Embryonic zebrafish xenograft assay of human cancer metastasis

Cancer metastasis is the most important prognostic factor determining patient survival, but currently there are very few drugs or therapies that specifically inhibit the invasion and metastasis of cancer cells. Currently, human cancer metastasis is largely studied using transgenic and immunocompromised mouse xenograft models, which are useful for analysing end-point tumour growth but are unable to accurately and reliably monitor in vivo invasion, intravasation, extravasation or secondary tumour formation of human cancer cells. Furthermore, limits in our ability to accurately monitor early stages of tumour growth and detect micro-metastases likely results in pain and suffering to the mice used for cancer xenograft experiments. Zebrafish (Danio rerio) embryos, however, offer many advantages as a model system for studying the complex, multi-step processes involved during cancer metastasis. This article describes a detailed method for the analysis of human cancer cell invasion and metastasis in zebrafish embryos before they reach protected status at 5 days post fertilisation. Results demonstrate that human cancer cells actively invade within a zebrafish microenvironment, and form metastatic tumours at secondary tissue sites, suggesting that the mechanisms involved during the different stages of metastasis are conserved between humans and zebrafish, supporting the use of zebrafish embryos as a viable model of human cancer metastasis. We suggest that the embryonic zebrafish xenograft model of human cancer is a tractable laboratory model that can be used to understand cancer biology, and as a direct replacement of mice for the analysis of drugs that target cancer invasion and metastasis.

Download Full-text

Abstract 2335: Oncogenic K-RAS activation promotes cell motility, invasion, tumor growth and cancer metastasis via its most downstream signaling module, the SIAH2-dependent proteolytic machinery, in the highly aggressive and metastatic human cancer cells

10.1158/1538-7445.am2011-2335 ◽

2011 ◽

Author(s):

Yang Liao ◽

Rebecca L. Schmidt ◽

Zandra E. Walton ◽

Rie Takahashi ◽

Amy H. Tang

Keyword(s):

Tumor Growth ◽

Cell Motility ◽

Cancer Cells ◽

Cancer Metastasis ◽

Human Cancer ◽

Downstream Signaling ◽

Human Cancer Cells ◽

Ras Activation

Download Full-text

Deficiency of Ku Induces Host Cell Exploitation in Human Cancer Cells

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.651818 ◽

2021 ◽

Vol 9 ◽

Author(s):

Okay Saydam ◽

Nurten Saydam

Keyword(s):

Host Cell ◽

Cancer Cells ◽

Cell Line ◽

Cancer Cell ◽

Cancer Metastasis ◽

Human Cancer ◽

Therapy Resistance ◽

Host Cells ◽

Human Cancer Cells ◽

Parasitic Lifestyle

Cancer metastasis is the major cause of death from cancer (Massague and Obenauf, 2016; Steeg, 2016). The extensive genetic heterogeneity and cellular plasticity of metastatic tumors set a prime barrier for the current cancer treatment protocols (Boumahdi and de Sauvage, 2020). In addition, acquired therapy resistance has become an insurmountable obstacle that abolishes the beneficial effects of numerous anti-cancer regimens (De Angelis et al., 2019; Boumahdi and de Sauvage, 2020). Here we report that deficiency of Ku leads to the exploitation of host cells in human cancer cell line models. We found that, upon conditional deletion of XRCC6 that codes for Ku70, HCT116 human colorectal cancer cells gain a parasitic lifestyle that is characterized by the continuous cycle of host cell exploitation. We also found that DAOY cells, a human medulloblastoma cell line, innately lack nuclear Ku70/Ku86 proteins and utilize the host-cell invasion/exit mechanism for maintenance of their survival, similarly to the Ku70 conditionally-null HCT116 cells. Our study demonstrates that a functional loss of Ku protein promotes an adaptive, opportunistic switch to a parasitic lifestyle in human cancer cells, providing evidence for a previously unknown mechanism of cell survival in response to severe genomic stress. We anticipate that our study will bring a new perspective for understanding the mechanisms of cancer cell evolution, leading to a shift in the current concepts of cancer therapy protocols directed to the prevention of cancer metastasis and therapy resistance.

