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Molecular Imaging and Biology

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01-04-2016 | Brief Article

Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine

Authors: Laura L. Bronsart, Christian Stokes, Christopher H. Contag

Published in: Molecular Imaging and Biology | Issue 2/2016

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.

Szatrowski TP, Nathan CF (1991) Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 51:794–798PubMed

Waris G, Ahsan H (2006) Reactive oxygen species: role in the development of cancer and various chronic conditions. J Carcinog 5:14CrossRefPubMedPubMedCentral

Liou G-Y, Storz P (2010) Reactive oxygen species in cancer. Free Radic Res 44:479–496CrossRefPubMed

Bartsch H, Nair J (2004) Oxidative stress and lipid peroxidation-derived DNA-lesions in inflammation driven carcinogenesis. Cancer Detect Prev 28:385–391CrossRefPubMed

Bartsch H, Nair J (2002) Potential role of lipid peroxidation derived DNA damage in human colon carcinogenesis: studies on exocyclic base adducts as stable oxidative stress markers. Cancer Detect Prev 26:308–312CrossRefPubMed

Maynard S, Schurman SH, Harboe C et al (2009) Base excision repair of oxidative DNA damage and association with cancer and aging. Carcinogenesis 30:2–10CrossRefPubMedPubMedCentral

Wiseman H, Halliwell B (1996) Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. Biochem J 313(Pt 1):17–29CrossRefPubMedPubMedCentral

Burhans WC, Heintz NH (2009) The cell cycle is a redox cycle: linking phase-specific targets to cell fate. Free Radic Biol Med 47:1282–1293CrossRefPubMed

Dickinson BC, Chang CJ (2011) Chemistry and biology of reactive oxygen species in signaling or stress responses. Nat Chem Biol 7:504–511CrossRefPubMedPubMedCentral

10.

Hensley K, Robinson KA, Gabbita SP et al (2000) Reactive oxygen species, cell signaling, and cell injury. Free Radic Biol Med 28:1456–1462CrossRefPubMed

11.

Ray PD, Huang BW, Tsuji Y (2012) Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cell Signal 24:981–990CrossRefPubMedPubMedCentral

12.

Arnold RS, Shi J, Murad E et al (2001) Hydrogen peroxide mediates the cell growth and transformation caused by the mitogenic oxidase Nox1. Proc Natl Acad Sci U S A 98:5550–5555CrossRefPubMedPubMedCentral

13.

Irani K, Xia Y, Zweier JL et al (1997) Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts. Science 275:1649–1652CrossRefPubMed

14.

Wang X, Martindale JL, Liu Y, Holbrook NJ (1998) The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 333(Pt 2):291–300CrossRefPubMedPubMedCentral

15.

Burdon RH (1995) Superoxide and hydrogen peroxide in relation to mammalian cell proliferation. Free Radic Biol Med 18:775–794CrossRefPubMed

16.

Cai H, Dikalov S, Griendling KK, Harrison DG (2007) Detection of reactive oxygen species and nitric oxide in vascular cells and tissues: comparison of sensitivity and specificity. Methods Mol Med 139:293–311CrossRefPubMed

17.

Zhao H, Kalivendi S, Zhang H et al (2003) Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. Free Radic Biol Med 34:1359–1368CrossRefPubMed

18.

Nazarewicz RR, Bikineyeva A, Dikalov SI (2013) Rapid and specific measurements of superoxide using fluorescence spectroscopy. J Biomol Screen 18:498–503CrossRefPubMedPubMedCentral

19.

Teranishi K, Shimomura O (1997) Coelenterazine analogs as chemiluminescent probe for superoxide anion. Anal Biochem 249:37–43CrossRefPubMed

20.

Tarpey MM, White CR, Suarez E et al (1999) Chemiluminescent detection of oxidants in vascular tissue. Lucigenin but not coelenterazine enhances superoxide formation. Circ Res 84:1203–1211CrossRefPubMed

21.

Lucas M, Solano F (1992) Coelenterazine is a superoxide anion-sensitive chemiluminescent probe: its usefulness in the assay of respiratory burst in neutrophils. Anal Biochem 206:273–277CrossRefPubMed

22.

Wu C, Nakamura H, Murai A, Shimomura O (2001) Chemi- and bioluminescence of coelenterazine analogues with a conjugated group at the C-8 position. Tetrahedron Lett 42:2997–3000CrossRef

23.

Chen SF, Navizet I, Roca-Sanjuán D et al (2012) Chemiluminescence of coelenterazine and fluorescence of coelenteramide: a systematic theoretical study. J Chem Theory Comput 8:2796–2807CrossRefPubMed

24.

Fernandes DC, Wosniak J, Pescatore LA et al (2007) Analysis of DHE-derived oxidation products by HPLC in the assessment of superoxide production and NADPH oxidase activity in vascular systems. Am J Physiol Cell Physiol 292:C413–C422CrossRefPubMed

25.

Zhao H, Doyle TC, Wong RJ et al (2004) Characterization of coelenterazine analogs for measurements of Renilla luciferase activity in live cells and living animals. Mol Imaging 3:43–54CrossRefPubMed

26.

Mukhopadhyay P, Rajesh M, Yoshihiro K et al (2007) Simple quantitative detection of mitochondrial superoxide production in live cells. Biochem Biophys Res Commun 358:203–208CrossRefPubMedPubMedCentral

27.

Caldefie-Chézet F, Walrand S, Moinard C et al (2002) Is the neutrophil reactive oxygen species production measured by luminol and lucigenin chemiluminescence intra or extracellular? Comparison with DCFH-DA flow cytometry and cytochrome c reduction. Clin Chim Acta 319:9–17CrossRefPubMed

Title: Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine
Authors: Laura L. Bronsart
Christian Stokes
Christopher H. Contag
Publication date: 01-04-2016
Publisher: Springer US
Published in: Molecular Imaging and Biology / Issue 2/2016
Print ISSN: 1536-1632
Electronic ISSN: 1860-2002
DOI: https://doi.org/10.1007/s11307-015-0896-7

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine

Abstract

Purpose

Procedures

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Procedures

Results

Conclusions

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