Abstract
Resveratrol, a natural polyphenolic antioxidant, has been reported to possess the cancer chemopreventive potential in wide range by means of triggering tumor cells apoptosis through various pathways. It induced apoptosis through the activation of the mitochondrial pathway in some kinds of cells. In the present reports, we showed that resveratrol-induced HepG2 cell apoptosis and mitochondrial dysfunction was dependent on the induction of the mitochondrial permeability transition (MPT), because resveratrol caused the collapse of the mitochondrial membrane potential (ΔΨm) with the concomitant release of cytochrome c (Cyt.c). In addition, resveratrol induced a rapid and sustained elevation of intracellular [Ca2+], which compromised the mitochondrial ΔΨm and triggered the process of HepG2 cell apoptosis. In permeabilized HepG2 cells, we further demonstrated that the effect of the resveratrol was indeed synergistic with that of Ca2+ and Ca2+ is necessary for resveratrol-induced MPT opening. Calcium-induced calcium release from mitochondria (mCICR) played a key role in mitochondrial dysfunction and cell apoptosis: (1) mCICR inhibitor, ruthenium red (RR), prevent MPT opening and Cyt.c release; and (2) RR attenuated resveratrol-induced HepG2 cell apoptotic death. Furthermore, resveratrol promotes MPT opening by lowering Ca2+-threshold. These data suggest modifying mCICR and Ca2+ threshold to modulate MPT opening may be a potential target to control cell apoptosis induced by resveratrol.
Similar content being viewed by others
References
Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, Fong HH, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM (1997) Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science 275(5297):218–220
Aziz MH, Kumar R, Ahmad N (2003) Cancer chemoprevention by resveratrol: in vivo and in vivo studies and the underlying mechanisms. Int J Oncol 23(1):17–28
Aggarwal BB, Bhardwaj A, Aggarwal RS, Seeram NP, Shishodia S, Takada Y (2004) Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res 24(5A):2783–2840
Park JW, Choi YJ, Suh SI, Baek WK, Suh MH, Jin IN, Min DS, Woo JH, Chang JS, Passaniti A, Lee YH, Kwon TK (2001) Bcl-2 overexpression attenuates resveratrol-induced apoptosis in U937 cells by inhibition of caspase-3 activity. Carcinogenesis 22(10):1633–1639
Clement MV, Hirpara JL, Chawdhury SH, Pervaiz S (1998) Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling-dependent apoptosis in human tumor cells. Blood 92(3):996–1002
Joe AK, Liu H, Suzui M, Vural ME, Xiao D, Weinstein IB (2002) Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Carcer Res 8(3):893–903
Tinhofer I, Bernhard D, Senfter M, Anether G, Loeffler M, Kroemer G, Kofler R, Csordas A, Greil R (2001) Resveratrol, a tumor-suppressive compound from grapes, induces apoptosis via a novel mitochondrial pathway controlled by Bcl-2. FASEB J 15(9):1613–1615
Mohan J, Gandhi AA, Bhavya BC, Rashmi R, Karunagaran D, Indu R, Santhoshkumar TR (2006) Caspase-2 triggers Bax-Bak-dependent and -independent cell death in colon cancer cells treated with resveratrol. J Biol Chem 281(26):17599–17611
Kim JS, He L, Lemasters JJ (2003) Mitochondrial permeability transition: a common pathway to necrosis and apoptosis. Biochem Biophys Res Commun 304(3):463–470
Green DR, Kroemer G (2004) The pathophysiology of mitochondrial cell death. Science 305(5684):626–629
Gunter TE, Buntinas L, Sparagna G, Eliseev R, Gunter K (2000) Mitochondrial calcium transport: mechanisms and functions. Cell Calcium 28(5–6):285–296
Armstrong JS (2006) Mitochondrial membrane permeabilization: the sine qua non for cell death. Bioessays 28(3):253–260
Halestrap AP, Brennerb C (2003) The adenine nucleotide translocase: a central component of the mitochondrial permeability transition pore and key player in cell death. Curr Med Chem 10(16):1507–1525
Huang X, Zhai D, Huang Y (2001) Dependence of permeability transition pore opening and cytochrome C release from mitochondria on mitochondria energetic status. Mol Cell Biochem 224(1–2):1–7
Smaili SS, Hsu YT, Youle RJ, Russell JT (2000) Mitochondria in Ca2+ signaling and apoptosis. J Bioenerg Biomembr 32(1):35–46
Simpson PB, Russell JT (1998) Role of mitochondrial Ca2+ regulation in neuronal and glial cell signaling. Brain Res Brain Res Rev 26(1):72–81
