Primary malignant melanoma in the oesophagus of a foal

A 2‐month‐old filly was evaluated for severe colic. Ultrasound, abdominocentesis and physical examination findings prompted an abdominal exploratory surgery. Perforation of the stomach was discovered during the surgery. The filly was humanely subjected to euthanasia under anaesthesia and post mortem examination was performed. In addition to gastric and duodenal ulceration, a thickened, black area of the proximal oesophagus was discovered. Histopathology of the lesion revealed primary malignant melanoma. Although rare, primary melanoma can occur in noncutaneous locations.     

Surgical treatment of epibulbar melanocytomas by complete excision and homologous corneoscleral grafting in dogs: 11 cases

Objective The aim of the study was to evaluate the efficacy, outcome, and complications following penetrating sclerokeratoplasty and frozen homologous corneoscleral grafting for the management of extensive canine epibulbar melanocytomas. Methods Medical records of canine patients treated at three different veterinary hospitals between 1999 and 2010 were reviewed. Signalment, location and extent of melanocytomas, recurrence rate, and early and late complications were reported. Patients were re‐examined postoperatively to provide follow‐up information. Results Patients included one intact male, three castrated males, six intact females, and one spayed female, with a median age of 5 years (range, 3–9). German Shepherds were overrepresented. Tumors extended from 2 to 4 clock hours at the limbus and up to 17 mm from clear cornea to globe equator. One case showed iridocorneal angle invasion; corneal involvement was present in all cases, and lipid keratopathy was present in four cases. In two cases, there was incomplete resection owing to tumor extent. Follow‐up time ranged from 3 to 72 months (median, 17 months), with one case of intraocular tumor progression. Early complications included anterior uveitis (11/11), intracameral fibrin (5/11), hyphema (4/11), corneal edema (4/11), exuberant corneal granulation tissue (2/11), focal retinal edema (1/11), dyscoria (1/11), and partial suture dehiscence (1/11). Late complications included corneal fibrosis and/or pigmentation (11/11), faint anterior cortical cataracts (3/11), and lipid keratopathy (1/11). Vision was retained in all cases. Conclusions This technique offers a surgically challenging but effective treatment for extensive epibulbar melanocytomas. In this case series, complications were mild and transient, with preservation of ocular anatomy and function.     

Unilateral invasive intraocular melanoma with choroidal and optic nerve involvement resulting in bilateral vision loss in a Labrador Retriever

A 10-year-old male castrated Labrador Retriever cross was referred for evaluation of acute vision loss. Ophthalmic examination revealed mild left sided exophthalmos, bilateral resting mydriasis, an absent direct and reduced consensual PLR in the left eye and reduced direct and absent consensual PLR in the right eye. Examination of the cornea and anterior segment with slit lamp biomicroscopy was unremarkable. Indirect fundoscopy revealed a left optic nerve head obscured by a darkly pigmented lesion. Fundic examination in the right eye was unremarkable. Magnetic resonance imaging revealed a smoothly marginated, lobulated cone to irregularly shaped, strongly T1 hyperintense, T2 and T2 fluid-attenuated inversion recovery hypointense, strongly contrast enhancing mass closely associated with the entire left optic nerve, extending across the optic chiasm and into the right optic nerve ventrally. Full clinical staging revealed no evidence of metastasis. Exenteration of the left eye was performed. Histopathology revealed an unencapsulated, poorly demarcated, multilobulated and infiltrative pigmented mass that was effacing the posterior choroid and optic nerve. The mass was composed of a moderately pleomorphic population of heavily pigmented polygonal cells arranged in sheets and clusters, displaying moderate anisocytosis and anisokaryosis. The population of cells contained moderate amounts of abundant brown-black granular pigment consistent with melanin within the cytoplasm. Mitotic figures averaged approximately three per ten 400× fields (2.37 mm²). This is the first report of a melanocytic tumor invading along the optic nerve and tract to result in contralateral vision loss.     

Chronic lameness caused by vascular compression of the iliac artery and vein by a malignant melanoma in the pelvis of a grey Spanish gelding

An 11-year-old grey Spanish gelding was presented with chronic lameness of the right hindlimb, with abduction of the limb during the swing phase. Based on a comprehensive lameness examination, the problem was localised to the pelvic region. Rectal and ultrasonographic examinations revealed a mass in the pelvic region in close association with the iliac artery and vein, and ultrasonographic indications of vascular compression. Considering the clinical presentation of a grey middle-aged horse with multifocal melanoma-like nodules at the level of the perineum, tail base, preputium and penis, a malignant melanoma with secondary vascular or neurological compression was suspected. Post-mortem and histological examination confirmed the diagnosis of malignant melanoma with vascular compression of the iliac artery and vein and infiltrative growth in the iliac vein.     

