title: MI-219-zinc combination: a new paradigm in MDM2 inhibitor-based therapy.
abstract: Zinc has a crucial role in the biology of p53 in that p53 binds to DNA through a structurally complex domain stabilized by zinc atom. The p53 negative regulator MDM2 protein also carries a C-terminal RING domain that coordinates two zinc atoms, which are responsible for p53 nuclear export and proteasomal degradation. In this clinically translatable study, we explored the critical role of zinc on p53 reactivation by MDM2 inhibitor, MI-219, in colon and breast cancer cells. ZnCl(2) enhanced MI-219 activity (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), apoptosis and colony formation), and chelation of zinc not only blocked the activity of MI-219, but also suppressed reactivation of the p53 and its downstream effector molecules p21(WAF1) and Bax. N,N,N'N'-tetrakis(-)[2-pyridylmethyl]-ethylenediamine (TPEN), a specific zinc chelator, but not 1,2-bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (Bapta-AM), a calcium chelator, blocked MI-219-induced apoptosis. Nuclear localization is a prerequisite for proper functioning of p53 and our results confirm that TPEN, and not Bapta-AM, could abrogate p53 nuclear localization and it interfered with p53 transcriptional activation. Addition of zinc suppressed the known p53 feedback MDM2 activation, which could be restored by TPEN. Co-immunoprecipitation studies verified that MI-219-mediated MDM2-p53 disruption could be suppressed by TPEN and restored by zinc. As such, single-agent therapies that target MDM2 inhibition, without supplemental zinc, may not be optimal in certain patients owing to the less recognized mild zinc deficiency among the 'at-risk population' as in the elderly who are more prone to cancers. Therefore, use of supplemental zinc with MI-219 will benefit the overall efficacy of MIs and this potent combination warrants further investigation.
authors.affiliation: Department of Pathology, Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Azmi
authors.forename: P A
authors.initials: PA
authors.lastname: Philip
authors.forename: F W J
authors.initials: FW
authors.lastname: Beck
authors.forename: Z
authors.initials: Z
authors.lastname: Wang
authors.forename: S
authors.initials: S
authors.lastname: Banerjee
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: D
authors.initials: D
authors.lastname: Yang
authors.forename: F H
authors.initials: FH
authors.lastname: Sarkar
authors.forename: R M
authors.initials: RM
authors.lastname: Mohammad
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000378:metabolism; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D002712:Chlorides / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000378:metabolism; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D045325:HCT116 Cells; D006801:Humans; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D017967:Zinc Compounds / Q000494:pharmacology*
chemical_list: D002614:Chelating Agents; D002712:Chlorides; D004791:Enzyme Inhibitors; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D017967:Zinc Compounds; C016837:zinc chloride; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
references.citation: Nutrition. 1993 May-Jun;9(3):218-24
references.pmid: 8353362
references.citation: J Biol Chem. 2009 Apr 24;284(17):11517-30
references.pmid: 19188367
references.citation: Science. 1994 Jul 15;265(5170):346-55
references.pmid: 8023157
references.citation: Biochem Biophys Res Commun. 2006 May 26;344(1):272-82
references.pmid: 16616005
references.citation: Genes Dev. 1993 Dec;7(12B):2565-74
references.pmid: 8276239
references.citation: J Lab Clin Med. 2001 Oct;138(4):250-6
references.pmid: 11574819
references.citation: Biochem J. 1993 Dec 1;296 ( Pt 2):403-8
references.pmid: 8257431
