胃癌p16基因启动子CpG岛甲基化研究

:100022790(2009)1821659203p16CpG,,,(,730000):2008211219;:2008212210:(2005404):.Tel:[phone]Email:[e-mail]:.(),.Tel:[phone]Email:[e-mail]islandatp16genepromoteringastriccarcinomasLUOChong,ZHOUYong2Ning,GUOXian,YANGYong2ChengDepartmentofGastroenterology,FirstHospital,LanzhouUniversity,Lanzhou730000,ChinaAbstractAIM:Toinvestigatetheexpressionofhypermethyla2tionofCpGislandatthep16genepromoteringastriccarcinomasanditssignificance.METHODS:HypermethylationofCpGislandatthep16genepromoterwasdeterminedbymethylation2specificpolymerasechainreaction(MSP)analysis.RESULTS:CpG2islandhypermethylaionwasnotdetectedincontrolbiopsyspecimensbutwasidentifiedinhumangastriccancercelllinesand74.1%(23/31)ofgastriccarcinomapatients.CpG2islandhypermethylaionofp16genewasdetectedin17adjacentgastrictissues(73.9%,17/23).Inprimarygastrictumors,nosignifi2cantassociationwasfoundbetweentumordifferentiationstatusandlymphnodemetastasisandCpGislandmethylationstatusofp16gene(P0.05,respectively).CONCLUSION:Hypermethyla2tionofCpGislandatthep16genepromoterisdetectedcommonlyingastriccarcinoma.p16promoterhypermethylationmaybeinvolvedintheearlygastriccarcinogenesisprocess.Keywordsp16;gastriccarcinoma;hypermethylation:p16CpG.:PCR(MSP)p16CpG.:CpG,CpG,23(74%)CpG.,17(74%)CpG.p16CpG(P0.05).:p16CpG,.p16;;R735.2A0CpG[1].,CpG.p16,[2].PCR(methylation2specificPCR,MSP)p16CpG,.11.1SGC27901BGC2823().2005210/200621031(22,9),2872(55).,.31:2,29;18,13;17,14.23.10().().PCR(GeneAmp2000)(BIO2RAD);(CHEMIGENIUS22BIOIMAGINESYSTEM);(PowerStation300);TaqDNApolymeraseDNA();EZDNAMethylation2GoldKitTM(ZYMORE2SEARCH).1.2CpG[3],:DNA;2gDNA,ZYMORESEARCH;(),:p162M,52TTATTAGAGGGTGGGGCGGATCGC23(sense),52GACCCCGAACCGCGACCGTAA23(anti2sense),150bp;p162U,52TATTAGAGGGT2GGGGTGGATTGT23(sense),52CAACCCCAAAC2CACAACCATAA23(antisense),151bp.9561(JFourthMilMedUniv)2009,30(18):SPSS15.0.2,P0.05.22.1p16MSPBGC2823SGC27901,(1,2).2.2p16MSP,(3,4).23p16CpG,74%(23/31),10(10/31,32%).17p16CpG,74%(17/23),8(8/23,35%).,p16CpG(2=0.000);,p16CpG(2=16.89,P0.01;2=15.24,P0.01).2.3p16CpG17/,11,65%;14,12,86%,/,CpG(2=1.769).18CpG72%(13/18),13CpG69%(9/13),,CpG(2=0.033).3,.,,DNA,,.CpG,CpG[1,3].,CpG,,,:(p16,p15),DNA(MLH1),(DAP2kinase,Caspase28),(E2cadherin,APC)[4-7].p16,[2].G1SDCDK4Rb,p16CDK4,G1.,p16CpG,p16CDK4,.MSPBGC2823SGC27901p16CpG,0661(JFourthMilMedUniv)2009,30(18)[8].,.[9-10],p16CpG.,Barrett(ER),APCp16CpG[11-12].E2cadherinCpG,CpG.,p16CpG73.9%,CpG,,p16,CpG[13-14].p16CpG,:p16CpG.,p16CpG.CpG,,,.CpG,CpG,.:.[1]TeodoridisJM,HardieC,BrownR.CpGislandmethylatorpheno2type(CIMP)incancer:causesandimplications[J].CancerLett,2008,268(2):177-186.[2]GuoX,MillsAA.p63,cellularsenescenceandtumordevelopment[J].CellCycle,2007,6(3):305-311.[3]ShamesDS,MinnaJD,GazdarAF.DNAmethylationinhealth,disease,andcancer[J].CurrMolMed,2007,7(1):85-102.[4]WangX,FengY,PanL,etal.TheproximalGC2richregionofp16(INK4a)genepromoterplaysaroleinitstranscriptionalregulation[J].MolCellBiochem,2007,301(122):259-266.[5]QuinaAS,BuschbeckM,DiCroceL.Chromatinstructureandepi2genetics[J].BiochemPharmacol,2006,72(11):1563-1569.[6]KawakamiK,BrabenderJ,LordRV,etal.HypermethylatedAPCDNAinplasmaandprognosisofpatientswithesophagealadenocarci2noma[J].JNatlCancerInst,2000,92(22):1805-1811.[7]LeeEJ,LeeBB,HanJ,etal.CpGislandhypermethylationofE2cadherin(CDH1)andintegrinalpha4isassociatedwithrecurrenceofearlystageesophagealsquamouscellcarcinoma[J].IntJCancer,2008,123(9):2073-2079.[8]BernalC,AguayoF,VillarroelC,etal.Reprimoasapotentialbio2markerforearlydetectioningastriccancer[J].ClinCancerRes,2008,14(19):6264-6269.[9]TanakaR,WangD,MorishitaY,etal.Lossoffunctionofp16geneandprognosisofpulmonaryadenocarcinoma[J].Cancer,2005,103(3):6082615.[10]NakataS,SugioK,UramotoH,etal.ThemethylationstatusandproteinexpressionofCDH1,p16(INK4A),andfragilehistidinetri2adinnonsmallcelllungcarcinoma:epigeneticsilencing,clinicalfeatures,andprognosticsignificance[J].Cancer,2006,106(10):2190-2199.[11]PetersFP,KaraMA,RosmolenWD,etal.Stepwiseradicalendo2scopicresectioniseffectiveforcompleteremovalofBarrettpsesopha2guswithearlyneoplasia:aprospectivestudy[J].AmJGastroen2terol,2006,101(7):1449-1457.[12]EadsCA,LordRV,KurumboorSK,etal.FieldsofaberrantCpGis2landhypermethylationinBarrett’sesophagusandassociatedadeno2carcinoma[J].CancerRes,2000,60(18):5021-5026.[13],,,.E2CdherinCpG[J].,2005,24:1220-1224.[14]SchildhausHU,KrockelI,LippertH,etal.Promoterhypermethyla2tionofp16INK4a,E2cadherin,O62MGMT,DAPKandFHITinad2enocarcinomasoftheesophagus,esophagogastricjunctionandproxi2malstomach[J].IntJOncol,2005,26(6):1493-1500.1661(JFourthMilMedUniv)2009,30(18)