Luciferase expression and bioluminescence does not affect tumor cell growth in vitro or in vivo

Molecular Cancer

Short Communication

Cynthia Ng¹  Jessamy C Tiffen¹  Jeff Holst¹  Charles G Bailey¹  John EJ Rasko² 

[1] Gene and Stem Cell Therapy Program, Centenary Institute, University of Sydney, 2050, Camperdown, NSW, Australia;Gene and Stem Cell Therapy Program, Centenary Institute, University of Sydney, 2050, Camperdown, NSW, Australia;Cell and Molecular Therapies, Royal Prince Alfred Hospital, 2050, Camperdown, NSW, Australia;

关键词: Green Fluorescent Protein;    Green Fluorescent Protein Expression;    Luciferase Expression;    Bioluminescence Imaging;    Biophotonic Emission;   

DOI  :  10.1186/1476-4598-9-299

 received in 2010-05-12, accepted in 2010-11-22,  发布年份 2010

来源: Springer

【 摘 要 】

Live animal imaging is becoming an increasingly common technique for accurate and quantitative assessment of tumor burden over time. Bioluminescence imaging systems rely on a bioluminescent signal from tumor cells, typically generated from expression of the firefly luciferase gene. However, previous reports have suggested that either a high level of luciferase or the resultant light reaction produced upon addition of D-luciferin substrate can have a negative influence on tumor cell growth. To address this issue, we designed an expression vector that allows simultaneous fluorescence and luminescence imaging. Using fluorescence activated cell sorting (FACS), we generated clonal cell populations from a human breast cancer (MCF-7) and a mouse melanoma (B16-F10) cell line that stably expressed different levels of luciferase. We then compared the growth capabilities of these clones in vitro by MTT proliferation assay and in vivo by bioluminescence imaging of tumor growth in live mice. Surprisingly, we found that neither the amount of luciferase nor biophotonic activity was sufficient to inhibit tumor cell growth, in vitro or in vivo. These results suggest that luciferase toxicity is not a necessary consideration when designing bioluminescence experiments, and therefore our approach can be used to rapidly generate high levels of luciferase expression for sensitive imaging experiments.

【 授权许可】

Unknown   
© Tiffen et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.