references.citation: J Nutr Biochem. 2004 Oct;15(10):572-8
references.pmid: 15542347
references.citation: Mol Carcinog. 1998 Mar;21(3):205-14
references.pmid: 9537652
references.citation: Biochemistry. 2007 Mar 13;46(10):2630-9
references.pmid: 17297920
references.citation: Mol Cell. 2006 Jul 21;23(2):251-63
references.pmid: 16857591
references.citation: Cell. 1997 Feb 7;88(3):323-31
references.pmid: 9039259
references.citation: Nat Rev Cancer. 2009 Dec;9(12):862-73
references.pmid: 19935675
references.citation: Prostate. 2004 Mar 1;58(4):374-81
references.pmid: 14968438
references.citation: Eur J Cancer. 2010 Apr;46(6):1122-31
references.pmid: 20156675
references.citation: Cell Signal. 1998 Mar;10(3):159-66
references.pmid: 9607138
references.citation: Am J Clin Nutr. 1998 Aug;68(2 Suppl):410S-413S
references.pmid: 9701153
references.citation: Cancer Cell. 2007 Oct;12(4):355-66
references.pmid: 17936560
references.citation: Chem Biol. 1994 Nov;1(3):153-61
references.pmid: 9383385
references.citation: Biochemistry. 2003 Mar 4;42(8):2396-403
references.pmid: 12600206
references.citation: J Med Chem. 2006 Jun 15;49(12):3432-5
references.pmid: 16759082
references.citation: Annu Rev Pharmacol Toxicol. 2009;49:223-41
references.pmid: 18834305
references.citation: Proc Natl Acad Sci U S A. 2002 Aug 6;99(16):10855-60
references.pmid: 12145320
references.citation: Met Ions Biol Syst. 2004;41:103-37
references.pmid: 15206115
references.citation: Trends Mol Med. 2007 Jan;13(1):23-31
references.pmid: 17126603
references.citation: Mol Cancer. 2009 Dec 03;8:115
references.pmid: 19958544
references.citation: Ann N Y Acad Sci. 1995 Sep 30;768:111-28
references.pmid: 8526340
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3933-8
references.pmid: 18316739
references.citation: Nutrition. 2001 Jul-Aug;17(7-8):685-7
references.pmid: 11448605
references.citation: Cell Cycle. 2008 Apr 1;7(7):842-7
references.pmid: 18414039
references.citation: J Med Chem. 2009 Dec 24;52(24):7970-3
references.pmid: 19928922
references.citation: Nature. 1997 May 15;387(6630):299-303
references.pmid: 9153396
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):2776-8
references.pmid: 9096291
references.citation: Toxicol Lett. 2006 Oct 25;166(3):222-8
references.pmid: 16930873
references.citation: J Lab Clin Med. 2002 Oct;140(4):272-89
references.pmid: 12389026
references.citation: Proc Nutr Soc. 1999 Aug;58(3):565-71
references.pmid: 10604188
references.citation: Adv Protein Chem. 1991;42:281-355
references.pmid: 1793007
references.citation: Br J Cancer. 2009 Sep 1;101(5):774-81
references.pmid: 19707204
references.citation: J Am Chem Soc. 2005 Jul 27;127(29):10130-1
references.pmid: 16028899
references.citation: Cell Death Differ. 2010 Jan;17(1):93-102
references.pmid: 19498444
references.citation: Nature. 2004 Feb 26;427(6977):789-90
references.pmid: 14985740
references.citation: Drug Resist Updat. 2003 Dec;6(6):313-22
references.pmid: 14744495
references.citation: Proc Natl Acad Sci U S A. 2002 Dec 24;99(26):16770-5
references.pmid: 12481036
references.citation: Biochem Pharmacol. 2010 Feb 15;79(4):565-74
references.pmid: 19788889
references.citation: Curr Opin Clin Nutr Metab Care. 2009 Nov;12(6):640-5
references.pmid: 19684515
references.citation: Mol Cell Biol. 2007 Feb;27(3):1056-68
references.pmid: 17116689
references.citation: Exp Biol Med (Maywood). 2006 May;231(5):611-8
references.pmid: 16636310
references.citation: Genes Dev. 1993 Dec;7(12B):2556-64
references.pmid: 8276238
references.citation: Mol Cancer Ther. 2008 Sep;7(9):2884-93
references.pmid: 18790769
references.citation: Food Nutr Bull. 2004 Mar;25(1 Suppl 2):S99-203
references.pmid: 18046856
references.citation: Oncogene. 2000 Nov 2;19(46):5227-36
references.pmid: 11077439