Allele-Specific Regulation of Matrix Metalloproteinase-3 Gene by Transcription Factor NFκB 英文参考文献.docVIP

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Allele-Specific Regulation of Matrix Metalloproteinase-3 Gene by Transcription Factor NFκB 英文参考文献.doc

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Allele-Specific Regulation of Matrix Metalloproteinase-3 Gene by Transcription Factor NFκB 英文参考文献

Allele-SpecificRegulationofMatrixMetalloproteinase-3 GenebyTranscriptionFactorNFkB VeronikaSouslova1,PaulA.Townsend2,JelenaMann3,ChrisM.vanderLoos4,AnnaMotterle1 ,Fulvio D’Acquisto1,DerekA.Mann3,ShuYe1* 1William Harvey Research Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom, 2Human Genetics Division, MP808, School of Medicine, University of Southampton, Southampton, United Kingdom, 3Institute of Cellular Medicine, Newcastle University, Newcastle,UnitedKingdom,4DepartmentofPathology,AcademicMedicalCentre,Amsterdam,TheNetherlands Abstract Background: Matrix metalloproteinase-3 (MMP3) is implicated in the pathogenesis and progression of atherosclerotic lesions. Previous studies suggested that MMP3 expression is influenced by a polymorphism (known as the 5A/6A polymorphism)inthepromoteroftheMMP3geneandthatthispolymorphismislocatedwithinacis-elementthatinteracts with the transcription factor NFkB. In the present study, we sought to investigate whether MMP3 and NFkB were co- localized in atherosclerotic lesions and whether NFkB had differential effects on the two alleles of the MMP3 5A/6A polymorphism. Methodology/PrincipalFindings:ImmunohistochemicalexaminationshowedthatMMP3andboththeNFkBp50andp65 subunitswereexpressedabundantlyinmacrophagesinatheroscleroticlesionsandthatMMP3expressionwasco-localized with p50 and p65. Chromatin immunoprecipitation experiments showed interaction of p50 and p65 with the MMP3 promoterinmacrophages,withgreaterbindingtothe5Aallelethantothe6Aallele.Reportergeneassaysintransiently transfected macrophages showed that the 5A allele had greater transcriptional activity than the 6A allele, and that this allele-specific effect was augmented when the cells were treated with the NFkB activator lipopolysaccharides or co- transfectedwithp50and/orp65expressingplasmids,butwasreducedwhenthecellsweretreatedwiththeNFk Binhibitor 6-Amino-4-(4-phenoxyphenylethylamino)-quinazolineort