最新血浆miRNA表达谱与肺癌预后关系研究幻灯片.ppt

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1、血浆miRNA表达谱与肺癌预后关系研究(1)Inthenucleus,pri-miRNAsprocessedbyDroshaaspre-miRNAs;(2)pre-miRNAsexportedbyExportin-5intocytoplasm;(3)ProcessedbyDicerasmaturemiRNAs;(4)Perfectcomplementwith3’UTR:mRNAscleavage;in-perfectcomplement:translationalrepression.Nat.Rev.Genet.5(7):522-31,2004MicroRNAsandCan

2、cermiRNAasNovelImportantTumorSuppresserGenesandOncogenesThediscoveryofmicroRNAs(miRNAs)hasopenednewavenuesforcancerdiagnosisandpredictionoftreatmentresponseNatureReviewsCancer6:259-269,2006WhySerumBiomarkersareImportant?Easytoget(non-invasive)RepeatableInhighriskpopulation(cancerscreening)

3、ThediscoveryofstableserummiRNAsChenetal.2007CellRes.SolexaWholeGenomeSequencingmiRNAs(21-23nt)weremostabundantsequencesinserumRNAsSerummiRNAsinHealthyPeopleChenetal.2007CellRes.9092ExposureLungCancerClinicalOutcomeSerummiRNAexpressions–prognosis?ResearchQuestion:SerummiRNAandLungCancerSurv

4、ival30livecases34-68months30deadcases2-22months120patients123patientsCase-onlystudy(303cases)DiscoverySolexasequencingTrainingsetqRT-PCRValidationsetqRT-PCRRandomlyclassifiedSubjectsandAssaysStudyPopulation:Earlystage(I-IIIa)adenocarcinomaandsquamouscelllungcancerpatients,treatedwithbothop

5、erationandadjuvantchemotherapiesDiscoverysamples:“Longsurvival”patients:30livepatientssurvivalmorethan30months(34.6-61.8months)(StageI,10;stageII,10;stageIII,10)“Shortsurvival”group:30deadpatientssurvivallessthan25months(2.0-22.5months)(StageI,10;stageII,10;stageIII,10)Trainingandtestingsa

6、mplesets:243participantswereclassifiedas120fortrainingand123fortestingbycomputer-generatedrandomnumbers.qRT-PCRassaywasconductedtoquantifytheserumlevelsofmiRNAs.SerumRNAExtractionEfficiencyandtheReliabilityofqRT-PCRAssay0.1-1000fmolsynthesizedath-miR156amiRNAputinto250ulserum：R=0.9932miRNA

7、sdetectedin2repeatsamples：R=0.98CopiesofSolexaSequencingandqRT-PCRExpressionofthe11miRNAs(30LSvs30SS)miRNALongsurvivalgroupShortsurvivalgroupRatioP**NCopies*Mean±s.d.medianNCopies*Mean±s.d.medianmiR-486301630.658±0.3610.6693032051.129±0.5421.0650.050.0002miR-2