Glibenclamide exerts an antitumor activity through reactive oxygen species-c-jun NH2-terminal kinase pathway in human gastric cancer cell line MGC-803.

Glibenclamide, a blocker of ATP-sensitive potassium (K(ATP)) channels, can suppress progression of many cancers, but the involved mechanism is unclear. Herein we reported that MGC-803 cells expressed the K(ATP) channels composed of Kir6.2 and SUR1 subunits. Glibenclamide induced cellular viability decline, coupled with cell apoptosis and reactive oxygen species (ROS) generation in MGC-803 cells. Meanwhile, glibenclamide increased NADPH oxidase catalytic subunit gp91(phox) expression and superoxide anion (O2-) generation, and caused mitochondrial respiration dysfunction in MGC-803 cells, suggesting that glibenclamide induced an increase of ROS derived from NADPH oxidase and mitochondria. Glibenclamide could also lead to loss of mitochondrial membrane potential, release of cytochrome c and apoptosis-inducing factor (AIF), and activation of c-jun NH2-terminal kinase (JNK) in MGC-803 cells. Pretreatment with antioxidant N-acetyl-l-cysteine (NAC) prevented glibenclamide-induced JNK activation, apoptosis and cellular viability decline. Furthermore, glibenclamide greatly decreased the cellular viability, induced apoptosis and inhibited Akt activation in wild-type mouse embryonic fibroblast (MEF) cells but not in JNK1-/- or JNK2-/- MEF cells. Taken together, our study reveals that glibenclamide exerts an antitumor activity in MGC-803 cells by activating ROS-dependent, JNK-driven cell apoptosis. These findings provide insights into the use of glibenclamide in the treatment of human gastric cancer.

[1]  Rodrigo Gc,et al.  ATP-sensitive potassium channels. , 2005, Current pharmaceutical design.

[2]  Surong Zhang,et al.  In vitro effect of manganese chloride exposure on reactive oxygen species generation and respiratory chain complexes activities of mitochondria isolated from rat brain. , 2004, Toxicology in vitro : an international journal published in association with BIBRA.

[3]  S. Orrenius,et al.  Triggering and modulation of apoptosis by oxidative stress. , 2000, Free radical biology & medicine.

[4]  L. Jan,et al.  Merging functional studies with structures of inward-rectifier K+ channels , 2003, Nature Reviews Neuroscience.

[5]  J. Downward.  PI 3-kinase, Akt and cell survival. , 2004, Seminars in cell & developmental biology.

[6]  R. Gottlieb,et al.  Nitrogen cavitation for cell disruption to obtain mitochondria from cultured cells. , 2000, Methods in enzymology.

[7]  Sébastien Bonnet,et al.  A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth. , 2007, Cancer cell.

[8]  J. McCord.  Human disease, free radicals, and the oxidant/antioxidant balance. , 1993, Clinical biochemistry.

[9]  D. Bar-Sagi,et al.  Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. , 2000, Science.

[10]  M. Oz,et al.  Improvement of myocardial mitochondrial function after hemodynamic support with left ventricular assist devices in patients with heart failure. , 1998, The Journal of thoracic and cardiovascular surgery.

[11]  F. Ashcroft.  ATP-sensitive potassium channelopathies: focus on insulin secretion. , 2005, The Journal of clinical investigation.

[12]  Y. Wong,et al.  Molecular mechanisms mediating the G protein-coupled receptor regulation of cell cycle progression , 2007, Journal of molecular signaling.

[13]  N. Kruger,et al.  The bradford method for protein quantitation. , 1988, Methods in molecular biology.

[14]  B. Vanhaesebroeck,et al.  The PI3K-PDK1 connection: more than just a road to PKB. , 2000, The Biochemical journal.

[15]  D. Leibfritz,et al.  Free radicals and antioxidants in normal physiological functions and human disease. , 2007, The international journal of biochemistry & cell biology.

[16]  João Wosniak,et al.  Analysis of DHE-derived oxidation products by HPLC in the assessment of superoxide production and NADPH oxidase activity in vascular systems. , 2007, American journal of physiology. Cell physiology.

[17]  Colin G. Nichols,et al.  KATP channels as molecular sensors of cellular metabolism , 2006, Nature.

[18]  Baofeng Yang,et al.  HERG K+ channel, a regulator of tumor cell apoptosis and proliferation. , 2002, Cancer research.

[19]  Michael Karin,et al.  Reactive Oxygen Species Promote TNFα-Induced Death and Sustained JNK Activation by Inhibiting MAP Kinase Phosphatases , 2005, Cell.

[20]  D. Spray,et al.  KATP Channels Regulate Mitogenically Induced Proliferation in Primary Rat Hepatocytes and Human Liver Cell Lines , 2000, The Journal of Biological Chemistry.

[21]  Xiao-Ming Yin,et al.  Cyanidin-3-rutinoside, a Natural Polyphenol Antioxidant, Selectively Kills Leukemic Cells by Induction of Oxidative Stress* , 2007, Journal of Biological Chemistry.

