Aberrant Promoter Methylation and Expression of UTF1 during Cervical Carcinogenesis 英文参考文献.docVIP

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Aberrant Promoter Methylation and Expression of UTF1 during Cervical Carcinogenesis 英文参考文献.doc

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Aberrant Promoter Methylation and Expression of UTF1 during Cervical Carcinogenesis 英文参考文献

AberrantPromoterMethylationandExpressionofUTF1 duringCervicalCarcinogenesis SamuelGuenin1*.,MustaphaMouallif1,2.,RachelDeplus3,XavierLampe3,NathalieKrusy1, EmilieCalonne3,KattyDelbecque4,FredericKridelka5,Franc?ois Fuks3,MyMustaphaEnnaji2, PhilippeDelvenne1 1Laboratory ofExperimental Pathology, Groupe Interdisciplinaire de Ge′noprote′omique Applique′e(GIGA)-Cancer, University of Lie`ge, Lie`ge, Belgium, 2Laboratory of Virology and Hygiene and Microbiology, University Hassan II-Mohammedia, Mohammedia, Morocco, 3Laboratory of Cancer Epigenetics, Free University of Brussels, Brussels,Belgium,4DepartmentofPathology,CHUSartTilman,Lie`ge, Belgium,5DepartmentofObstetricsandGynecology,CHUSartTilman,Lie`ge, Belgium Abstract Promotermethylationprofilesareproposedaspotentialprognosisand/ordiagnosisbiomarkersincervicalcancer.Upto now,littleisknownaboutthepromotermethylationprofileandexpressionpatternofstemcell(SC)markersduringtumor development.Inthisstudy,wewereinterestedtoidentifySCgenesmethylationprofilesduringcervicalcarcinogenesis.A genome-wide promoter methylation screening revealeda strong hypermethylation of Undifferentiated cell Transcription Factor1(UTF1)promoterincervicalcancerincomparisonwithnormalectocervix.BydirectbisulfitepyrosequencingofDNA isolatedfromliquid-basedcytologicalsamples,weshowedthatUTF1promotermethylationincreaseswithlesionseverity, the highest level of methylation being found in carcinoma. This hypermethylation was associated with increased UTF1 mRNA and protein expression. By using quantitative RT-PCR and Western Blot, we showed that both UTF1 mRNA and proteinarepresentinepithelialcancercelllines,evenintheabsenceofitstwomaindescribedregulatorsOct4AandSox2. Moreover,byimmunofluorescence,weconfirmedthenuclearlocalisationofUTF1incelllines.Surprisingly,directbisulfite pyrosequencing revealed that the inhibition of DNA methyltransferase by 5-aza-29-deoxycytidine was associated with decreasedUTF1genemethylationandexpressionintwocervicalcan