Mammalian tumor xenografts induce neovascularization in zebrafish embryos
Mammalian tumor xenografts induce neovascularization in zebrafish embryos
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DOI：
10.1158/0008-5472.can-06-4268
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发表时间：
2007-04-01
期刊：
影响因子：
11.2
通讯作者：
Presta, Marco
中科院分区：
文献类型：
--
作者：
Nicoli, Stefania;Ribatti, Domenico;Presta, Marco
斑马鱼(Danio Rerio)/肿瘤异种移植模型代表了一种强大的肿瘤新模型系统。在这里，我们描述了一种新的利用斑马鱼模型来研究肿瘤血管生成的方法，肿瘤血管生成是癌症进展的关键步骤，也是抗肿瘤治疗的靶点。表达血管生成成纤维细胞生长因子2和/或血管内皮生长因子的人和小鼠肿瘤细胞系在受精后48h被移植到斑马鱼胚胎发育中的肠下血管附近，诱导迅速形成新的微血管。相反，在产生这些血管生成生长因子的过程中存在缺陷的相关细胞克隆不会产生血管生成反应。新形成的血管从斑马鱼胚胎的肠下神经丛中萌发，穿透肿瘤移植物，并表达早期内皮标记物Fli-1、%TEGF受体-2(AIEGFR2/KDR)和VE-cadherin的转录本。因此，当肿瘤细胞被注射到VEGFR2/KDR启动子控制下表达绿色荧光蛋白的转基因斑马鱼胚胎中时，绿色荧光蛋白阳性的新生血管渗透到移植物中。在肿瘤细胞注射后立即系统地将斑马鱼胚胎暴露于原型抗血管生成抑制剂，包括成纤维细胞生长因子受体酪氨酸激酶抑制剂SU5402和‘%rEGFR2/KDR酪氨酸激酶抑制剂SU5416，在不影响正常血管发育的情况下抑制肿瘤诱导的血管生成。因此，注射反义吗啉寡核苷酸使VE-钙粘蛋白基因失活可以抑制肿瘤新生血管，而不影响节段间和肠段下血管的发育。这些数据表明，斑马鱼/肿瘤异种移植模型是研究肿瘤血管新生过程的一种新工具，可用于肿瘤血管生成的药物发现和基因靶向。
The zebrafish (Danio rerio)/tumor xenograft model represents a powerful new model system in cancer. Here, we describe a novel exploitation of the zebrafish model to investigate tumor angiogenesis, a pivotal step in cancer progression and target for antitumor therapies. Human and murine tumor cell lines that express the angiogenic fibroblast growth factor (FGF) 2 and/or vascular endothelial growth factor (VEGF) induce the rapid formation of a new microvasculature when grafted close to the developing subintestinal vessels of zebrafish embryos at 48 h postfertilization. Instead, no angiogenic response was exerted by related cell clones defective in the production of these angiogenic growth factors. The newly formed blood vessels sprout from the subintestinal plexus of the zebrafish embryo, penetrate the tumor graft, and express the transcripts for the zebrafish orthologues of the early endothelial markers Fli-1, %TEGF receptor-2 (AIEGFR2/KDR), and VE-cadherin. Accordingly, green fluorescent protein-positive neovessels infiltrate the graft when tumor cells are injected in transgenic VEGFR2.G-RCFP zebrafish embryos that express green fluorescent protein under the control of the VEGFR2/KDR promoter. Systemic exposure of zebrafish embryos immediately after tumor cell injection to prototypic antiangiogenic inhibitors, including the FGF receptor tyrosine kinase inhibitor SU5402 and the '%rEGFR2/KDR tyrosine kinase inhibitor SU5416, suppresses tumor-induced angiogenesis without affecting normal blood vessel development. Accordingly, VE-cadherin gene inactivation by antisense morpholino oligonucleotide injection inhibits tumor neovascularization without affecting the development of intersegmental and subintestinal vessels. These data show that the zebrafish/ tumor xenograft model represents a novel tool for investigating the neovascularization process exploitable for drug discovery and gene targeting in tumor angiogenesis.