Invasive Behavior of Human Breast Cancer Cells in Embryonic Zebrafish

Abstract

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.

Deubiquitinase Activity Profiling Identifies UCHL1 as a Candidate Oncoprotein That Promotes TGFβ-Induced Breast Cancer Metastasis

“…Transgenic zebrafish lines Tg (fli1: EGFP) were raised according to standard procedures in compliance with the local Institutional Committee for Animal Welfare of the Leiden University. Zebrafish extravasation assays were prepared as previously described (21). Zebrafish were fixed with 4% paraformaldehyde 6 days after injection.…”

A Small-Molecule Activity-Based Probe for Monitoring Ubiquitin C-terminal Hydrolase L1 (UCHL1) Activity in Live Cells and Zebrafish Embryos

“…To investigate the application of 8RK59 in tracking UCHL1 activity in an in vivo model, we chose the zebrafish ( Danio rerio ) due to their high genetic homology to humans and the transparency of their embryos. 39 Firstly, we treated zebrafish embryos with 8RK59 and recorded fluorescent images during the development of embryos from 1 to 7 days post fertilization (dpf). Results showed 8RK59 mainly labeled the nose, eye and brain of the zebrafish embryos (Figure 6A).…”

Dasatinib inhibition of cSRC prevents the migration and metastasis of canine mammary cancer cells with enhanced Wnt and HER signalling

“…CMT‐U27 RED cells were injected in the yolk sac or in the duct of Cuvier . Cells injected into the yolk sac have to migrate to the blood circulation before they may form distant metastases, whereas direct injection into the bloodstream using the duct of Cuvier does not require intravasation and can form distant metastases more easily …”

“…50 Cells injected into the yolk sac have to migrate to the blood circulation before they may form distant metastases, whereas direct injection into the bloodstream using the duct of Cuvier does not require intravasation and can form distant metastases more easily. 51,52 The embryos were directly inspected for proper injection and were fixed on day 3 or 5 (dpi) and imaged under a confocal microscope. The experiments were performed in triplicate with 30 to40 well-injected embryos for dasatinib and 10 embryos for the everolimus experiments.…”