PDF(Pubmed)
Abstract：
Canine oral melanoma is the most prevalent malignant tumor in dogs and has a poor prognosis due to its high aggressiveness and high metastasis and recurrence rates. More research is needed into its treatment and to understand its pathogenic factors. In this study, we isolated a canine oral mucosal melanoma (COMM) cell line designated as COMM6605, which has now been stably passaged for more than 100 generations, with a successful monoclonal assay and a cell multiplication time of 22.2 h. G-banded karyotype analysis of the COMM6605 cell line revealed an abnormal chromosome count ranging from 45 to 74, with the identification of a double-armed chromosome as the characteristic marker chromosome of this cell line. The oral intralingual and dorsal subcutaneous implantation models of BALB/c-nu mice were successfully established; Melan-A (MLANA), S100 beta protein (S100β), PNL2, tyrosinase-related protein 1 (TRP1), and tyrosinase-related protein 2 (TRP2) were stably expressed positively in the canine oral tumor sections, tumor cell lines, and tumor sections of tumor-bearing mice. Sublines COMM6605-Luc-EGFP and COMM6605-Cherry were established through lentiviral transfection, with COMM6605-Luc-EGFP co-expressing firefly luciferase (Luc) and enhanced green fluorescent protein (EGFP) and COMM6605-Cherry expressing the Cherry fluorescent protein gene. The COMM6605-Luc-EGFP fluorescent cell subline was injected via the tail vein and caused lung and lymph node metastasis, as detected by mouse live imaging, which can be used as an animal model to simulate the latter steps of hematogenous spread during tumor metastasis. The canine oral melanoma cell line COMM6605 and two sublines isolated and characterized in this study can offer a valuable model for studying mucosal melanoma.
摘要：
犬口腔黑色素瘤是犬中最常见的恶性肿瘤，由于其高侵袭性，高转移和复发率，预后较差。需要对其治疗进行更多研究并了解其致病因素。在这项研究中,我们分离了犬口腔粘膜黑色素瘤（COMM）细胞系，命名为COMM6605，现在已经稳定传代超过100代，成功的单克隆测定和22.2h的细胞增殖时间。COMM6605细胞系的G带核型分析显示，染色体计数异常，范围为45至74，鉴定出双臂染色体是该细胞系的特征性标记染色体。成功建立BALB/c-nu小鼠口腔舌内和背侧皮下植入模型;Melan-A(MLANA),S100β蛋白(S100β),PNL2,酪氨酸酶相关蛋白1(TRP1),酪氨酸酶相关蛋白2(TRP2)在犬口腔肿瘤切片中稳定表达,肿瘤细胞系，和荷瘤小鼠的肿瘤切片。SublinesCOMM6605-Luc-EGFP和COMM6605-Cherry通过慢病毒转染建立，与COMM6605-Luc-EGFP共表达萤火虫荧光素酶（Luc）和增强型绿色荧光蛋白（EGFP）和COMM6605-Cherry共表达樱桃荧光蛋白基因。经尾静脉注射COMM6605-Luc-EGFP荧光细胞亚系，引起肺和淋巴结转移，通过鼠标实时成像检测到，可以用作动物模型来模拟肿瘤转移过程中血行扩散的后期步骤。在本研究中分离和表征的犬口腔黑素瘤细胞系COMM6605和两个亚系可以为研究粘膜黑素瘤提供有价值的模型。
