Cancers 2011, 3(4), 4102-4113; doi:10.3390/cancers3044102

Discriminating Different Cancer Cells Using a Zebrafish in Vivo Assay

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Received: 8 September 2011; in revised form: 13 October 2011 / Accepted: 21 October 2011 / Published: 31 October 2011
(This article belongs to the Special Issue Tumor Stroma)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: Despite the expanded understanding of tumor angiogenesis phenomenon and how it impacts cancer treatment outcomes, we have yet to develop a robust assay that can quickly, easily, and quantitatively measure tumor-induced angiogenesis. Since the zebrafish/tumor xenograft represents an emerging tool in this regard, the present study strives to capitalize on the ease, effectiveness, and the adaptability of this model to quantify tumor angiogenesis. In order to test a range of responses, we chose two different tumorigenic cell lines, the human non-small cell lung carcinoma (H1299) and the mouse lung adenocarcinoma (CL13). Non-tumorigenic 3T3-L1 cells served as negative control. The cells were grafted near to the perivitelline space of the zebrafish embryos and the angiogenic response was analyzed using whole-mount alkaline phosphatase (AP) vessel staining and fluorescence microscopy. Angiogenic activity was scored based on the length and number of the newly formed ectopic vessels and the percentage of embryos with ectopic vessels. At 2 day-post-implantation, we detected a significant increase in the length and number of ectopic vessels with H1299 cell implantation compared to CL13 cell transplantation, both are higher than 3T3-L1 control. We also observed a significantly higher percentage of embryos with ectopic vessels with H1299 and CL13 transplantation compared to the 3T3-L1 control, but this parameter is not as robust and reliable as measuring the length and number of ectopic vessels. Furthermore, the systemic exposure of zebrafish embryos to an anti-angiogenesis drug (PTK 787, inhibitor of vascular endothelial growth factor receptor tyrosine kinase) inhibited tumor-induced angiogenesis, suggesting that the assay can be used to evaluate anti-angiogenic drugs. This study implicates the feasibility of using zebrafish xenotransplantation to perform quantitative measurement of the angiogenic activity of cancer cells which can be further extended to measure cancer cell metastasis. This assay represents not only the useful test for patient diagnosis, but also has the potential for evaluating anti-cancer drugs treatment.
Keywords: tumor xenograft; angiogenesis; H1299; CL13; lung carcinoma; cancer cells; zebrafish; subintestinal vessel plexus; PTK787; anti-angiogenesis drug
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MDPI and ACS Style

Moshal, K.S.; Ferri-Lagneau, K.F.; Haider, J.; Pardhanani, P.; Leung, T. Discriminating Different Cancer Cells Using a Zebrafish in Vivo Assay. Cancers 2011, 3, 4102-4113.

AMA Style

Moshal KS, Ferri-Lagneau KF, Haider J, Pardhanani P, Leung T. Discriminating Different Cancer Cells Using a Zebrafish in Vivo Assay. Cancers. 2011; 3(4):4102-4113.

Chicago/Turabian Style

Moshal, Karni S.; Ferri-Lagneau, Karine F.; Haider, Jamil; Pardhanani, Pooja; Leung, TinChung. 2011. "Discriminating Different Cancer Cells Using a Zebrafish in Vivo Assay." Cancers 3, no. 4: 4102-4113.

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