title: Differential induction of reactive oxygen species through Erk1/2 and Nox-1 by FK228 for selective apoptosis of oncogenic H-Ras-expressing human urinary bladder cancer J82 cells.
abstract: OBJECTIVE
  This study sought to reveal mechanisms for differential regulation of reactive oxygen species (ROS) in histone deacetylase inhibitor FK228-induced selective apoptosis of oncogenic H-Ras-expressing human cancer cells.
  
  
  METHODS
  Human urinary bladder cancer J82 and oncogenic H-Ras-expressing J82 cells were used to reveal FK228-induced differential Erk1/2 activation, Nox-1 elevation, ROS production, glutathione (GSH) depletion, caspase activation, and apoptosis. Specific inhibitors were used to suppress Nox-1 activity and ROS production. Mek1/2 inhibitor was used to suppress Erk1/2 activation. Validated-specific siRNAs were used to knock down Nox-1. ROS levels, GSH levels, and caspase-3/7 activities were measured by GSH assay, flow cytometry and luminescence assays, respectively. Western blot analysis determined levels of Erk1/2 and Nox-1.
  
  
  RESULTS
  Erk1/2, Nox-1, ROS, caspase-3/7, and cell death were differentially induced, whereas GSH was differentially depleted by FK228 in oncogenic H-Ras-expressing J82 versus parental cells. Blockage of the ERK pathway resulted in suppressing oncogenic H-Ras- and FK228-induced Nox-1 elevation, ROS production, caspase activation, and cell death. Knockdown of Nox-1 by specific siRNAs reduced FK228-induced ROS production, caspase activation, and cell death.
  
  
  CONCLUSIONS
  Oncogenic H-Ras expression and FK228 treatment synergistically induced the ERK pathway, resulting in differentially increased Nox-1 elevation, ROS production, and GSH depletion, leading to differential caspase activation and cell death in oncogenic H-Ras-expressing J82 versus parental cells.
authors.affiliation: Anticancer Molecular Oncology Laboratory, Department of Comparative Medicine, College of Veterinary Medicine, The University of Tennessee, Knoxville, TN 37996, USA. schoudha@utk.edu
authors.forename: Shambhunath
authors.initials: S
authors.lastname: Choudhary
authors.forename: Kusum
authors.initials: K
authors.lastname: Rathore
authors.forename: Hwa-Chain Robert
authors.initials: HC
authors.lastname: Wang
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D047630:Depsipeptides / Q000494:pharmacology*; D004790:Enzyme Induction / Q000187:drug effects; D005978:Glutathione / Q000172:deficiency / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008928:Mitochondria / Q000378:metabolism; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases / Q000096:biosynthesis / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000096:biosynthesis*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000473:pathology
chemical_list: D000903:Antibiotics, Antineoplastic; D047630:Depsipeptides; D017382:Reactive Oxygen Species; C087123:romidepsin; D000074624:NADPH Oxidase 1; D019255:NADPH Oxidases; C402799:NOX1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020169:Caspases; D016283:Proto-Oncogene Proteins p21(ras); D005978:Glutathione
references.citation: J Biol Chem. 2004 Nov 12;279(46):47726-31
references.pmid: 15358777
references.citation: Int J Cancer. 2008 Apr 15;122(8):1757-64
references.pmid: 18076063
references.citation: Curr Top Cell Regul. 2000;36:151-80
references.pmid: 10842751
references.citation: Biol Chem. 2009 Mar;390(3):191-214
references.pmid: 19166318
references.citation: Exp Cell Res. 2006 Jul 1;312(11):2066-73
references.pmid: 16624288
references.citation: J Cancer Res Clin Oncol. 2009 Nov;135(11):1601-13
references.pmid: 19506904
references.citation: Mol Carcinog. 1999 Nov;26(3):180-8
references.pmid: 10559793
references.citation: Curr Biol. 2001 Apr 17;11(8):614-9
references.pmid: 11369207
references.citation: Nat Rev Cancer. 2005 Sep;5(9):713-25
references.pmid: 16110317
references.citation: Mol Cancer Ther. 2007 Mar;6(3):1099-111
references.pmid: 17363503
references.citation: J Biol Chem. 2002 May 31;277(22):19358-66
references.pmid: 11884408
references.citation: FEBS Lett. 2005 Jan 3;579(1):90-4
references.pmid: 15620695
references.citation: Cancer Sci. 2009 Aug;100(8):1382-8
references.pmid: 19493276
references.citation: Cancer Res. 2004 May 15;64(10):3580-5
references.pmid: 15150115
references.citation: Cancer Res. 2009 Apr 1;69(7):3157-64
references.pmid: 19293182
references.citation: Clin Cancer Res. 2004 Jun 1;10(11):3853-62
references.pmid: 15173094
references.citation: Oncogene. 2001 Mar 22;20(12):1486-96
references.pmid: 11313892
references.citation: J Biol Chem. 2007 Jun 15;282(24):17640-8
references.pmid: 17435218
references.citation: Pulm Pharmacol Ther. 2007;20(1):9-22
references.pmid: 16458553
references.citation: J Cell Sci. 2007 Mar 1;120(Pt 5):838-48
references.pmid: 17298981
references.citation: J Biochem Mol Biol. 2003 Jul 31;36(4):399-402
references.pmid: 12895299
references.citation: J Pharmacol Exp Ther. 2002 Mar;300(3):890-9
references.pmid: 11861795
references.citation: Am J Pathol. 2007 Dec;171(6):2021-32
references.pmid: 18055552
references.citation: Cancer Cell. 2006 Sep;10(3):241-52
references.pmid: 16959615
references.citation: Biochem J. 2000 Oct 1;351(Pt 1):183-93
references.pmid: 10998361
references.citation: Int J Cancer. 2000 Nov 15;88(4):620-5
references.pmid: 11058880
references.citation: Mol Carcinog. 1995 Jul;13(3):173-81
references.pmid: 7619220
references.citation: J Urol. 2003 Nov;170(5):1987-93
references.pmid: 14532839
references.citation: Oncogene. 2003 Mar 6;22(9):1349-57
references.pmid: 12618760
references.citation: J Cancer Res Clin Oncol. 2007 Oct;133(10):725-39
references.pmid: 17487507
references.citation: J Biol Chem. 1999 Jun 4;274(23):16188-97
references.pmid: 10347173
references.citation: Oncogene. 2008 Aug 21;27(36):4921-32
references.pmid: 18454176