一种体内实时观察黑色素瘤的斑马鱼模型

doi:10.3969/j.issn.1674-5817.2018.01.004

实验动物与比较医学 ›› 2018, Vol. 38 ›› Issue (1): 22-28.DOI: 10.3969/j.issn.1674-5817.2018.01.004

所属专题：实验动物资源开发与利用

一种体内实时观察黑色素瘤的斑马鱼模型

周斌¹, 盛哲津², 冯琛卓³, 李利妹⁴

1.同济大学附属东方医院血管外科,上海 200120;
2.同济大学生命科学与技术学院,上海 200092;
3.杭州医学院基础医学部,杭州 310053;
4.同济大学附属东方医院医学转化中心,上海 200120

收稿日期:2017-08-28 出版日期:2018-02-25 发布日期:2021-03-01
作者简介:周斌(1983-),男,博士,主治医师。E-mail:mcdropin@126.com
基金资助:
中央高校基础科研业务专项基金(No.1507219069),浙江省自然科学基金一般项目(No.LY18H090009),浦东新区卫生计生委科技项目(No.PW2017B-7)

A Melanoma Zebrafish Model for Real-time Imaging in vivo

ZHOU Bin¹, SHENG Zhe-jin², FENG Chen-zhuo³, LI Li-mei⁴

1. Department of Vascular Surgery,Shanghai East Hospital,School of Medicine,Tongji University,Shanghai 200120,China;
2. School of Life Science and Technology,Tongji University,Shanghai 200092,China;
3. Department of Basic Medical Sciences,Hangzhou Medical College,Hangzhou 310053,China;
4. Research Center for Translational Medicine,Shanghai East Hospital,School of Medicine,Tongji University,Shanghai 200120,China

Received:2017-08-28 Online:2018-02-25 Published:2021-03-01

摘要/Abstract

摘要： 目的建立一种在体内实时观察黑色素瘤发生发展的斑马鱼模型, 为黑色素瘤的相关研究提供一个便捷的活体模型。方法构建红色荧光蛋白标记的黑色素瘤表达系统pMITFa1-RedV12/hRas, 利用小鼠B16黑色素细胞验证其表达效果。再将pMITFa1-RedV12/hRas载体导入1-细胞期的斑马鱼受精卵中, 采用活体成像系统实时观察黑色素瘤在斑马鱼体内的形成过程。结果 pMITFa1-RedV12/hRas载体在B16细胞中成功表达。显微注射pMITFa1-RedV12/hRas载体到1-cell期的斑马鱼受精卵3 d后,在荧光显微镜下观察到荧光标记的黑色素瘤;8周后,肉眼可以明显观察到黑色素瘤。黑色瘤的成瘤率达100%。pMITFa1-RedV12/hRas载体被显微注射到TG(zlyz:EGFP)转基因斑马鱼的受精卵中后,在活体成像系统下清晰地观察到红色荧光标记的黑色素瘤细胞与绿色荧光标记的巨噬细胞的相互作用。结论成功构建了经济简便、易操作的在体内实时观察黑色素瘤的斑马鱼模型。

关键词: 黑色素瘤模型, 斑马鱼, 实时观察

Abstract: Objective To establish a zebrafish model for real-time monotoring the development of melanoma in vivo and screening relative drugs.Methods The expression effect of pMITFa1-RedV12/hRas plasmid,in which the melanoma was labeled by red fluorescent protein (RFP),was verified in mouse B16 melanocytes.After microinjecting pMITFa1-RedV12/hRas into 1- cell stage of fertilized eggs of zebrafish,we monitored the process of melanoma formation with in vivo imaging system in zebrafish.Results pMITFa1-RedV12/hRas was successfully expressed in mouse melanoma B16 cells.Three days after microinjection of pMITFa1-RedV12/hRas vector to 1-cell satge of zebrafish,melanomas labeled by RFP protein was observed under the fluorescence microscope.Eight weeks later,melanomas were observed clearly by eyes.The rate of tumor formation was 100%.When microinjected into pMITFa1-RedV12/hRas vector into fertilized eggs of TG(zlyz:EGFP) transgenic zebrafish,the interaction between the melanoma cells labeled by RFP protein and the macrophages labeled by green fluorescent protein was clearly observed in in vivo imaging system.Conclusion Successfully established a zebrafish model for real-time monitoring melanoma in vivo,which is cheap and easy to operate.

Key words: Melanoma model, Zebrafish, Real-time imaging

中图分类号:

Q95-33

周斌, 盛哲津, 冯琛卓, 李利妹. 一种体内实时观察黑色素瘤的斑马鱼模型[J]. 实验动物与比较医学, 2018, 38(1): 22-28.

ZHOU Bin, SHENG Zhe-jin, FENG Chen-zhuo, LI Li-mei. A Melanoma Zebrafish Model for Real-time Imaging in vivo[J]. Laboratory Animal and Comparative Medicine, 2018, 38(1): 22-28.

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一种体内实时观察黑色素瘤的斑马鱼模型

A Melanoma Zebrafish Model for Real-time Imaging in vivo

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