Model of orthotopic transplatation tumor of malignant choroidal melanoma labeled by green fluorescent protein
- VernacularTitle:绿色荧光蛋白标记的脉络膜恶性黑色素瘤原位移植瘤模型
- Author:
Xiaoli WANG
;
Ping XU
;
Feng WANG
- Publication Type:Journal Article
- Keywords:
Melanoma, experimental;
Eye neoplasms;
Neoplasm metastasis;
Fluorescent antibody technique
- From:
Chinese Journal of Ocular Fundus Diseases
2000;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a new model of orthotopic-transplatation tumor of human malignant choroidal melanoma. Methods pEGFP-N1, the eukaryotic expressive plasmid of green fluorescent protein (GFP), was transfered into human malignant choroidal melanoma cell line (OCM-1) by liposome lipofectanine, then the cell clones with stable GFP expression were screened out by means of neomycin, fluorescence microscope, and flow cytometer. Two ?l cell suspension of OCM-1 cells with GFP expression with the density of 4.5?107-5.5?107 cells/ml was injected into the subretinal space of right eyes of 40 nude mice (40 eyes) under binocular operating microscope with left eyes as the control ones. The growth of the transplanted malignant choroidal melanoma was observed in vivo using the fluorescence stereomicroscope. The mice were killed at different time after the operation to observe the metastasis of the tumor to optic nerve, brain and other organs including lung, liver, kidney and spleen. Moreover, pathological detection and immunohistochemical staining of GFP was carried out. Results At the postoperative 10 th-12 th days, the growths of the transplanted malignant choroidal melanoma with dilated and distorted blood vessels and neovascularization were observed; at the postoperative 20 th-22 nd days, the melanoma occupied the whole cavity of vitreous body; and at the postoperative 24 th-26 th days, the transplanted tumor grew out of the eye. Metastases of the carcinoma to olfactory bulb, kidney, lung and liver were seen at the failure phase soon after the extra-ocular phase. The histopathological characteristics of the transplanted tumor were similar to those of human, and the results of immunohistochemical staining showed positive expression of GFP in the tumor cells. Conclusion The orthotopic model of malignant choroidal melanoma set up via injection of human malignant choroidal melanoma cells labeled by GFP into the subretinal space of nude mice may provide a new approach to investigate the natural courses of growth and metastasis of malignant choroidal melanoma.
