BAMBI基因过表达人结肠癌细胞移植斑马鱼致肝转移模型的构建

发布时间：2018-04-27 10:59

本文选题：结肠肿瘤 + 肝转移　；参考：《中华临床医师杂志(电子版)》2015年21期

【摘要】：目的构建BAMBI基因过表达人结肠癌细胞移植斑马鱼肝转移模型,分析斑马鱼肝内转移性结肠癌细胞BAMBI基因表达水平与肝转移发生率的关联性。方法分别构建BAMBI过表达稳转结肠癌细胞株SW620(BAMBI+组)、空载体红色荧光蛋白基因转染结肠癌细胞株SW620(EV组),分别显微注射到受精后第2天转基因斑马鱼Tg(Apo14∶GFP)胚胎卵黄囊(每组各680条),激光共聚焦荧光显微镜观察、拍摄,采用Image J软件测量转移至斑马鱼肝脏内结肠癌细胞红色荧光表达强度,计算肝转移发生率;采用t检验比较BAMBI+组及EV组斑马鱼肝脏内结肠癌细胞红色荧光表达强度及肝转移发生率的差异。结果与EV组相比,激光共聚焦荧光显微观察显示BAMBI+组斑马鱼肝内结肠癌细胞红色荧光表达量显著增加(t=6.247,P=0.003 3);BAMBI+组斑马鱼肝转移发生率与EV组相比显著增加(t=4.276,P=0.012 9)。结论构建了基因修饰人结肠癌细胞移植斑马鱼致肝转移模型,可较好地模拟活体状态的结肠癌肝转移;BAMBI基因可促进异种移植入斑马鱼的结肠癌细胞发生肝转移。
[Abstract]:Objective to establish a model of liver metastasis in zebrafish transplanted with human colon cancer cells with BAMBI gene overexpression, and to analyze the relationship between the expression of BAMBI gene and the incidence of liver metastasis in metastatic colon cancer cells of zebrafish. Methods BAMBI overexpression in colon cancer cell line SW620(BAMBI group was constructed, and empty vector red fluorescent protein gene was transfected into colon cancer cell line SW620(EV group. The embryo yolk sac of transgenic zebrafish TgApo14: GFP was microinjected into the embryo on the second day after fertilization. (680 pieces of each group were observed by laser confocal fluorescence microscope) Image J software was used to measure the expression intensity of red fluorescence in colon cancer cells transferred to zebrafish liver, and the incidence of liver metastasis was calculated. T test was used to compare the expression intensity of red fluorescence and the incidence of liver metastasis in BAMBI group and EV group. Results compared with EV group, laser confocal fluorescence microscopy showed that the expression of red fluorescence in colon cancer cells of zebrafish in BAMBI group was significantly higher than that in EV group. The incidence of hepatic metastasis of zebrafish in BAMBI group was significantly higher than that in EV group. Conclusion the model of liver metastasis induced by gene modified human colon cancer cell transplantation in zebrafish is constructed, which can simulate the liver metastasis of colon cancer cells in vivo and can promote liver metastasis of colonic cancer cells transplanted into zebrafish xenografts.
【作者单位】：湖州师范学院附属第一医院中心实验室;浙江大学医学院基础医学系细胞生物学教研室;湖州师范学院医学院病理学教研室;中国科学院水生生物研究所鱼类生物学及渔业生物技术研究中心鱼类发育遗传学与细胞工程学学科组;
【基金】：浙江省公益性技术应用研究计划项目(实验动物平台项目)(2013C37031) 湖州市科技计划项目(公益应用技术重点)(2013GZ14)
【分类号】：R735.3;R-332

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