title: STAT3 regulates MMP3 in heme-induced endothelial cell apoptosis.
abstract: BACKGROUND
  We have previously reported that free Heme generated during experimental cerebral malaria (ECM) in mice, is central to the pathogenesis of fatal ECM. Heme-induced up-regulation of STAT3 and CXCL10 promotes whereas up-regulation of HO-1 prevents brain tissue damage in ECM. We have previously demonstrated that Heme is involved in the induction of apoptosis in vascular endothelial cells. In the present study, we further tested the hypothesis that Heme reduces blood-brain barrier integrity during ECM by induction of apoptosis of brain vascular endothelial cells through STAT3 and its target gene matrix metalloproteinase three (MMP3) signaling.
  
  
  METHODS
  Genes associated with the JAK/STAT3 signaling pathway induced upon stimulation by Heme treatment, were assessed using real time RT(2) Profile PCR arrays. A human MMP3 promoter was cloned into a luciferase reporter plasmid, pMMP3, and its activity was examined following exposure to Heme treatment by a luciferase reporter gene assay. In order to determine whether activated nuclear protein STAT3 binds to the MMP3 promoter and regulates MMP3 gene, we conducted a ChIP analysis using Heme-treated and untreated human brain microvascular endothelial cells (HBVEC), and determined mRNA and protein expression levels of MMP3 using qRT-PCR and Western blot. Apoptosis in HBVEC treated with Heme was evaluated by MTT and TUNEL assay.
  
  
  RESULTS
  The results show that (1) Heme activates a variety of JAK/STAT3 downstream pathways in HBVEC. STAT3 targeted genes such as MMP3 and C/EBPb (Apoptosis-related genes), are up regulated in HBVEC treated with Heme. (2) Heme-induced HBVEC apoptosis via activation of STAT3 as well as its downstream signaling molecule MMP3 and upregulation of CXCL10 and HO-1 expressions. (3) Phosphorylated STAT3 binds to the MMP3 promoter in HBVEC cells, STAT3 transcribed MMP3 and induced MMP3 protein expression in HBVEC cells.
  
