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Camalexin induces apoptosis in T-leukemia Jurkat cells by increased concentration of reactive oxygen species and activation of caspase-8 and caspase-9

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Abstract

Camalexin, a major indole phytoalexin of Arabidopsis thaliana, accumulates in various cruciferous plants in response to environmental stress and reportedly displays antimicrobial activities against various plant pathogens. However, its cytotoxicity against eukaryotic cells and potential as a prospective drug for human diseases has been examined only in a limited context. Our data demonstrate the time- and concentration-dependent cytotoxicity of camalexin on human T-leukemia Jurkat cells in the micromolar range, and the lower potency of cytotoxic effects on human lymphoblasts and primary fibroblasts. Cytotoxicity of camalexin is enhanced by the glutathione-depleting agent buthionine sulfoximine and completely blocked by pan-caspase inhibitor Z-VAD-FMK. Treatment of Jurkat cells with camalexin resulted in activation of caspase-8, caspase-9, caspases-3/7, and apoptosis that was detected by the presence of a sub-G1 population of cells, externalization of phosphatidyl serine and decreased mitochondrial membrane potential. Staining with 2′,7′-dichlorodihydrofluorescein diacetate and dihydroethidium bromide displayed increased concentration of reactive oxygen species (ROS) early in camalexin-treated Jurkat cells, prior to the onset of apoptosis, while staining with MitoSOX™ dye identified mitochondria as a source of increased ROS. Our data suggest that this phytochemical, which has a wide range of predicted pharmacological activities, induces apoptosis in Jurkat leukemia cells through increased ROS followed by dissipation of mitochondrial membrane potential and execution of caspase-9- and caspase-8-initiated apoptosis. This is, to the best of our knowledge, the first report on antileukemic activity and mode of action of this unique indole phytoalexin.

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Acknowledgments

This work was supported by Deborah Nash Harris Endowment Fund; Ovarian Cycle Foundation, the Ovarian Cancer Institute, and the Slovak Research and Development Agency under the contract No. APVV-0514-06. The authors thank Dr. DeEtte Walker for reviewing the manuscript and Dr. Kenneth Scarberry for his help with the artwork.

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The authors declare that no actual or potential conflict of interest exists in relation to this article.

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Authors and Affiliations

  1. School of Biology, Georgia Institute of Technology, 310 Ferst Dr, Atlanta, GA, 30332, USA

    Roman Mezencev, Taylor Updegrove & John F. McDonald

  2. Faculty of Science, Institute of Chemical Sciences, P. J. Šafárik University, Moyzesova 11, 040 01, Kosice, Slovakia

    Peter Kutschy & Mária Repovská

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  1. Roman Mezencev
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  2. Taylor Updegrove
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Correspondence to Roman Mezencev.

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Mezencev, R., Updegrove, T., Kutschy, P. et al. Camalexin induces apoptosis in T-leukemia Jurkat cells by increased concentration of reactive oxygen species and activation of caspase-8 and caspase-9. J Nat Med 65, 488–499 (2011). https://doi.org/10.1007/s11418-011-0526-x

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