Oncode Institute and Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands.
Institute of Biology, Leiden University, Leiden, The Netherlands.
Methods Mol Biol. 2022;2488:67-80. doi: 10.1007/978-1-0716-2277-3_6.
Transforming growth factor-β (TGF-β) family members have pivotal functions in controlling breast cancer progression, acting not only on cancer cells but also on other cells within the tumor microenvironment. Here we describe embryonic zebrafish xenograft assays to investigate how TGF-β family signaling controls breast cancer cell intravasation, extravasation and regulates tumor angiogenesis. Fluorescently mCherry-labeled breast cancer cells are injected in the perivitelline space or Duct of Cuvier of Tg (fli:EGFP) transgenic Casper zebrafish embryos, in which the zebrafish express enhanced green fluorescent protein in the entire vasculature. The dynamic responses of migratory and invasive human cancer cells, and the induction of new blood vessel formation by the cancer cells in zebrafish host, are visualized using a fluorescent microscope. These assays provide efficient, reliable, low-cost models to investigate the effect of (epi)genetic modulators and pharmacological compounds that perturb the activity of TGF-β family signaling components on breast cancer cell metastasis and angiogenesis.
转化生长因子-β（TGF-β）家族成员在控制乳腺癌进展方面具有关键作用，不仅作用于癌细胞，还作用于肿瘤微环境中的其他细胞。在这里，我们描述了胚胎斑马鱼异种移植实验，以研究 TGF-β 家族信号如何控制乳腺癌细胞的浸润、渗出，并调节肿瘤血管生成。将荧光标记的 mCherry 乳腺癌细胞注射到 Tg（fli：EGFP）转基因 Casper 斑马鱼胚胎的卵黄囊周围或杜氏管中，在这些斑马鱼中，整个脉管系统都表达增强型绿色荧光蛋白。使用荧光显微镜观察迁移和侵袭性人类癌细胞的动态反应，以及癌细胞在斑马鱼宿主中诱导新血管形成的情况。这些实验提供了高效、可靠、低成本的模型，可用于研究（表观遗传）遗传调节剂和药理学化合物对 TGF-β 家族信号成分活性的干扰对乳腺癌细胞转移和血管生成的影响。