Ichas F, Jouaville LS, Mazat JP (1997) Mitochondria are excitable organelles capable of generating and conveying electrical and calcium signals. Cell 89(7):1145–1153
Ichas F, Mazat JP (1998) From calcium signaling to cell death: two conformations for the mitochondrial permeability transition pore. Switching from low-to high-conductance state. Biochem Biophys Acta 1366(1–2):33–50
Huang X, Zhai D, Huang Y (2000) Study on the relationship between calcium-induced calcium release from mitochondria and PTP opening. Mol Cell Biochem 213(1–2):29–35
Frambach I, Rossler W, Winkler M, Schurmann FW (2004) F-actin at identified synapses in the mushroom body neuropil of the insect brain. J Comp Neurol 475(3):303–314
Rose P, Whiteman M, Huang SH, Halliwell B, Ong CN (2003) β-Phenylethyl isothiocyanate-mediated apoptosis in hepatoma HepG2 cells. Cell Mol Life Sci 60(7):1489–1503
Oh KW, Qian T, Brenner DA, Lemasters JJ (2003) Salicylate enhances necrosis and apoptosis mediated by the mitochondrial permeability transition. Toxicol Sci 73(1):44–52
Bradham CA, Qian T, Streetz K, Trautwein C, Brenner DA, Lemasters JJ (1998) The mitochondrial permeability transition is required for tumor necrosis factor alpha-mediated apoptosis and cytochrome c release. Mol Cell Biol 18(11):6353–6364
Deng X, Yin F, Lu X, Cai B, Yin W (2006) The apoptotic effect of brucine from the seed of Strychnos nux-vomica on human hepatoma cells is mediated via Bcl-2 and Ca2+ involved mitochondrial pathway. Toxicol Sci 91(1):59–69
Szalai G, Krishnamurthy R, Hajnoczky G (1999) Apoptosis driven by IP3-linked mitochondrial calcium signals. EMBO J 18(22):6349–6361
Morita K, Kitayama T, Kitayama S, Dohi T (2006) Cyclic ADP-ribose requires FK506-binding protein to regulate intracellular Ca2+ dynamics and catecholamine release in acetylcholine-stimulated bovine adrenal chromaffin cells. J Pharmacol Sci 101(1):40–51
Mironov SL, Ivannikov MV, Johansson M (2005) [Ca2+]i signaling between mitochondria and endoplasmic reticulum in neurons is regulated by microtubules. From mitochondrial permeability transition pore to Ca2+-induced Ca2+ release. J Biol Chem 280(1):715–721
Campos CB, Paim BA, Cosso RG, Castilho RF, Rottenberg H, Vercesi AE (2006) Method for monitoring of mitochondrial cytochrome c release during cell death: immunodetection of cytochrome c by flow cytometry after selective permeabilization of the plasma membrane. Cytometry A 69(6):515–523
Mattson MP, Chan SL (2003) Calcium orchestrates apoptosis. Nat Cell Biol 5(12):1041–1043
Hoyt KR, Sharma TA, Reyndds I (1997) Trifluoperazine and dibucaine-induced inhibition of glutamate-induced mitochondria depolarization in rat cultured forebrain neurons. Br J Pharmacol 122(5):803–808
Dorrie J, Gerauer H, Wachter Y, Zunino SJ (2001) Resveratrol induces extensive apoptosis by depolarizing mitochondrial membranes and activating caspase-9 in acute lymphoblastic leukemia cells. Cancer Res 61(12):4731–4739
Whiteman M, Rose P, Siau JL, Cheung NS, Tan GS, Halliwell B, Armstrong JS (2005) Hypochlorous acid-mediated mitochondrial dysfunction and apoptosis in human hepatoma HepG2 and human fetal liver cells: role of mitochondrial permeability transition. Free Radic Biol Med 38(12):1571–1584
Jackson JG, Thayer SA (2006) Mitochondrial modulation of Ca2+-induced Ca2+-release in rat sensory neurons. J Neurophysiol 96(3):1093–1104
Dahlem YA, Wolf G, Siemen D, Horn TF (2006) Combined modulation of the mitochondrial ATP-dependent potassium channel and the permeability transition pore causes prolongation of the biphasic calcium dynamics. Cell Calcium 39(5):387–400
Acknowledgments
The study was supported by the Grant No. 30300455 of China Natural Science Foundation. The authors express thanks for the assistance from associate professor Ande Ma and Lianbo Chen for the excellent FACS analysis.
Author information
Authors and Affiliations
Corresponding author
Additional information
Xuemei Tian—Foundation item: Chinese National Natural Science Foundation (No.30300455).
Rights and permissions
About this article
Cite this article
Ma, X., Tian, X., Huang, X. et al. Resveratrol-induced mitochondrial dysfunction and apoptosis are associated with Ca2+ and mCICR-mediated MPT activation in HepG2 cells. Mol Cell Biochem 302, 99–109 (2007). https://doi.org/10.1007/s11010-007-9431-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-007-9431-8