P38 MAPK inhibition enhancing ATO‐induced cytotoxicity against multiple myeloma cells

The resistance to arsenic trioxide (ATO) treatment is relatively common (55–80%) in multiple myeloma patients. This study found that ATO at clinically achievable concentrations (2–7 μmol/l) activated p38 mitogen‐activated protein kinase (MAPK) in both myeloma cell lines and primary myeloma cells, a finding not previously well‐documented in myeloma cells. Inhibition of p38 MAPK activation by pharmacological inhibitors (SB203580) or downregulation of p38 MAPK by siRNA significantly increased the apoptosis and/or growth inhibition induced by ATO treatment in myeloma cells. Combination of ATO and p38 MAPK inhibition abolished the interleukin‐6 enhanced protection of myeloma cells against ATO treatment. The ATO‐resistant cell line developed in our laboratory showed an increase in p38 MAPK activation. The increase of apoptosis by the combination of ATO and SB203580 was accompanied by the activation of caspase‐9 and caspase‐8 suggesting that both extrinsic and intrinsic apoptotic pathways are involved. Additionally, the p38 MAPK activation by ATO was associated with increased phosphorylation and upregulated expression of Heat shock protein 27. These results suggest that ATO‐induced p38 MAPK activation plays an important role in the resistance to ATO in myeloma cells and that p38 MAPK inhibition may overcome resistance to ATO treatment in myeloma patients.

[1]  Kenneth C. Anderson,et al.  P38 MAPK Inhibition Enhances PS-341 (bortezomib)-Induced Cytotoxicity Against Multiple Myeloma Cells. , 2004 .

[2]  Chen Dong,et al.  MAP kinases in the immune response. , 2002, Annual review of immunology.

[3]  S. Hilsenbeck,et al.  Arsenic trioxide-induced apoptosis in myeloma cells: p53-dependent G1 or G2/M cell cycle arrest, activation of caspase-8 or caspase-9, and synergy with APO2/TRAIL. , 2003, Blood.

[4]  M. Kizaki,et al.  Resveratrol induces apoptosis of human malignant B cells by activation of caspase-3 and p38 MAP kinase pathways. , 2006, Biochemical pharmacology.

[5]  N. Munshi,et al.  Hsp27 inhibits release of mitochondrial protein Smac in multiple myeloma cells and confers dexamethasone resistance. , 2003, Blood.

[6]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[7]  Masaharu Akiyama,et al.  Targeting p38 MAPK inhibits multiple myeloma cell growth in the bone marrow milieu. , 2003, Blood.

[8]  J Ross,et al.  Cardiac Muscle Cell Hypertrophy and Apoptosis Induced by Distinct Members of the p38 Mitogen-activated Protein Kinase Family* , 1998, The Journal of Biological Chemistry.

[9]  S. Minucci,et al.  Activation of Rac1 and the p38 Mitogen-activated Protein Kinase Pathway in Response to Arsenic Trioxide* , 2002, The Journal of Biological Chemistry.

[10]  A. Jemal,et al.  Cancer Statistics, 2007 , 2007, CA: a cancer journal for clinicians.

[11]  Louis W. Chang,et al.  Mitogen-activated protein kinases mediate arsenic-induced down-regulation of survivin in human lung adenocarcinoma cells , 2006, Archives of Toxicology.

[12]  Wei Tang,et al.  Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. , 1997, Blood.

[13]  T. Naoe,et al.  Sustained activation of c‐jun‐N‐terminal kinase plays a critical role in arsenic trioxide‐induced cell apoptosis in multiple myeloma cell lines , 2006, Cancer science.

[14]  E. Solary,et al.  Small heat shock proteins HSP27 and alphaB-crystallin: cytoprotective and oncogenic functions. , 2005, Antioxidants & redox signaling.

[15]  Jacques Landry,et al.  Inhibition of Daxx-Mediated Apoptosis by Heat Shock Protein 27 , 2000, Molecular and Cellular Biology.

[16]  L. Boise,et al.  Feasibility and correlates of arsenic trioxide combined with ascorbic acid-mediated depletion of intracellular glutathione for the treatment of relapsed/refractory multiple myeloma. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[17]  M. Hussein.  Trials of arsenic trioxide in multiple myeloma. , 2003, Cancer control : journal of the Moffitt Cancer Center.

[18]  A. Guimond,et al.  Characterization of 45-kDa/54-kDa HSP27 kinase, a stress-sensitive kinase which may activate the phosphorylation-dependent protective function of mammalian 27-kDa heat-shock protein HSP27. , 1995, European journal of biochemistry.

[19]  G. Kroemer,et al.  Heat shock proteins: endogenous modulators of apoptotic cell death. , 2001, Biochemical and biophysical research communications.

[20]  Yong Jiang,et al.  PRAK, a novel protein kinase regulated by the p38 MAP kinase , 1998, The EMBO journal.

[21]  H. Schaeffer,et al.  Mitogen-Activated Protein Kinases: Specific Messages from Ubiquitous Messengers , 1999, Molecular and Cellular Biology.

[22]  Michael E. Greenberg,et al.  Opposing Effects of ERK and JNK-p38 MAP Kinases on Apoptosis , 1995, Science.

