Generation of clonal zebrafish lines and transplantable hepatic tumors
Generation of clonal zebrafish lines and transplantable hepatic tumors
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DOI：
10.1038/nprot.2010.8
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发表时间：
2010-01-01
期刊：
影响因子：
14.8
通讯作者：
Revskoy, Sergei
中科院分区：
文献类型：
--
作者：
Mizgirev, Igor;Revskoy, Sergei
在啮齿类动物的癌症研究中，可移植肿瘤是公认的黄金标准。由于缺乏真正的近交系，这种模型在斑马鱼中的进展一直受到限制。我们已经产生了几种纯合子二倍体克隆斑马鱼系，允许肿瘤细胞从一条鱼连续移植到另一条鱼，而不需要亚致死γ辐射。在近交克隆斑马鱼系中最初诱导和维持的可移植肿瘤的光谱仅限于不同类型的自发和二乙基亚硝胺诱导的肝脏肿瘤。然而，该模型可以很容易地扩展到广泛的肝外肿瘤，具有明确诱导机制的转基因肿瘤和荧光标记肿瘤系。这些模型将通过体内成像进一步促进对侵袭性肿瘤生长、血管生成、转移和肿瘤启动细胞的深入分析，并为高通量（HTP）筛选抗癌治疗药物（包括生物反应调节剂）提供一个具有成本效益的系统。此外，纯合子斑马鱼系是免疫遗传学、定量性状位点定位和其他遗传应用不可缺少的工具。整个过程，从雌性纯合子鱼（奠基者）的产生到获得3-4个连续的同基因肿瘤，大约需要12-18个月。这个时间范围很大程度上取决于肿瘤诱导的方法、肿瘤类型和肿瘤生长速度。
Transplantable tumors are an accepted gold standard in cancer studies in rodents. The progress of this model in zebrafish has long been constrained by the lack of true inbred lines in zebrafish. We have generated several lines of homozygous diploid clonal zebrafish lines, which allow serial transplantations of tumor cells from one fish to another without sublethal gamma-irradiation. The spectrum of transplantable tumors that were initially induced and maintained in inbred clonal zebrafish lines was limited to different types of spontaneous and diethylnitrosamine-induced hepatic tumors. However, this model can readily be extended to a broad range of extrahepatic tumors, transgenic tumors with defined mechanisms of induction and fluorescence-tagged tumor lines. These models will further facilitate in-depth analysis of invasive tumor growth, angiogenesis, metastasis and tumor-initiating cells by in vivo imaging and provide a cost-effective system for high-throughput (HTP) screening of anticancer therapeutics, including biological response modifiers. In addition, homozygous zebrafish lines are an indispensable tool for immunogenetics, mapping of quantitative trait loci and other genetic applications. The whole procedure, from generation of a gynogenetic female homozygous fish (a founder) to obtaining 3-4 consecutive passages of a syngeneic tumor, takes similar to 12-18 months. This time-frame largely depends on methods of tumor induction, tumor type and tumor growth rate.