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Trifolin acetate-induced cell death in human leukemia cells is dependent on caspase-6 and activates the MAPK pathway

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Abstract

In the present study we demonstrated that the flavonoid derivative trifolin acetate (TA), obtained by acetylation of naturally occurring trifolin, induces apoptosis. Associated downstream signaling events were also investigated. TA-induced cell death was prevented by the non-specific caspase inhibitor z-VAD-fmk and reduced by the presence of the selective caspase inhibitors z-LEHD-fmk (caspase-9), z-DEVD-fmk (caspase-3) and z-VEID-fmk (caspase-6). The apoptotic effect of TA was associated with (i) the release of cytochrome c from mitochondria which was not accompanied by dissipation of the mitochondrial membrane potential (ΔΨm), (ii) the activation of the mitogen-activated protein kinases (MAPKs) pathway and (iii) abrogated by the over-expression of Bcl-2 or Bcl-xL. TA-induced cell death was attenuated by inhibition of extracellular signal-regulated kinases (ERK) 1/2 with U0126 and inhibition of p38MAPK with SB203580. In contrast, inhibition of c-Jun NH2-terminal kinase (JNK) by SP600125 significantly enhanced apoptosis. Although reactive oxygen species (ROS) increased in response to TA, this did not seem to play a pivotal role in the apoptotic process since different anti-oxidants were unable to provide cell protection. The present study demonstrates that TA-induced cell death is mediated by an intrinsic-dependent apoptotic event involving mitochondria and MAPK, and through a mechanism independent of ROS generation.

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Acknowledgments

We thank Dr. Angelika Vollmar and Dr. Jacqueline Bréard for supplying HL-60/neo, HL-60/Bcl-xL and U937/Bcl-2 cells, respectively. We thank J. Estévez (Hospital Universitario Insular de Gran Canaria) for his collaboration in the Western blot assays. This work was supported by a Grant from the Ministry of Education and Science of Spain and from the European Regional Development Fund (SAF2004-07928) to FE. FT was supported by a research studentship from the Canary Islands Government.

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  1. Department of Biochemistry, Instituto Canario de Investigación del Cáncer (I.C.I.C.), University of Las Palmas de Gran Canaria, Plaza Dr. Pasteur s/n, 35016, Las Palmas de Gran Canaria, Spain

    Fernando Torres, José Quintana & Francisco Estévez

  2. Department of Chemistry, University of La Laguna, Instituto Universitario de Bio-Orgánica “Antonio González”, 38206, La Laguna, Teneriffe, Spain

    Jesús G. Díaz & Armando J. Carmona

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  1. Fernando Torres
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  2. José Quintana
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  3. Jesús G. Díaz
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  5. Francisco Estévez
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Correspondence to Francisco Estévez.

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Torres, F., Quintana, J., Díaz, J.G. et al. Trifolin acetate-induced cell death in human leukemia cells is dependent on caspase-6 and activates the MAPK pathway. Apoptosis 13, 716–728 (2008). https://doi.org/10.1007/s10495-008-0202-0

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