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Garlic compounds induced calpain and intrinsic caspase cascade for apoptosis in human malignant neuroblastoma SH-SY5Y cells

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Abstract

Malignant (N-type) neuroblastoma continues to defy current chemotherapeutic regimens. We tested the garlic compounds diallyl sulfide (DAS) and diallyl disulfide (DADS) for induction of apoptosis in human malignant neuroblastoma SH-SY5Y cells. Viability of human primary neurons was unaffected after 24 h treatment with 50 and 100 μM DAS and 50 μM DADS but slightly affected with 100 μM DADS. Treatment with 50 and 100 μM DAS or DADS significantly decreased viability in SH-SY5Y cells. Wright staining showed morphological features of apoptosis in SH-SY5Y cells treated with 50 and 100 μM DAS or DADS for 24 h. ApopTag assay demonstrated DNA fragmentation in apoptotic cells. Apoptosis was associated with an increase in [Ca2+]i, increase in Bax:Bcl-2 ratio, mitochondrial release of cytochrome c, increase in cytosolic Smac/Diablo, and down regulation of inhibitor-of-apoptosis proteins and nuclear factor kappa B (NFκB). Activation of caspase-9 and caspase-3 indicated involvement of intrinsic pathway of apoptosis. Calpain and caspase-3 activities produced 145 kD spectrin break down product (SBDP) and 120 kD SBDP, respectively. Also, caspase-3 activity cleaved inhibitor of caspase-activated DNase (ICAD). Results strongly suggested that the garlic compounds DAS and DADS suppressed anti-apoptotic factors and activated calpain and intrinsic caspase cascade for apoptosis in SH-SY5Y cells.

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Abbreviations

BIRC-2::

Baculovirus inhibitor-of-apoptosis repeat containing-2

BIRC-3::

Baculovirus inhibitor-of-apoptosis repeat containing-3

[Ca2+]i::

Intracellular free calcium concentration

CAD::

Caspase-activated DNase

c-IAP1::

Cellular inhibitor-of-apoptosis protein 1

c-IAP2::

Cellular inhibitor-of-apoptosis protein 2

DADS::

Diallyl disulfide

DAS::

Diallyl sulfide

IAPs::

Inhibitor-of-apoptosis proteins

ICAD::

Inhibitor of caspase-activated DNase

IκВα::

Inhibitor of nuclear factor kappa B alpha

NFκВ::

Nuclear factor kappa B

SBDP::

Spectrin breakdown product

Smac/Diablo::

Second mitochondrial activator of caspases/direct IAP binding protein with low pI

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Acknowledgements

This investigation was supported in part by the R01 grants (CA-91460 and NS-57811) from the National Institutes of Health (Bethesda, MD, USA) and also by the Spinal Cord Injury Research Fund (SCIRF-0803) from the State of South Carolina.

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  1. Department of Neurosciences, Medical University of South Carolina (MUSC), 96 Jonathan Lucas Street, Suite 323K, P.O. Box 250606, Charleston, SC, 29425, USA

    Surajit Karmakar, Naren L. Banik, Sunil J. Patel & Swapan K. Ray

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  1. Surajit Karmakar
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  2. Naren L. Banik
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Correspondence to Swapan K. Ray.

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Karmakar, S., Banik, N.L., Patel, S.J. et al. Garlic compounds induced calpain and intrinsic caspase cascade for apoptosis in human malignant neuroblastoma SH-SY5Y cells. Apoptosis 12, 671–684 (2007). https://doi.org/10.1007/s10495-006-0024-x

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