[23]  Xinru Wang,et al.  Inhibition of mitogen‐activated protein kinase kinase enhances apoptosis induced by arsenic trioxide in human breast cancer MCF‐7 cells , 2005, Clinical and experimental pharmacology & physiology.

[24]  W. Fiers,et al.  The p38/RK mitogen‐activated protein kinase pathway regulates interleukin‐6 synthesis response to tumor necrosis factor. , 1996, The EMBO journal.

[25]  Constantine Mitsiades,et al.  p38 MAPK inhibition enhances PS-341 (bortezomib)-induced cytotoxicity against multiple myeloma cells , 2004, Oncogene.

[26]  B. Chauffert,et al.  HSP27 as a mediator of confluence-dependent resistance to cell death induced by anticancer drugs. , 1997, Cancer research.

[27]  L. Platanias.  Map kinase signaling pathways and hematologic malignancies. , 2003, Blood.

[28]  Guoqiang Chen,et al.  Arsenic trioxide and proteasome inhibitor bortezomib synergistically induce apoptosis in leukemic cells: the role of protein kinase Cδ , 2007, Leukemia.

[29]  K. Heidenreich,et al.  Apoptosis Induced by Withdrawal of Trophic Factors Is Mediated by p38 Mitogen-activated Protein Kinase* , 1997, The Journal of Biological Chemistry.

[30]  J. Landry,et al.  Induction of HSP27 phosphorylation and thermoresistance in Chinese hamster cells by arsenite, cycloheximide, A23187, and EGTA. , 1990, Radiation research.

[31]  C. Dominguez,et al.  p38 MAP kinase inhibitors: many are made, but few are chosen. , 2005, Current opinion in drug discovery & development.

[32]  T. Hideshima,et al.  Inhibition of p38α MAPK enhances proteasome inhibitor-induced apoptosis of myeloma cells by modulating Hsp27, Bcl-XL, Mcl-1 and p53 levels in vitro and inhibits tumor growth in vivo , 2006, Leukemia.

[33]  J. Landry,et al.  Regulation of actin filament dynamics by p38 map kinase-mediated phosphorylation of heat shock protein 27. , 1997, Journal of cell science.

[34]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[35]  Guido Kroemer,et al.  Hsp27 negatively regulates cell death by interacting with cytochrome c , 2000, Nature Cell Biology.

[36]  R. Morimoto,et al.  Role of the heat shock response and molecular chaperones in oncogenesis and cell death. , 2000, Journal of the National Cancer Institute.

[37]  N. Munshi,et al.  Arsenic trioxide inhibits growth of human multiple myeloma cells in the bone marrow microenvironment. , 2002, Molecular cancer therapeutics.

[38]  Qing‐Yu He,et al.  A proteome analysis of the arsenite response in cultured lung cells: evidence for in vitro oxidative stress-induced apoptosis. , 2004, The Biochemical journal.

[39]  W. Dalton,et al.  Ascorbic acid enhances arsenic trioxide-induced cytotoxicity in multiple myeloma cells. , 2001, Blood.

[40]  H. Koeffler,et al.  Induction of apoptosis by vitamin D3 analogue EB1089 in NCI‐H929 myeloma cells via activation of caspase 3 and p38 MAP kinase , 2000, British journal of haematology.

[41]  M. Lieberman,et al.  The MRP2/cMOAT Transporter and Arsenic-Glutathione Complex Formation Are Required for Biliary Excretion of Arsenic* , 2000, The Journal of Biological Chemistry.

[42]  L. Platanias,et al.  Role of the p38 mitogen-activated protein kinase pathway in the generation of arsenic trioxide-dependent cellular responses. , 2006, Cancer research.

[43]  H. Dombret,et al.  A clinical and pharmacological study of arsenic trioxide in advanced multiple myeloma patients , 2004, Leukemia.

[44]  K. Heidenreich,et al.  Insulin-like Growth Factor-I Induces bcl-2 Promoter through the Transcription Factor cAMP-Response Element-binding Protein* , 1999, The Journal of Biological Chemistry.

[45]  J. Landry,et al.  Oxidative stress-induced actin reorganization mediated by the p38 mitogen-activated protein kinase/heat shock protein 27 pathway in vascular endothelial cells. , 1997, Circulation research.

[46]  J. Santibañez,et al.  ERK 1,2 and p38 pathways are involved in the proliferative stimuli mediated by urokinase in osteoblastic SaOS‐2 cell line , 2001, Journal of cellular biochemistry.

[47]  W. Miller,et al.  JNK activation is a mediator of arsenic trioxide-induced apoptosis in acute promyelocytic leukemia cells. , 2004, Blood.

[48]  S. Waxman,et al.  Malignant cells can be sensitized to undergo growth inhibition and apoptosis by arsenic trioxide through modulation of the glutathione redox system. , 1999, Blood.

[49]  Minoru Takagi,et al.  Induction of Apoptosis by ASK1, a Mammalian MAPKKK That Activates SAPK/JNK and p38 Signaling Pathways , 1997, Science.

[50]  B. Barlogie,et al.  Clinical activity of arsenic trioxide for the treatment of multiple myeloma , 2002, Leukemia.

[51]  P. Fisher,et al.  mda-7 (IL-24) mediates selective apoptosis in human melanoma cells by inducing the coordinated overexpression of the GADD family of genes by means of p38 MAPK , 2002, Proceedings of the National Academy of Sciences of the United States of America.