Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Washington, Washington, DC, USA.
Methods Mol Biol. 2021;2294:3-16. doi: 10.1007/978-1-0716-1350-4_1.
Cancer cell vascular invasion and extravasation at metastatic sites are hallmarks of malignant progression of cancer and associated with poor disease outcome. Here we describe an in vivo approach to study the invasive ability of cancer cells into the vasculature and their hematogenous metastatic seeding in zebrafish (Danio rerio). In one approach, extravasation of fluorescently labeled cancer cells is monitored in zebrafish embryos whose vasculature is marked by a contrasting fluorescent reporter. After injection into the precardiac sinus of 2-day-old embryos, cancer cells can extravasate from the vasculature into tissues over the next few days. Extravasated cancer cells are identified and counted in live embryos via fluorescence microscopy. In a second approach, intravasation of cancer cells can be evaluated by changing their injection site to the yolk sac of zebrafish embryos. In addition to monitoring the impact of drivers of malignant progression, candidate inhibitors can be studied in this in vivo model system for their efficacy as well as their toxicity for the host.
癌细胞在转移部位的血管侵犯和外渗是癌症恶性进展的标志，与不良的疾病结局相关。在这里，我们描述了一种在活体斑马鱼（Danio rerio）中研究癌细胞进入血管的侵袭能力及其血源性转移定植的方法。在一种方法中，通过荧光标记的癌细胞的外渗来监测斑马鱼胚胎的血管，这些胚胎的血管由对比荧光报告基因标记。在 2 日龄胚胎的心前窦注射后，癌细胞可以在接下来的几天内从血管渗出到组织中。通过荧光显微镜在活体胚胎中识别和计数外渗的癌细胞。在第二种方法中，通过将癌细胞的注射部位改为斑马鱼胚胎的卵黄囊，可以评估癌细胞的血管内渗。除了监测恶性进展驱动因素的影响外，还可以在这种体内模型系统中研究候选抑制剂的疗效及其对宿主的毒性。