Download Full-text

Ras Farnesylation Inhibitor FTI-277 Restores the E-Cadherin/Catenin Cell Adhesion System in Human Cancer Cells and Reduces Cancer Metastasis

Japanese Journal of Cancer Research ◽

10.1111/j.1349-7006.2002.tb02479.x ◽

2002 ◽

Vol 93 (9) ◽

pp. 1020-1028 ◽

Cited By ~ 16

Author(s):

Jeong-Seok Nam ◽

Yoshinori Ino ◽

Michiie Sakamoto ◽

Setsuo Hirohashi

Keyword(s):

Cell Adhesion ◽

Cancer Cells ◽

Cancer Metastasis ◽

Human Cancer ◽

Human Cancer Cells ◽

Adhesion System ◽

E Cadherin

Download Full-text

Morphological Abnormalities and Gene Expression Changes Caused by High Incubation Temperatures in Zebrafish Xenografts with Human Cancer Cells

Genes ◽

10.3390/genes12010113 ◽

2021 ◽

Vol 12 (1) ◽

pp. 113

Author(s):

Pablo Cabezas-Sainz ◽

Carlos Coppel ◽

Alba Pensado-López ◽

Pedro Fernandez ◽

Laura Muinelo-Romay ◽

...

Keyword(s):

Cancer Cells ◽

Incubation Temperature ◽

Human Cancer ◽

Effect Of Temperature ◽

Zebrafish Embryos ◽

Morphological Abnormalities ◽

Human Cancer Cells ◽

Developmental Effects ◽

Different Temperatures ◽

Incubation Temperatures

Published studies show that most of the human cancer xenograft studies in zebrafish embryos have used incubation temperatures in the range of 32–34 °C for 3–6 days post-injection, trying to find a compromise temperature between the zebrafish embryos (28 °C) and the human injected cells (37 °C). While this experimental setup is widely used, a question remains: is possible to overcome the drawbacks caused by a suboptimal temperature for the injected cells? To clarify the effect of temperature and injected cells on the host, in this study, we analyzed the development and health of the last in response to different temperatures in the presence or absence of injected human cancer cells. Comparing different incubation temperatures (28, 34 and 36 °C), we determined morphological abnormalities and developmental effects in injected and non-injected embryos at different time points. Besides this, the expression of selected genes was determined by qPCR to determine temperature affected metabolic processes in the embryos. The results indicate that an incubation temperature of 36 °C during a period of 48 h is suitable for xenotransplantation without morphological or metabolic changes that could be affecting the host or the injected cells, allowing them to proliferate near their optimal temperature.

Download Full-text

ZeOncoTest: Refining and Automating the Zebrafish Xenograft Model for Drug Discovery in Cancer

Pharmaceuticals ◽

10.3390/ph13010001 ◽

2019 ◽

Vol 13 (1) ◽

pp. 1 ◽

Cited By ~ 5

Author(s):

Carles Cornet ◽

Sylvia Dyballa ◽

Javier Terriente ◽

Valeria Di Giacomo

Keyword(s):

Drug Discovery ◽

Cancer Cells ◽

Human Cancer ◽

Xenograft Model ◽

Cancer Cell Line ◽

Metastatic Potential ◽

Cell Labeling ◽

Human Cancer Cells ◽

Zebrafish Larvae ◽

Colorectal Cancer Cells

The xenograft of human cancer cells in model animals is a powerful tool for understanding tumor progression and metastatic potential. Mice represent a validated host, but their use is limited by the elevated experimental costs and low throughput. To overcome these restrictions, zebrafish larvae might represent a valuable alternative. Their small size and transparency allow the tracking of transplanted cells. Therefore, tumor growth and early steps of metastasis, which are difficult to evaluate in mice, can be addressed. In spite of its advantages, the use of this model has been hindered by lack of experimental homogeneity and validation. Considering these facts, the aim of our work was to standardize, automate, and validate a zebrafish larvae xenograft assay with increased translatability and higher drug screening throughput. The ZeOncoTest reliability is based on the optimization of different experimental parameters, such as cell labeling, injection site, automated individual sample image acquisition, and analysis. This workflow implementation finally allows a higher precision and experimental throughput increase, when compared to previous reports. The approach was validated with the breast cancer cell line MDA-MB-231, the colorectal cancer cells HCT116, and the prostate cancer cells PC3; and known drugs, respectively RKI-1447, Docetaxel, and Mitoxantrone. The results recapitulate growth and invasion for all tested tumor cells, along with expected efficacy of the compounds. Finally, the methodology has proven useful for understanding specific drugs mode of action. The insights gained bring a step further for zebrafish larvae xenografts to enter the regulated preclinical drug discovery path.

Download Full-text