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Resveratrol mobilizes Ca2+ from intracellular stores and induces c-Jun N-terminal kinase activation in tumoral AR42J cells

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Abstract

Resveratrol (3,4′,5-trihydroxy-trans-stilbene), a phytoalexin naturally found in grapes and red wine, is a redox-active compound endowed with significant positive activities. In this study, the effects of resveratrol on intracellular free Ca2+ concentration ([Ca2+]c) and on cell viability in tumoral AR42J pancreatic cells are examined. The results show that resveratrol (100 μM and 1 mM) induced changes in [Ca2+]c, that consisted of single or short lasting spikes followed by a slow reduction toward a value close to the resting level. Lower concentrations of resveratrol (1 and 10 μM) did not show detectable effects on [Ca2+]c. Depletion of intracellular Ca2+ stores by stimulation of cells with 1 nM CCK-8, 20 pM CCK-8 or 1 μM thapsigargin, blocked Ca2+ responses evoked by resveratrol. Conversely, prior stimulation of cells with resveratrol inhibited Ca2+ mobilization in response to a secondary application of CCK-8 or thapsigargin. In addition, resveratrol inhibited oscillations in [Ca2+]c evoked by a physiological concentration of CCK-8 (20 pM). On the other hand, incubation of cells in the presence of resveratrol induced a reduction of cell viability. Finally, incubation of AR42J cells in the presence of resveratrol led to activation of c-Jun N-terminal kinase (JNK), a mitogen-activated protein kinase responsive to stress stimuli. Activation of JNK was reduced in the absence of extracellular Ca2+. In summary, the results show that resveratrol releases Ca2+ from intracellular stores, most probably from the endoplasmic reticulum, and reduces AR42J cells viability. Reorganization of cell’s survival/death processes in the presence of resveratrol may involve Ca2+-mediated JNK activation.

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Abbreviations

[Ca2+]c :

Cytosolic free Ca2+ concentration

CCK-8:

Cholecystokinin octapeptide

EGTA:

Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid

ER:

Endoplasmic reticulum

Fura-2/AM:

Fura-2 acetoxymethyl ester

IP3 :

Inositol 1,4,5-trisphosphate

JNK:

c-Jun N-terminal kinase

PMCA:

Plasma membrane calcium ATPase

ROS:

Reactive oxygen species

SERCA:

Sarcoendoplasmic reticulum Ca2+-ATPase

Tps:

Thapsigargin

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Acknowledgments

The authors declare that there is no conflict of interest. This study was supported by Junta de Extremadura-FEDER (PRI08A018 and GR10010). Patricia Santofimia-Castaño was granted a fellowship from Junta de Extremadura (Consejería de Economía, Comercio e Innovación) and European Social Fund. Alvaro Miro-Moran was granted a fellowship from MICINN-FEDER (BES-2008-002106). The authors would like to thank Mrs. Mercedes Gomez Blazquez for her excellent technical support.

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  1. Cell Physiology Research Group, Department of Physiology, University of Extremadura, Avenida Universidad s/n, 10003, Caceres, Spain

    Lourdes Garcia-Sanchez, Patricia Santofimia-Castaño, Alvaro Miro-Moran, Jose A. Tapia, Gines M. Salido & Antonio Gonzalez

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  1. Lourdes Garcia-Sanchez
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  2. Patricia Santofimia-Castaño
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  3. Alvaro Miro-Moran
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  4. Jose A. Tapia
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  5. Gines M. Salido
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  6. Antonio Gonzalez
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Correspondence to Antonio Gonzalez.

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Garcia-Sanchez, L., Santofimia-Castaño, P., Miro-Moran, A. et al. Resveratrol mobilizes Ca2+ from intracellular stores and induces c-Jun N-terminal kinase activation in tumoral AR42J cells. Mol Cell Biochem 362, 15–23 (2012). https://doi.org/10.1007/s11010-011-1123-8

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