Prognostic value of somatic focal amplifications on chromosome 30 in canine oral melanoma

Canine oral melanoma is the first malignancy of the oral cavity in dogs and is characterized by a local invasiveness and a high metastatic propensity. A better knowledge of genetic alterations is expected to improve management of this tumour. Copy number alterations are known characteristics of mucosal melanomas both in dogs and humans. The goal of this study was to explore the prognostic value of somatic focal amplifications on chromosomes (Canis Familiaris [CFA]) 10 and 30 in canine oral melanoma. The cohort included 73 dogs with oral melanoma confirmed by histology, removed surgically without adjuvant therapy and with a minimal follow‐up of 6 months. Epidemiological, clinical and histological data were collected and quantitative‐PCR were performed on formalin‐fixed paraffin‐embedded (FFPE) samples to identify specific focal amplifications. The 73 dogs included in the study had a median survival time of 220 days. Focal amplifications on CFA 10 and 30 were recurrent (49.3% and 50.7% of cases, respectively) and CFA 30 amplification was significantly associated with the amelanotic phenotype (P = .046) and high mitotic index (MI; P = .0039). CFA 30 amplification was also linked to poor prognosis (P = .0005). Other negative prognostic factors included gingiva location (P = .003), lymphadenomegaly (P = .026), tumour ulceration at diagnosis (P = .003), MI superior to 6 mitoses over 10 fields (P = .001) and amelanotic tumour (P = .029). In multivariate analyses using Cox proportional hazards regression, CFA 30 amplification (Hazard ratio [HR] = 2.08; P = .011), tumour location (HR = 2.20; P = .005) and histological pigmentation (HR = 1.87; P = .036) were significantly associated with shorter survival time. Focal amplification of CFA 30 is linked to an aggressive subset and constitutes a new prognostic factor.     

Tumour‐infiltrating lymphocytes in canine melanocytic tumours: An investigation on the prognostic role of CD3+ and CD20+ lymphocytic populations

The study of the immune response in several types of tumours has been rapidly increasing in recent years with the dual aim of understanding the interactions between neoplastic and immune cells and their importance in cancer pathogenesis and progression, as well as identifying targets for cancer immunotherapy. Despite being considered one of the most immunogenic tumour types, melanoma can progress in the presence of abundant lymphocytic infiltration, therefore suggesting that the immune response is not able to efficiently control tumour growth. The purpose of this study was to investigate whether the density, distribution and grade of tumour‐infiltrating lymphocytes (TILs) in 97 canine melanocytic tumours is associated with histologic indicators of malignancy and can be considered a prognostic factor in the dog. As a further step in the characterization of the immune response in melanocytic tumours, an immunohistochemical investigation was performed to evaluate the two main populations of TILs, T‐lymphocytes (CD3⁺) and B‐lymphocytes (CD20⁺). The results of our study show that TILs are present in a large proportion of canine melanocytic tumours, especially in oral melanomas, and that the infiltrate is usually mild. The quantity of CD20⁺ TILs was significantly associated with some histologic prognostic factors, such as the mitotic count, the cellular pleomorphism and the percentage of pigmented cells. Remarkably, a high infiltration of CD20⁺ TILs was associated with tumour‐related death, presence of metastasis/recurrence, shorter overall and disease‐free survival, increased hazard of death and of developing recurrence/metastasis, hence representing a potential new negative prognostic factor in canine melanocytic tumours.     

The Marine Fungal Metabolite,Dicitrinone B,Induces A375 Cell Apoptosis through the ROS-Related Caspase Pathway

Dicitrinone B, a rare carbon-bridged citrinin dimer, was isolated from the marine-derived fungus, Penicillium citrinum. It was reported to have antitumor effects on tumor cells previously; however, the details of the mechanism remain unclear. In this study, we found that dicitrinone B inhibited the proliferation of multiple tumor types. Among them, the human malignant melanoma cell, A375, was confirmed to be the most sensitive. Morphologic evaluation, cell cycle arrest and apoptosis rate analysis results showed that dicitrinone B significantly induced A375 cell apoptosis. Subsequent observation of reactive oxygen species (ROS) accumulation and mitochondrial membrane potential (MMP) reduction revealed that the apoptosis induced by dicitrinone B may be triggered by over-producing ROS. Further studies indicated that the apoptosis was associated with both intrinsic and extrinsic apoptosis pathways under the regulation of Bcl-2 family proteins. Caspase-9, caspase-8 and caspase-3 were activated during the process, leading to PARP cleavage. The pan-caspase inhibitor, Z-VAD-FMK, could reverse dicitrinone B-induced apoptosis, suggesting that it is a caspase-dependent pathway. Our data for the first time showed that dicitrinone B inhibits the proliferation of tumor cells by inducing cell apoptosis. Moreover, compared with the first-line chemotherapy drug, 5-fluorouracil (5-Fu), dicitrinone B showed much more potent anticancer efficacy, suggesting that it might serve as a potential antitumor agent.     

杨梅枝条醇提物对A375细胞增殖和凋亡的影响及其分子机制

杨梅(Myrica rubra)具有较高的药用价值,利用杨梅枝条醇提物(Myrica rubra branch extract, MRBE)开展其对人恶性黑色素瘤A375细胞的作用研究。采用MTT法、PI流式细胞术、Annexin V-PI双染流式细胞术、实时荧光定量PCR(qRT-PCR)等方法,解析MRBE诱导A375细胞阻滞与凋亡的潜在作用机制。结果表明:MRBE对A375细胞的抑制率呈剂量依赖效应关系,抑制率最高达49.46%;流式细胞术结果表明,MRBE作用48 h可使A375细胞阻滞于S期并发生凋亡;qRT-PCR结果进一步表明,MRBE可能通过上调P21和下调CyclinA、CyclinD、CDK2、CDK4、CDK6、E2F1等基因的表达,诱导A375细胞阻滞于S期,同时下调Bcl-w和Raf基因的表达促使A375细胞凋亡。初步解析了MRBE对人恶性黑色素瘤A375细胞增殖和凋亡的作用机制,为利用MRBE研发抗黑色素瘤潜在药物提供科学依据,同时有助于提高杨梅枝条的药用附加值,拓宽杨梅产业链多元化发展。