[1]王祯莲,金畅.GFP标记的人葡萄膜黑色素瘤裸鼠模型的构建[J].常州大学学报(自然科学版),2016,(05):78-81,86.[doi:10.3969/j.issn.2095-0411.2016.05.014]
　WANG Zhenlian,JIN Chang.Establishment of an Experimental Animal Model of Uveal Melanoma in Mouse Engineered by Green Fluorescent Protein[J].Journal of Changzhou University(Natural Science Edition),2016,(05):78-81,86.[doi:10.3969/j.issn.2095-0411.2016.05.014]
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GFP标记的人葡萄膜黑色素瘤裸鼠模型的构建()
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常州大学学报(自然科学版)[ISSN:2095-0411/CN:32-1822/N]

卷:
期数:
2016年05期
页码:
78-81,86
栏目:
生物医学工程
出版日期:
2016-09-30

文章信息/Info

Title:
Establishment of an Experimental Animal Model of Uveal Melanoma in Mouse Engineered by Green Fluorescent Protein
作者:
王祯莲1; 金畅2
1.常州大学 护理学院,江苏 常州213164; 2.温州医科大学 眼视光学院,浙江 温州325003
Author(s):
WANG Zhenlian1;  JIN Chang2
1. School of Nursing,Changzhou University,Changzhou 213164,China; 2. School of Ophthalmology and Optometry, Wenzhou Medical College, Wenzhou 325003,China
关键词:
葡萄膜黑色素瘤;  绿色荧光蛋白;  动物模型
Keywords:
uveal melanoma;  GFP;  animal model
分类号:
R 739.7
DOI:
10.3969/j.issn.2095-0411.2016.05.014
文献标志码:
A
摘要:
探讨通过构建稳定表达GFP的人葡萄膜黑色素瘤细胞系,建立新型人葡萄膜黑色素瘤裸鼠皮下移植模型。培养人葡萄膜黑色素瘤细胞系M17和M23,收集细胞并制备细胞悬液,每只裸鼠于腹部皮下注射细胞悬液0.1 mL约2×106个细胞,观察M17和M23细胞系成瘤率、成瘤时间、肿瘤生长、转移情况及组织病理学形态。用绿色荧光蛋白(GFP)表达质粒转染M17和M23细胞,筛选稳定克隆。在荧光显微镜下观察GFP转染阳性率。将稳定克隆腹部皮下注射裸鼠,接种6周后处死裸鼠,取新鲜肿瘤组织块做病理切片,免疫组化观察组织块荧光表达情况。实验发现裸鼠成瘤率100%,病理切片检查证实为黑色素瘤。成瘤时间为2周左右,其中一只裸鼠于接种4周后出现腹水、肝内转移。GFP转染率达90%以上。裸鼠肿瘤新鲜组织块发出绿色荧光,而包围瘤块的正常皮肤没有荧光。以GFP基因标记的葡萄膜黑色素瘤细胞经裸鼠腹部皮下移植建立的葡萄膜黑色素瘤模型,为研究自然状态下肿瘤的生长和转移提供了一个简单、可靠的方法。
Abstract:
To establish a novel human uveal melanoma mouse model methods, human uveal melanoma cell lines M17 and M23 were cultured in vitro. Aliquots of 2×106 cells per 0.1 mL suspension was subcutaneously transplanted in the abdomen of BALB/C nude mice. The growth condition of implanted tumors and the metastases were investigated. pEGFP-N1 was transferred into M17 and M23 cell lines by liposome lipofectamine, the cell clones with stable GFP expression were screened out by means of G418 and fluorescence microscope. GFP positive M17 or M23 cells were subcutaneously injected into nude mice to establish uveal melanoma animal models. All the animals were sacrificed after 6 weeks. Pathological detection and immunohistochemical staining of GFP was carried out. The rates of tumor induction were 100%. At 4 weeks, histologic examination showed that one mouse formed hepatic micrometastases. 90% of uveal melanoma cells stably expressed EGFP. GFP-labeled tumor cells displayed green fluorescence under fluorescent microscopy. The model of uveal melanoma set up via injection of human uveal melanoma cells labeled by GFP into the subcutaneous of nude mice may provide a new approach to investigate the natural courses of growth and metastasis of malignant uveal melanoma.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2016-01-23。基金项目:国家自然科学基金青年基金项目(81201657)。作者简介:王祯莲(1980—),女,山西阳泉人,硕士,讲师,主要从事肿瘤侵袭转移研究。
更新日期/Last Update: 2016-10-10