating tumor cells, and mediators of angiogenesis in CMT progression and clinical behavior. Whereas the data available are far from complete, knowledge of molecular pathways has a significant potential to complement and refine the current diagnostic and therapeutic approach to this tumor type. Furthermore, current data show that significant similarities and differences exist between canine and human mammary tumors at the molecular level. Clearly, this is only the beginning of an understanding of the molecular mechanisms of CMT and their application in clinical patient management.     

The biology of melanocytes

In veterinary medicine, our understanding of the biology and regulation of melanocytic function is mostly based on information realized from human and murine studies. Improved understanding of the biology of melanocytes is needed to develop more effective treatment regimens for malignant melanoma and other melanocytic disorders. In vertebrates, melanocytes are well known for their role in skin pigmentation, hair and feather coloration, and for their ability to produce and distribute melanin to surrounding keratinocytes. Enzymes involved in melanin synthesis are present exclusively in melanosomes. The type of melanin synthesized by melanocytes in mammals is regulated at a genetic, biochemical and environmental level. These regulatory factors affect not only the phenotypic appearance, but also the photoprotective properties of melanin. This review addresses the biology of melanocytes, melanin synthesis and the photoprotective properties of melanin.     

Genetic and environmental aspects of the immune responseAspects de la résponse immunitaire liés à la génétique l'environmentGenetische und umweltbedingte aspekte der immunreaktion

The main characteristic of the immune system is the potency to detect and resist invasions of foreign substances such as viruses, bacteria, parasites and transformed malignant cells. This ability is characterized and determined mostly by macrophage activity and T and B lymphocytes.These three major components of the immune response are independently genetically regulated. This makes selection on the basis of the immune response difficult.Environmental factors can induce stress situations which result in high blood corticosteroid levels. Corticosteroids can be immunosuppressive. In modern animal husbandry practice such factors exist and can influence disease resistance resulting in multifactorial diseases.     

动物黑色素细胞及其相关性状形成机制的研究进展

黑色素细胞是合成黑色素的细胞，由其前体细胞黑色素母细胞分化而来，胚胎期黑色素母细胞的形成有两次，一次是具有黑色素细胞特性的神经嵴细胞沿背外侧迁移而形成，另一次是由沿腹侧迁移的神经嵴细胞亚细胞雪旺氏细胞前体（Schwann cell precursors，SCPs）受到某种信号诱导后形成，这是皮肤中黑色素细胞形成的主要方式。黑色素细胞的形成与动物毛发颜色及其他重要经济性状密切相关，其形成的增多或减少，异位形成或异位侵袭均可能导致相应的疾病，但脊椎动物中具有典型黑色素细胞异位形成特征的乌骨鸡却繁衍至今，而且是重要的食用和药用经济动物。探索黑色素细胞的形成及其对相关性状的影响不但能为动物经济性状育种提供相关理论基础，也可为相关疾病的治疗提供理论依据。作者从黑色素细胞的来源及其形成过程中参与的信号因子来探讨其形成机制，并对黑色素细胞相关性状的形成机制尤其是乌质性状进行综述。     

溶瘤病毒在犬肿瘤治疗中的研究进展

溶瘤病毒疗法是利用病毒治疗恶性肿瘤的一种较有前景的新型生物疗法。溶瘤病毒不仅能直接特异性感染和溶解肿瘤细胞，也能刺激肿瘤部位的免疫反应来间接地溶解肿瘤组织细胞，且对正常组织细胞无杀伤或仅有较弱的杀伤作用，从而能达到理想的抗肿瘤效果。文章综述了溶瘤病毒的发展史及溶瘤病毒介导肿瘤消融术的机制，分析麻疹病毒、犬瘟热病毒、新城疫病毒、仙台病毒等8种病毒作为抗肿瘤药物在体外试验、异种移植肿瘤动物模型试验、肿瘤患犬治疗试验等不同阶段对各种犬肿瘤细胞的杀伤效果，提示溶瘤病毒疗法可能是一种安全可靠的恶性肿瘤治疗新方法，但真正将溶瘤病毒疗法应用在犬肿瘤临床治疗中时病毒肿瘤特异性、病毒使用剂量等问题仍需解决，本综述可为溶瘤病毒在犬肿瘤临床治疗中的安全使用提供新的思路和技术途径。     

Pathogenicity of Mycoplasma meleagridis for chicken cells

Mycoplasma meleagridis (MM) is a known pathogen for turkeys only. In this study, MM was used to inoculate chicken embryos and tumor cells to assess its pathogenic potential for chickens. In chicken embryos, it caused abnormal-shaped toes and severely denuded tracheae. In chicken tumor cells, MM reduced the cellular capacity to release a chemoattractant that causes the migration of heterophils. MM also caused death and/or a reduced growth rate in chicken HD-11 cells, a macrophage-monocyte-derived cell line. Thus, the data show that MM is a potential pathogen for chicken embryos and chickens cells. Further exploration to determine the pathogenicity in chickens may be warranted.     

Quercetin induces structural chromosomal aberrations and uncommon rearrangements in bovine cells transformed by the E7 protein of bovine papillomavirus type 4

Bovine papillomavirus type 4 (BPV‐4) and bracken fern are cofactors in the carcinogenesis of the upper gastrointestinal (GI) tract of cattle. An experimental in vitro model system has been developed to analyse the co‐operation between the viral transforming protein E7, the cellular ras oncogene and quercetin, one of the mutagens of bracken fern, during neoplastic progression of primary bovine cells. We now report cytogenetic studies of these cells at different stages of malignant transformation: parental primary non‐transformed PalF cells; E7R cells transformed by BPV‐4 E7 and activated ras but not tumorigenic, and tumorigenic E7Q cells derived from E7R cells after treatment with quercetin. All cell lines presented increased numbers of aneuploid cells. The rate of structural chromosomal aberrations observed was increased in transformed cells. In addition, E7Q cells showed chromosomes with peculiar rearrangements, which resulted in metacentric and submetacentric marker chromosomes, with an increase in the mean chromosome arm number. These markers were the products of possible centric fusions. These aberrations and rearrangements were distributed throughout the karyotype, no specific chromosome was involved and the heterochromatic centromeric regions appeared to be preserved.