Abstract
Respiration is one of the major functions of mitochondria, whereby these vital organelles use oxygen to produce energy. Many agents that may be of potential clinical relevance act by targeting mitochondria, where they may suppress mitochondrial respiration. It is therefore important to evaluate this process and understand how this is modulated by small molecules. Here, we describe the general methodology to assess respiration in cultured cells, followed by the evaluation of the effect of one anticancer agent targeted to mitochondria on this process, and also how to assess this in tumor tissue.
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References
Galluzzi L, Kepp O, Trojel-Hansen C, Kroemer G (2012) Mitochondrial control of cellular life, stress, and death. Circ Res 111:1198–1207
Murphy MP (2009) How mitochondria produce reactive oxygen species. Biochem J 417:1–13
Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674
Trachootham D, Alexandre J, Huang P (2009) Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach? Nat Rev Drug Discov 8:579–591
Vander Heiden MG, Cantley LC, Thompson CB (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324:1029–1033
Fulda S, Galluzzi L, Kroemer G (2010) Targeting mitochondria for cancer therapy. Nat Rev Drug Discov 9:447–464
Neuzil J, Dong LF, Rohlena J, Truksa J, Ralph SJ (2013) Classification of mitocans, anti-cancer drugs acting on mitochondria. Mitochondrion 13:199–208
Rohlena J, Dong LF, Ralph SJ, Neuzil J (2011) Anticancer drugs targeting the mitochondrial electron transport chain. Antioxid Redox Signal 15:2951–2974
Pesta D, Gnaiger E (2012) High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol 810:25–58
Kuznetsov AV, Veksler V, Gellerich FN, Saks V, Margreiter R, Kunz WS (2008) Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nat Protoc 3:965–976
Dong LF, Jameson VJ, Tilly D, Cerny J, Mahdavian E, Marin-Hernandez A, Hernandez-Esquivel L, Rodriguez-Enriquez S, Stursa J, Witting PK, Stantic B, Rohlena J, Truksa J, Kluckova K, Dyason JC, Ledvina M, Salvatore BA, Moreno-Sanchez R, Coster MJ, Ralph SJ, Smith RA, Neuzil J (2011) Mitochondrial targeting of vitamin E succinate enhances its pro-apoptotic and anti-cancer activity via mitochondrial complex II. J Biol Chem 286:3717–3728
Dong LF, Jameson VJ, Tilly D, Prochazka L, Rohlena J, Valis K, Truksa J, Zobalova R, Mahdavian E, Kluckova K, Stantic M, Stursa J, Freeman R, Witting PK, Norberg E, Goodwin J, Salvatore BA, Novotna J, Turanek J, Ledvina M, Hozak P, Zhivotovsky B, Coster MJ, Ralph SJ, Smith RA, Neuzil J (2011) Mitochondrial targeting of α-tocopheryl succinate enhances its pro-apoptotic efficacy: a new paradigm for effective cancer therapy. Free Radic Biol Med 50:1546–1555
Rohlena J, Dong LF, Kluckova K, Zobalova R, Goodwin J, Tilly D, Stursa J, Pecinova A, Philimonenko A, Hozak P, Banerjee J, Ledvina M, Sen CK, Houstek J, Coster MJ, Neuzil J (2011) Mitochondrially targeted α-tocopheryl succinate is antiangiogenic: potential benefit against tumor angiogenesis but caution against wound healing. Antioxid Redox Signal 15:2923–2935
Hroudova J, Fisar Z (2012) In vitro inhibition of mitochondrial respiratory rate by antidepressants. Toxicol Lett 213:345–352
Kovarova J, Bajzikova M, Vondrusova M, Stursa J, Goodwin J, Nguyen M, Zobalova R, Pesdar EA, Truksa J, Tomasetti M, Dong LF, Neuzil J (2014) Mitochondrial targeting of α-tocopheryl succinate enhances its anti-mesothelioma efficacy. Redox Rep 19:16–25
Acknowledgements
This work was supported in part by grants from the National and Health Medical Research Council and the Australian Research Council, and by BIOCEV European Regional Development Fund CZ.1.05/1.1.00/02.0109 and Czech Science Foundation grant no. P301-12-1851.
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Kluckova, K., Dong, LF., Bajzikova, M., Rohlena, J., Neuzil, J. (2015). Evaluation of Respiration of Mitochondria in Cancer Cells Exposed to Mitochondria-Targeted Agents. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1265. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2288-8_14
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DOI: https://doi.org/10.1007/978-1-4939-2288-8_14
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2287-1
Online ISBN: 978-1-4939-2288-8
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