Differential Methylation of the HPV 16 Upstream Regulatory Region during Epithelial Differentiation and Neoplastic Transformation 英文参考文献.docVIP

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Differential Methylation of the HPV 16 Upstream Regulatory Region during Epithelial Differentiation and Neoplastic Transformation 英文参考文献.doc

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Differential Methylation of the HPV 16 Upstream Regulatory Region during Epithelial Differentiation and Neoplastic Transformation 英文参考文献

DifferentialMethylationoftheHPV16Upstream RegulatoryRegionduringEpithelialDifferentiationand NeoplasticTransformation SvetlanaVinokurova,MagnusvonKnebelDoeberitz* DepartmentofAppliedTumorBiology,InstituteofPathology,UniversityofHeidelberg,Heidelberg,Germany Abstract High risk human papillomaviruses are squamous epitheliotropic viruses that may cause cervical and other cancers. HPV replication depends on squamous epithelial differentiation. Transformation of HPV-infected cells goes along with substantialalterationoftheviralgeneexpressionprofileandpreferentiallyoccursattransformationzonesusuallyatthe uterinecervix.Methylationoftheviralgenomemayaffectregulatoryfeaturesthatcontroltranscriptionandreplicationof theviralgenome.Therefore,weanalyzedthemethylationpatternoftheHPV16upstreamregulatoryregion(URR)during squamousepithelialdifferentiationandneoplastictransformationandanalyzedhowshiftsintheHPVURRmethylomemay affectviralgeneexpressionandreplication.HPV16positivebiopsysectionsencompassingallstagesofanHPVinfection (latent,permissiveandtransforming)weremicro-dissectedandDNAwasisolatedfromcellfractionsrepresentingthebasal, intermediate,andsuperficialcelllayers,each,aswellasfromtransformedp16INK4a-positivecells.Weobservedfundamental changesinthemethylationprofileoftranscriptionfactorbindingsitesintheHPV16upstreamregulatoryregionlinkedto thesquamousepithelialdifferentiationstage.Squamousepithelialtransformationindicatedbyp16INK4aoverexpressionwas associatedwithmethylationofthedistalE2bindingsite1leadingtohyper-activationoftheHPV16URR.Adjacentnormal but HPV 16-infected epithelial areas retained hyper-methylated HPV DNA suggesting that these viral genomes were inactivated. These data suggest that distinct shifts of the HPV 16 methylome are linked to differentiation dependent transcriptionandreplicationcontrolandmaytriggerneoplastictransformation. Citation:VinokurovaS,vonKnebelDoeberitzM(2011)DifferentialMethylationoftheHPV16UpstreamRegulatoryRegionduringEpithelialDifferenti