title: The contribution of N₂O₃ to the cytotoxicity of the nitric oxide donor DETA/NO: an emerging role for S-nitrosylation.
abstract: The relationship between the biological activity of NO and its chemistry is complex. The objectives of this study were to investigate the influence of oxygen tension on the cytotoxicity of the NO• donor DETA/NO and to determine the effects of oxygen tension on the key RNS (reactive nitrogen species) responsible for any subsequent toxicity. The findings presented in this study indicate that the DETA/NO-mediated cytotoxic effects were enhanced under hypoxic conditions. Further investigations revealed that neither ONOO⁻ (peroxynitrite) nor nitroxyl was generated. Fluorimetric analysis in the presence of scavengers suggest for the first time that another RNS, dinitrogen trioxide may be responsible for the cytotoxicity with DETA/NO. Results showed destabilization of HIF (hypoxia inducible factor)-1α and depletion of GSH levels following the treatment with DETA/NO under hypoxia, which renders cells more susceptible to DETA/NO cytotoxicity, and could account for another mechanism of DETA/NO cytotoxicity under hypoxia. In addition, there was significant accumulation of nuclear p53, which showed that p53 itself might be a target for S-nitrosylation following the treatment with DETA/NO. Both the intrinsic apoptotic pathway and the Fas extrinsic apoptotic pathway were also activated. Finally, GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is another important S-nitrosylated protein that may possibly play a key role in DETA/NO-mediated apoptosis and cytotoxicity. Therefore this study elucidates further mechanisms of DETA/NO mediated cytotoxicity with respect to S-nitrosylation that is emerging as a key player in the signalling and detection of DETA/NO-modified proteins in the tumour microenvironment.
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Ahlam A
authors: AA
authors: Ali
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Jonathan A
authors: JA
authors: Coulter
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Claire H
authors: CH
authors: Ogle
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Marie M
authors: MM
authors: Migaud
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: David G
authors: DG
authors: Hirst
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Tracy
authors: T
authors: Robson
authors: School of Pharmacy, McClay Research Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, U.K.
authors: Helen O
authors: HO
authors: McCarthy
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D015687:Cell Hypoxia; D045744:Cell Line, Tumor / Q000187:drug effects*; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000737:chemistry / Q000378:metabolism*; D009589:Nitrogen Oxides / Q000737:chemistry / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D026361:Reactive Nitrogen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014226:Triazenes / Q000737:chemistry; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000378:metabolism
chemical_list: C105351:1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D020030:Nitric Oxide Donors; D009589:Nitrogen Oxides; D026361:Reactive Nitrogen Species; D014226:Triazenes; D016159:Tumor Suppressor Protein p53; C031701:nitrogen trioxide; D009569:Nitric Oxide; D005978:Glutathione; D010100:Oxygen
references: Genes Dev. 2000 Jan 1;14(1):34-44
references: 10640274
references: Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):10958-63
references: 12177417
references: Nitric Oxide. 2010 May 15;22(4):259-65
references: 19818866
references: Nitric Oxide. 2008 Sep;19(2):68-72
references: 18503780
references: Mutat Res. 2001 Jun 2;477(1-2):97-110
references: 11376691
references: Cancer Lett. 2005 Aug 8;226(1):1-15
references: 16004928
references: World J Hepatol. 2010 Sep 27;2(9):337-44
references: 21161018
references: J Inorg Biochem. 2000 Apr;79(1-4):237-40
references: 10830872
references: Arch Biochem Biophys. 2009 Apr 15;484(2):114-6
references: 19393377
references: Am J Physiol Cell Physiol. 2002 Jul;283(1):C178-86
references: 12055086
references: Cancer Lett. 2010 Dec 8;298(2):204-11
references: 20674154
references: Anal Chem. 1998 Jul 1;70(13):2446-53
references: 9666719
references: Carcinogenesis. 2002 Jun;23(6):929-41
references: 12082014
references: Invest Ophthalmol Vis Sci. 2010 May;51(5):2575-86
references: 19959646
references: Proc Natl Acad Sci U S A. 2002 Aug 20;99(17):11127-32
references: 12177414
references: J Biol Chem. 1998 Jun 19;273(25):15646-53
references: 9624158
references: Biochem Pharmacol. 2010 Dec 1;80(11):1641-9
references: 20797387
references: Int J Cancer. 2009 Jan 1;124(1):223-32
references: 18924134
references: J Am Soc Mass Spectrom. 2011 May;22(5):888-97
references: 21472523
references: J Biol Chem. 1997 Jan 31;272(5):2841-5
references: 9006926
references: Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4671-6
references: 19261856
references: J Biol Chem. 2001 Aug 10;276(32):30085-91
references: 11404354
references: Expert Opin Ther Targets. 2012 Feb;16(2):167-78
references: 22233091
references: Chem Res Toxicol. 2007 Apr;20(4):709-14
references: 17388608
references: FEBS Lett. 1999 Feb 26;445(2-3):226-30
references: 10094462
references: Nat Cell Biol. 2008 Jul;10(7):866-73
references: 18552833
references: Oncol Rep. 2003 May-Jun;10(3):629-33
references: 12684635
references: PLoS Biol. 2012;10(5):e1001326
references: 22589701
references: Endocrinology. 2007 Sep;148(9):4352-61
references: 17540725
references: Gastroenterology. 2011 Jun;140(7):2009-18, 2018.e1-4
references: 21354149
references: J Biol Chem. 2002 May 3;277(18):16067-74
references: 11861645
references: Radiat Res. 1990 Apr;122(1):72-6
references: 2320727
references: Int J Oncol. 2000 Jan;16(1):109-17
references: 10601555
references: Eur J Biochem. 1995 Dec 1;234(2):472-8
references: 8536691
references: Curr Protoc Toxicol. 2001 May;Chapter 10:Unit 10.8
references: 20957636
references: Nitric Oxide. 1997 Feb;1(1):18-30
references: 9701041
references: Cell Mol Immunol. 2007 Oct;4(5):353-8
references: 17976315
references: J Neurochem. 2000 Oct;75(4):1455-64
references: 10987825
references: Science. 1999 Apr 23;284(5414):651-4
references: 10213689
references: Cancer Sci. 2005 Mar;96(3):157-63
references: 15771618
references: Int J Oncol. 2011 Apr;38(4):963-71
references: 21271218
references: J Neurosci. 1998 Jan 15;18(2):687-97
references: 9425011
references: Gen Pharmacol. 1998 Aug;31(2):179-86
references: 9688457
references: J Pharm Pharmacol. 2007 Jan;59(1):3-13
references: 17227615
references: J Biol Chem. 2003 May 2;278(18):15720-6
references: 12595536
references: J Gene Med. 2007 Jun;9(6):511-20
references: 17471586
references: Sci STKE. 2001 Jun 12;2001(86):pl1
references: 11752655
references: Physiol Rev. 2007 Jan;87(1):315-424
references: 17237348
references: Cell. 1994 Sep 23;78(6):931-6
references: 7923362
references: J Med Chem. 2006 Jul 13;49(14):4356-66
references: 16821795
references: Biochem Biophys Res Commun. 2005 Jun 10;331(3):718-25
references: 15865928
references: Cancer Res. 2007 Feb 15;67(4):1407-10
references: 17308075
references: Nat Cell Biol. 2010 Nov;12(11):1094-100
references: 20972425
references: Kidney Int. 2005 May;67(5):1890-8
references: 15840036
references: Biochemistry. 1998 Apr 21;37(16):5362-71
references: 9548918