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The Ca2+ channel blocker flunarizine induces caspase-10-dependent apoptosis in Jurkat T-leukemia cells

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Abstract

Flunarizine is a Ca2+ channel blocker that can be either cytoprotective or cytotoxic, depending on the cell type that is being examined. We show here that flunarizine was cytotoxic for Jurkat T-leukemia cells, as well as for other hematological maligancies, but not for breast or colon carcinoma cells. Treatment of Jurkat cells with flunarizine resulted in caspase-3 activation, poly (ADP-ribose) polymerase cleavage, and laddering of DNA fragments, all of which are hallmarks of apoptosis. Flunarizine-induced DNA fragmentation was inhibited by the caspase-3 inhibitor z-DEVD-fmk, the caspase-8/caspase-10 inhibitor z-IETD-fmk, and the caspase-10 inhibitor z-AEVD-fmk, but was not reduced in caspase-8-deficient Jurkat cells, indicating the involvement of caspase-10 upstream of caspase-3 activation. Interestingly, FADD recruitment to a death receptor was not involved since flunarizine caused DNA fragmentation in FADD-deficient Jurkat cells. Flunarizine treatment of Jurkat cells also resulted in reactive oxygen species production, dissipation of mitochondrial transmembrane potential, release of cytochrome c from mitochondria, and caspase-9 activation, although none of these events were necessary for apoptosis induction. Collectively, these findings indicate that flunarizine triggers apoptosis in Jurkat cells via FADD-independent activation of caspase-10. Flunarizine warrants further investigation as a potential anti-cancer agent for the treatment of hematological malignancies.

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Acknowledgments

The authors thank Julie Long for technical assistance. This research was supported by a grant to D.H. from the Natural Sciences and Engineering Research Council (NSERC). D.C. was the recipient of postgraduate scholarships from NSERC and the Nova Scotia Health Research Foundation. S.F. and C.D. are recipients of NSERC postgraduate scholarships.

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Authors and Affiliations

  1. Department of Microbiology & Immunology, Dalhousie University, Halifax, Nova Scotia, B3H 1X5, Canada

    David M. Conrad, Suzanne J. Furlong, Kenneth A. West & David W. Hoskin

  2. Department of Pathology, Dalhousie University, Halifax, Nova Scotia, B3H 1X5, Canada

    Carolyn D. Doucette, Kenneth A. West & David W. Hoskin

  3. Department of Medicine, Dalhousie University, Halifax, Nova Scotia, B3H 1X5, Canada

    Kenneth A. West

  4. Department of Surgery, Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia, B3H 1X5, Canada

    David W. Hoskin

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  1. David M. Conrad
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Correspondence to David W. Hoskin.

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David M. Conrad and Suzanne J. Furlong contributed equally to this work.

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Conrad, D.M., Furlong, S.J., Doucette, C.D. et al. The Ca2+ channel blocker flunarizine induces caspase-10-dependent apoptosis in Jurkat T-leukemia cells. Apoptosis 15, 597–607 (2010). https://doi.org/10.1007/s10495-010-0454-3

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