绿色荧光蛋白标记的恶性组织细胞增生症瘤细胞眼前房移植模型  

A new malignant histiocytosis expressing green fluorescent protein model in the anterior chamber

作　　者：聂莉[1] 李永平[1] 李亮平 张文忻[1] 林健贤[1] 张波[1] 

机构地区：[1]中山大学中山眼科中心研究所 [2]Max-Delbruck-Center for Molecular Medicine,13092 Berlin,Germany

出　　处：《眼科研究》2006年第2期152-155,共4页Chinese Ophthalmic Research

基　　金：广东省科技计划项目资助(2003A3020302)

摘　　要：目的建立绿色荧光蛋白(GFP)标记的恶性组织细胞增生症瘤细胞cy15在眼前房生长及浸润的模型,探讨cy15细胞在前房生长的规律及GFP标记活细胞的优势。方法将带有GFP的逆转录病毒MP71-GFP-PRE转入cy15瘤细胞,获得稳定表达GFP的cy15GFP细胞系。20只BALB/C小鼠(20只眼)每只眼前房注入2μl细胞密度为2×106个/ml的cy15GFP瘤细胞悬液,术后分别在裂隙灯和荧光显微镜下观察瘤细胞在前房的生长情况,连续观察30d,分别在瘤细胞植入前房后第15d、20d、30d时处死小鼠做眼球HE病理切片。结果成功地建立了稳定表达GFP的cy15GFP细胞系。接种的20只BALB/c鼠眼前房均可见肿瘤细胞生长,借助于荧光显微镜能动态地观察到肿瘤细胞在活体小鼠眼前房的生长浸润过程。结论建立的GFP标记的肿瘤细胞眼前房接种模型为研究眼内肿瘤细胞生长与浸润的规律以及抗肿瘤药物的筛选提供了一种新的手段。Objective The ocular invasion of malignant histiocytosis is often misdiagnosed in clinic. Our research was to observe the growth and invasion rule of malignant histiocytosis cells cy15 expressing green fluorescent protein （GFP） inoculated in the anterior chamber of BALB/C mice and the superiority of GFP as the marker of living cell. Methods MP71-GFP-PRE, a retrovirus vector expressing GFP, was transferred into malignant histiocytosis cy15 cell line. The cell clones expressing GFP steadily were selected by neomycin and fluorescence microscope. 2 μl of cy15GFP cells （density at 2.0×10^6 cells/ml） were injected into the anterior chamber of 20 BALB/c mice under operation microscope. The growth of cy15 in the anterior chamber was observed with fluorescence microscope and slit lamp. Results Malignant histiocytosis cy15 GFP cells expressed GFP steadily and developed tumor in the anterior chamber. At the early stage, single tumor cell was observed in the anterior chamber under fluorescence microscope and cloned to generate green fluorescence mass as time lapse. Pathologic section also disclosed the tumor cells in the anterior chamber. Conclusion The growth process of tumor cells expressing GFP in the anterior chamber of living mice is observed. This model is a good experimental animal model in the studies on ocular tumor growth, tumor invasion mechanism and drug selection.

关 键 词：恶性组织细胞增生症 绿色荧光蛋白 眼 前房 

分 类 号：R733.1[医药卫生—肿瘤]