[22]  S. Seino,et al.  Roles of KATP channels as metabolic sensors in acute metabolic changes. , 2005, Journal of molecular and cellular cardiology.

[23]  R. Lotan,et al.  N-(4-Hydroxyphenyl)retinamide-induced apoptosis triggered by reactive oxygen species is mediated by activation of MAPKs in head and neck squamous carcinoma cells , 2006, Oncogene.

[24]  K. Davies.  Oxidative stress: the paradox of aerobic life. , 1995, Biochemical Society symposium.

[25]  D. Severson,et al.  Modulation of potassium currents by angiotensin and oxidative stress in cardiac cells from the diabetic rat , 2005, The Journal of physiology.

[26]  J C Reed,et al.  Mitochondria and apoptosis. , 1998, Science.

[27]  B. Hemmings,et al.  Physiological roles of PKB/Akt isoforms in development and disease. , 2007, Biochemical Society transactions.

[28]  Jinsong Liu,et al.  Selective killing of oncogenically transformed cells through a ROS-mediated mechanism by beta-phenylethyl isothiocyanate. , 2006, Cancer cell.

[29]  Shivendra V. Singh,et al.  c-Jun NH2-Terminal Kinase Signaling Axis Regulates Diallyl Trisulfide–Induced Generation of Reactive Oxygen Species and Cell Cycle Arrest in Human Prostate Cancer Cells , 2006 .

[30]  J. Holtz,et al.  Induction of NAD(P)H Oxidase by Oxidized Low-Density Lipoprotein in Human Endothelial Cells: Antioxidative Potential of Hydroxymethylglutaryl Coenzyme A Reductase Inhibitor Therapy , 2001, Circulation.

[31]  B. Kalyanaraman,et al.  Detection of 2-hydroxyethidium in cellular systems: a unique marker product of superoxide and hydroethidine , 2007, Nature Protocols.

[32]  H. Pelicano,et al.  Novel action of paclitaxel against cancer cells: bystander effect mediated by reactive oxygen species. , 2007, Cancer research.

[33]  J. Lambeth.  NOX enzymes and the biology of reactive oxygen , 2004, Nature Reviews Immunology.

[34]  H. Nakamura,et al.  Tumor-targeted induction of oxystress for cancer therapy , 2007, Journal of drug targeting.

[35]  T. Herdegen,et al.  JNK2 Translocates to the Mitochondria and Mediates Cytochrome c Release in PC12 Cells in Response to 6-Hydroxydopamine* , 2004, Journal of Biological Chemistry.

[36]  B. O’Rourke,et al.  Mitochondrial K(ATP) channels in cell survival and death. , 2005, Journal of molecular and cellular cardiology.

[37]  P. Vandenabeele,et al.  Toxic proteins released from mitochondria in cell death , 2004, Oncogene.

[38]  K. Okumura,et al.  Reactive oxygen species mediate crosstalk between NF-κB and JNK , 2006, Cell Death and Differentiation.

[39]  J. Woodgett,et al.  Functional distinctions of protein kinase B/Akt isoforms defined by their influence on cell migration. , 2006, Trends in cell biology.

[40]  Qing‐Yu He,et al.  Gold(III) Porphyrin 1a Induced Apoptosis by Mitochondrial Death Pathways Related to Reactive Oxygen Species , 2005 .

[41]  M. Conti.  Targeting K+ channels for cancer therapy. , 2004, Journal of experimental therapeutics & oncology.

[42]  P. Maher,et al.  The Regulation of Reactive Oxygen Species Production during Programmed Cell Death , 1998, The Journal of cell biology.

[43]  Mansoor Abdul,et al.  Expression and activity of potassium ion channels in human prostate cancer. , 2002, Cancer letters.

[44]  R. Coombes,et al.  Paclitaxel-induced nuclear translocation of FOXO3a in breast cancer cells is mediated by c-Jun NH2-terminal kinase and Akt. , 2006, Cancer research.

[45]  Sten Orrenius,et al.  Mitochondrial oxidative stress: implications for cell death. , 2007, Annual review of pharmacology and toxicology.

[46]  P. Schumacker,et al.  Reactive oxygen species in cancer cells: live by the sword, die by the sword. , 2006, Cancer cell.

[47]  J. Suttles,et al.  Membrane Potassium Channels and Human Bladder Tumor Cells: II. Growth Properties , 1998, The Journal of Membrane Biology.

[48]  D. Busija,et al.  Targeting mitochondrial ATP-sensitive potassium channels—a novel approach to neuroprotection , 2004, Brain Research Reviews.

[49]  Z. Ungvari,et al.  Chronic high pressure-induced arterial oxidative stress: involvement of protein kinase C-dependent NAD(P)H oxidase and local renin-angiotensin system. , 2004, The American journal of pathology.