Ren Jiang, Liu Sijia, Cui Chao, Ten Dijke Peter
Department of Molecular Cell Biology, Cancer Genomics Centre Netherlands, Leiden University Medical Center.
Department of Molecular Cell Biology, Cancer Genomics Centre Netherlands, Leiden University Medical Center;
J Vis Exp. 2017 Apr 25(122):55459. doi: 10.3791/55459.
In many cases, cancer patients do not die of a primary tumor, but rather because of metastasis. Although numerous rodent models are available for studying cancer metastasis in vivo, other efficient, reliable, low-cost models are needed to quickly access the potential effects of (epi)genetic changes or pharmacological compounds. As such, we illustrate and explain the feasibility of xenograft models using human breast cancer cells injected into zebrafish embryos to support this goal. Under the microscope, fluorescent proteins or chemically labeled human breast cancer cells are transplanted into transgenic zebrafish embryos, Tg (fli:EGFP), at the perivitelline space or duct of Cuvier (Doc) 48 h after fertilization. Shortly afterwards, the temporal-spatial process of cancer cell invasion, dissemination, and metastasis in the living fish body is visualized under a fluorescent microscope. The models using different injection sites, i.e., perivitelline space or Doc are complementary to one another, reflecting the early stage (intravasation step) and late stage (extravasation step) of the multistep metastatic cascade of events. Moreover, peritumoral and intratumoral angiogenesis can be observed with the injection into the perivitelline space. The entire experimental period is no more than 8 days. These two models combine cell labeling, micro-transplantation, and fluorescence imaging techniques, enabling the rapid evaluation of cancer metastasis in response to genetic and pharmacological manipulations.
在许多情况下，癌症患者并非死于原发性肿瘤，而是死于转移。尽管有许多啮齿动物模型可用于在体内研究癌症转移，但仍需要其他高效、可靠且低成本的模型，以便快速了解（表观）遗传变化或药理化合物的潜在作用。因此，我们阐述并解释了将人乳腺癌细胞注射到斑马鱼胚胎中建立异种移植模型的可行性，以支持这一目标。在显微镜下，将荧光蛋白或化学标记的人乳腺癌细胞在受精后48小时移植到转基因斑马鱼胚胎Tg (fli:EGFP) 的卵黄周隙或居维叶氏管（Doc）中。此后不久，在荧光显微镜下可观察到癌细胞在活体鱼体内侵袭、播散和转移的时空过程。使用不同注射部位（即卵黄周隙或Doc）的模型相互补充，反映了多步骤转移级联事件的早期阶段（血管内渗步骤）和晚期阶段（血管外渗步骤）。此外，向卵黄周隙注射可观察到肿瘤周围和肿瘤内的血管生成。整个实验周期不超过8天。这两种模型结合了细胞标记、微移植和荧光成像技术，能够快速评估癌症转移对基因和药物操作的反应。