  
  CONCLUSIONS
  Activated STAT3 binds to the MMP3 promoter region and regulates MMP3 in Heme-induced endothelial cell apoptosis.
authors: Department of Microbiology, Biochemistry and Immunology, Morehouse School of Medicine, Atlanta, Georgia, USA. mliu@msm.edu
authors: Mingli
authors: M
authors: Liu
authors: Nana O
authors: NO
authors: Wilson
authors: Jacqueline M
authors: JM
authors: Hibbert
authors: Jonathan K
authors: JK
authors: Stiles
mesh_terms: D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D042783:Endothelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling; D006418:Heme / Q000378:metabolism*; D006801:Humans; D053612:Janus Kinases / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D015533:Transcriptional Activation
chemical_list: D050796:STAT3 Transcription Factor; D006418:Heme; D053612:Janus Kinases; D019278:Matrix Metalloproteinase 3
references: PLoS One. 2012;7(8):e43313
references: 22905257
references: Am J Pathol. 2011 Mar;178(3):1316-28
references: 21356382
references: Microcirculation. 2007 Aug;14(6):593-602
references: 17710630
references: Antioxid Redox Signal. 2006 Jul-Aug;8(7-8):1153-63
references: 16910763
references: Am J Trop Med Hyg. 2009 Apr;80(4):541-6
references: 19346372
references: J Neurochem. 2006 Sep;98(5):1353-68
references: 16923154
references: Toxicology. 2011 Feb 27;280(3):126-34
references: 21146578
references: Cell. 2011 Apr 29;145(3):398-409
references: 21529713
references: Blood. 2009 Nov 5;114(19):4243-52
references: 19713460
references: Int J Exp Pathol. 1998 Dec;79(6):369-91
references: 10319019
references: J Neuroinflammation. 2012 Mar 06;9:46
references: 22394415
references: Am J Physiol Cell Physiol. 2003 May;284(5):C1309-18
references: 12676658
references: Am J Transplant. 2005 Aug;5(8):1875-85
references: 15996234
references: Development. 2005 Nov;132(21):4675-85
references: 16192306
references: J Exp Med. 2000 Oct 16;192(8):1205-11
references: 11034611
references: PLoS One. 2012;7(3):e34280
references: 22479586
references: Brain. 2011 Apr;134(Pt 4):1127-39
references: 21371995
references: Annu Rev Cell Dev Biol. 2001;17:463-516
references: 11687497
references: Mol Cell Biol. 2004 Jun;24(12):5421-33
references: 15169904
references: J Inflamm (Lond). 2011 Mar 07;8:7
references: 21385378
references: PLoS Pathog. 2010 May 20;6(5):e1000916
references: 20502682
references: Cell Physiol Biochem. 2011;28(1):115-24
references: 21865854
references: Eur J Immunol. 2009 Jul;39(7):1850-63
references: 19585512
references: Mol Cell Biol. 1998 May;18(5):2553-8
references: 9566875
references: Infect Immun. 2006 Jun;74(6):3262-70
references: 16714553
references: Heart Fail Rev. 2011 Jan;16(1):35-47
references: 20407820
references: J Pharmacol Sci. 2011;117(4):213-22
references: 22056652
references: Mol Cell Biol. 2008 Apr;28(7):2391-413
references: 18172013
references: Inflammation. 2013 Oct;36(5):1064-74
references: 23612802
references: Int J Biochem Cell Biol. 2009 Apr;41(4):711-6
references: 18930163
references: Pharmacol Ther. 2008 Nov;120(2):172-85
references: 18786563
references: Am J Respir Cell Mol Biol. 1996 Jul;15(1):141-7
references: 8679218
references: Neuropathol Appl Neurobiol. 2007 Oct;33(5):560-71
references: 17442059
references: PLoS One. 2011 Mar 03;6(3):e14741
references: 21390226
references: Cell. 1999 Jul 23;98(2):137-46
references: 10428026
references: Mol Endocrinol. 2008 Dec;22(12):2677-88
references: 18927239
references: Cell. 1999 Aug 6;98(3):295-303
references: 10458605
references: J Immunol. 2006 Dec 1;177(11):7880-8
references: 17114459
references: FASEB J. 1995 Jul;9(10):899-909
references: 7542214
references: Oncogene. 2002 Mar 27;21(13):2000-8
references: 11960372
references: Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):177-82
references: 16371473
references: J Biol Chem. 2010 Nov 19;285(47):36842-8
references: 20855888
references: Mol Biochem Parasitol. 2008 Dec;162(2):172-6
references: 18848585
references: J Infect Dis. 2003 Apr 15;187(8):1283-90
references: 12696008
references: Prog Neurobiol. 2010 Jun;91(2):140-51
references: 20117166
references: J Neuroinflammation. 2010 Sep 09;7:54
references: 20828402
references: Am J Physiol Cell Physiol. 2007 Mar;292(3):C1123-31
references: 17035297
references: Nat Med. 2007 Jun;13(6):703-10
references: 17496899
references: Trop Med Int Health. 2009 Jul;14(7):810-6
references: 19413744
references: Mol Biochem Parasitol. 2001 Jul;115(2):129-43
references: 11420100
references: Am J Physiol Renal Physiol. 2007 Oct;293(4):F1222-30
references: 17652371
references: J Biol Chem. 1999 Jul 30;274(31):21491-4
references: 10419448
references: J Exp Med. 1993 Jan 1;177(1):145-53
references: 8418196
references: PLoS One. 2012;7(8):e43610
references: 22952718
references: Acta Trop. 2008 Jan;105(1):10-5
references: 17854755
references: Nat Rev Cancer. 2004 Feb;4(2):97-105
references: 14964307
references: Am J Pathol. 2006 Oct;169(4):1390-401
references: 17003494
references: Endocrinology. 2005 May;146(5):2229-38
references: 15661856
references: Cell Mol Immunol. 2008 Oct;5(5):341-8
references: 18954557
references: Glia. 2010 Nov 15;58(15):1858-70
references: 20737478
references: Infect Immun. 2013 Jul;81(7):2448-58
references: 23630962
references: Biochem Biophys Res Commun. 2010 Dec 3;403(1):30-5
references: 21036157
references: J Neurosci Res. 2010 Nov 1;88(14):3090-101
references: 20568281
references: Liver Transpl. 2010 May;16(5):600-10
references: 20440769
references: Cell Host Microbe. 2007 Aug 16;2(2):130-8
references: 18005727
references: Nat Med. 2007 Jun;13(6):667-9
references: 17554329
references: Basic Res Cardiol. 2007 Sep;102(5):393-411
references: 17918316
references: Blood. 2003 Apr 1;101(7):2850-7
references: 12517811
references: J Cereb Blood Flow Metab. 2010 Dec;30(12):1939-50
references: 20354546
references: Pediatr Res. 2010 Oct;68(4):267-74
references: 20606600
references: Science. 2002 May 31;296(5573):1653-5
references: 12040185
references: J Neuroimmune Pharmacol. 2010 Sep;5(3):370-86
references: 20401700
references: Infect Immun. 2012 Apr;80(4):1445-54
references: 22290142