血浆p16基因异常甲基化检测在肺癌临床诊断中的价值

©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(2009)01-0076-03p16,:p16,DNA,,PCR(MSP)5132p16,62.7%,p16:;p16;;:R73412:ADiagnosticValueofDetectionofAberrantlyMethylatedp16inPlasmaofPatientswithLungCancerMAXin-ping,WUYi(DepartmentofClinicalLabortory,HunanProvincialPeoplesHospital,Changsha410005,Hunan)Abstract:ObjectiveToinvestigatethep16genepromoteraberrantmethylationintheplasmaofpatientswithlungcanceranditspossibleclinicalapplicationinmoleculardiagnosisoflungcancer.MethodsFreeDNAwasextractedfromperipheralbloodplasmaoflungcancerpatients.Aftermodifyingwithsodiumbisulfiteandpurifying,thentheywereusedastemplateformethylationspecificPCR(MSP).Resultsp16genepromoteraberrantmethylationwasfoundin32of51samplesoflungcancerpatients(62.7%)usingmethylationspecificPCR,nomethylatedp16genewasdetectedincontrolsamples.ConclusionsThemeasurementofmethylatedp16genemaybehelpfulinearlydiagnosisoflungcancer.Keywords:Lungcancer;p16;Methylation;Plasma:(410005):,,,:UCP2UCP2ROS,[][1]HionaA,LeeuwenburghC.TheroleofmitochondrialDNAmutationsinagingandsarcopen-ia:implicationsforthemitochondrialcycletheo2ryofaging[J].ExpGerontol,2008,43(1):24-33.[2]DhamratSS,StephensJW,CooperJA,etal.Cardiovascularriskinhealthymenandmarkersofoxidativestressindiabeticmenareassoci2atedwithcommonvariationinthegeneforuncouplingportein2[J].EurHeartJ,2004,25(6):468-475.[3]EchtayKS,PakayJL,EstevesTC,BrandMD.Hydroxynonenalanduncouplingproteins:amodelforprotectionagainstoxidativedamage[J].Biofactors,2005,24(1-4):119-130.[4]FulopP,DerdakZ,SheetsA,etal.LackofUCP2reducesFasmediat2edliverinjuryinob/obmiceandrevealsimportanceofcellspecificUCP2expression[J].Hepatology,2006,44(3):592-601.[5]VoglerS,PahnkeJ,RoussetS,etal.UncouplingProtein2HasPro2tectiveFunctionduringExperimentalAutoimmuneEncephalomyelitis[J].AmJPathol,2006,168(5):1570-1575.[6]LeeKU,LeeIK,HanJ,etal.Effectsofrecombinantadenovirus-mediateduncouplingprotein2overexpressiononendothelialfunctionandapoptosis[J].CircRes,2005,96(11):1200-1207.[7]EchtayKS,EstevesTC,PakayJL,etal.Asignalingrolefor4-hy2droxy-2-nonenalinregulationofmitochondrialuncoupling[J].EM2BOJ,2003,4103-4110.[8]MattiassonG,SullivanPG.TheemergingfunctionsofUCP2inheath,disease,andtherapeutics[J].AntioxidRedoxSigal,2006,8(1-2):1-38.[9]DeyA,CederbaumAI.Ethanol-inducedoxidativestressplaysamajorroleinthemechanismsbywhichethanolproducesliverinjury[J].Hep2atology,2006,43(2):63-74.[10]KussmaulL,HirstJ.ThemechanismofsuperoxideproductionbyNADH:ubiquinoneoxidoreductase(complex)frombovineheartmi2tochondria[J].ProcNatlAcadSciUSA,2006,103(20):7607-7612.[11]HorimotoM,FulopP,DerdakZ,etal.Uncouplingprotein2defi2ciencypromotesoxidantstressanddelayslivesregenerationinmice[J].Hepatology,2004,39(2):386-392.[12]BrandMD,EstevesTC.Physiologicalfunctionsofthemitochdeialuncouplingproteins2and3[J].CellMetab,2005,2(2):85-93.[13]IshizawaM,MizushigeK,NomaT,etal.Anantioxidanttreatmentpotentiallyprotectsmyocardialenergymetabolismbyrequlatinguncou2plingprotein2expressioninachronicbeta-adrenergicstimulationratmodel[J].LifeSei,2006,78(25):2974-2982.[14]BrookesPS.MitochondrialH(+)leakandROSgeneration:anoddcouple[J].FreeRadicBiolMed,2005,38(1):12-23.(:2008-12-13)6720092161PracticalPreventiveMedicine,Feb.2009,Vol16,No.1©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.[1],,[2]p16,,,[3]PCR(MSP)p16,11.11.1.151,,34,17,4375,5530,19,11,2260,531.1.2DNAQIAampDNABloodMiniKitHotStarTaqQIAGEN,WizardDNAClean-UpSystemPromega,SigmaInvitrogen,:M:5-TTATTAGAGGGTGGGGCGGATCGC-3,M:5-GACCC2CGAACCGCGACCCTAA-3,U:5-TTATTA2GAGGGTGGGGTGGATTGT-3,U:5-CAACCCCAAAC2CACAACCATAA-3,M,U1.1.3PCRBiometra,1.21.2.1DNADNAQIAampDNABloodMiniKit,1ml,DNA-201.2.2DNAHerman[4],:DNA,,,PCR:50lDNA(2gDNA)NaOH3710min,10mmol/L30l3mol/L520l(pH5.0),,100l,5016hWizardDNAClean-UpSystemDNA,NaOH0.3mol/L,1015min,1/103mol/L3,-20DNA,70%,,DNA,-201.2.3p16DNA,PCR:4lDNA,1PCRBuffer,0.2mmol/LdNTP,0.2mol/L,HotStarTaq1.0U,50lPCR955min;:9530s,6030s,7230s,38725min8l,2.5%,SssIDNAMSP1.2.4SPSS13.02,P0.0522.1p1651,32DNAp16,62.7%,19p16,30p1611p16MSPMarker:DNA;P:;N:;M:;U:;M+:7720092161PracticalPreventiveMedicine,Feb.2009,Vol16,No.1©1994-2009ChinaAcademicJournalElectronicPublishingHouse.Allrightsreserved.(P0.05)11p162P()551155521140.3600.5492311981.0490.306TNM/1813/1460.7410.3893,,,,,p16,,,p16An[5]p1679.3%,[6],60%p16[7],,p16,,,DNA,DNA219ng/ml(),DNA[8,9]MSP,,DNA,,DNA,:,,[10]DNAMSPDNAp16,5132p16,62.7%,19,,p16,,p16,,,PCRp16,,[][1],.[J].,2001,24(8):450-4511[2]AllanJM,HardieLJ,BriggsJA,etal.GeneticalterationsinbronchialmucosaandplasmaDNAfromindividualsathighriskoflungcancer[J].IntJCancer,2001,91(3):359-3651[3]DasPM,SingalR.DNAmethylationandCancer[J].JClinOncol,2004,22(22):4632-46421[4]HermanJG,GraffJR,MyohanenS,etal.MethylationspecificPCR:anovelPCRassayformethylationstatusofCpGislands[J].ProcNatlAcadSciUSA,1996,93(18):9821-98261[5]AnQ,LiuY,GaoY,etal.Detectionofp16hypermethylationincir2culatingplasmaDNAofnon-smallcelllungcancerpati