Abstract
Purpose
We evaluated the small molecule coelenterazine as a potential reporter of cancer-associated superoxide anion in cell culture and in mice.
Procedures
The superoxide anion concentrations of various cancer cell lines were quantified by coelenterazine chemiluminescence in vitro. Coelenteramide fluorescence was detected via flow cytometry and fluorescent microscopy. Coelenterazine was used for the in vivo detection of cancer-associated superoxide anion using the 4T1 breast adenocarcinoma mouse model.
Results
Various cell lines in culture demonstrated different superoxide anion concentrations, with a signal range of 3.15 ± 0.06 to 11.80 ± 0.24 times that of background. In addition to chemiluminescent detection of coelenterazine, we demonstrated fluorescent detection of coelenteramide within the cytoplasm of cells. 4T1 murine mammary adenocarcinoma tumors in mice demonstrated significantly higher 2.13 ± 0.19-fold coelenterazine-based chemiluminescence than that of surrounding normal tissues.
Conclusions
Collectively, our results indicate that coelenterazine can be used to assay superoxide anion concentrations in cultured cancer cells and in tumors growing in mice.
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Acknowledgments
This work was generously supported, in part, by an unrestricted gift from the Chamber’s Family Foundation (to C.H.C.) and by a grant from the National Institutes of Health (1R24DK096465-01).
Conflict of Interest
The authors declare that they have no competing interests.
Ethics Statement
All applicable institutional and/or national guidelines for the care and use of animals were followed.
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Bronsart, L.L., Stokes, C. & Contag, C.H. Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine. Mol Imaging Biol 18, 166–171 (2016). https://doi.org/10.1007/s11307-015-0896-7
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DOI: https://doi.org/10.1007/s11307-015-0896-7