兔葡萄膜黑色素瘤模型建立方法的实验研究

Establishment of an experimental animal model of uveal melanoma in rabbits eyes

目的建立兔眼脉络膜黑色素瘤动物模型,观察肿瘤的生长特性。设计实验研究。研究对象新西兰大白兔20只(20眼)。方法将培养的B16F10鼠皮肤黑色素瘤细胞悬液种植到C57BL/6黑鼠大腿内侧皮下,7～12天孵育成直径10mm团块后制成肿瘤碎片,种植到新西兰大白兔右眼脉络膜上腔,自种植之日起予环孢霉素A20mg/kg肌肉注射以抑制淋巴细胞免疫功能。每周行间接检眼镜、眼底照相、B超、彩色超声多普勒观察并记录兔眼底肿瘤大小、形状、眼部其他组织的改变以及肿瘤内部血液供应情况。观察6周后眼球摘除,行HE染色组织病理学检查。主要指标肿瘤最大基底径、高度、细胞类型、肿瘤内血管分布情况以及眼部其他组织肿瘤浸润情况。结果除1只(5%)兔因呕吐物窒息死亡外,余19只兔19眼(95%)肿瘤存活并迅速生长。其中1眼(5%)自种植4周开始肿瘤自行萎缩,第6周12眼(60%)肿瘤充满玻璃体腔。2周、3周、4周B超测量肿瘤平均最大基底径分别为3.1mm、6.1mm、12.2mm,平均高度分别为2.2mm、2.7mm、8.5mm。HE染色示肿瘤以梭形细胞为主,排列规则紧密,部分坏死,血管充盈。18眼(90%)均可见眼部其他组织受肿瘤细胞浸润,3眼(15%)肿瘤突破巩膜向球外蔓延。结论 B16F10鼠皮肤黑色素瘤细胞株兔眼脉络膜上腔种植后肿瘤迅速生长,模型建造成功率高。

Objective To establish an experimental xenograft animal model of uveal melanoma, and evaluate the feasibility. Design Experimental study. Participants 20 eyes of 20 New Zealand albino rabbits. Methods B16FI0 melanoma cell line suspension was subcutaneously injected in C57BL/6 black rat. Fragments of tumor was implanted into the suprachoroidal space of New Zealand rabbits, when tumor diameter exceeded 10 mm after 7 to 12 days. Cyclosponine A was injected everyday to suppress rabbit immune system. Funduscopies, B ultrasonography, corlor Doppler imaging were applied once a week. All the animals were sacrificed after 6 weeks, followed with histopathological examination of eyeballs. Main Outcome Measures Tumour diameter of the largest direction of base, tumor height, cell type, vascular distribution within tumor, tumor invasion. Results B16F10 cell line growed rapidly in 19 eyes （95%）, 60% of which filled most of the vitreous cavity within 6 weeks. Mean largest tumor diameter measured by B uhrasonography of 2 weeks, 3 weeks and 4 weeks was 3.1 ram, 6.1 ram, 12.2 mm respetively, and mean height was 2.2 mm, 2.7 mm, 8.5 mm respetively. Extrocular invasion was found in 3 eyes （15%）. Conclusion It is feasible to establish xenograft animal model of uveal melanoma with B16F10 melanoma cell line. （Ophthalmol CHN, 2010, 19： 130-134）