137.full毕业论文.pdf

该【137.full毕业论文 】是由【巧姐】上传分享，文档一共【5】页，该文档可以免费在线阅读，需要了解更多关于【137.full毕业论文 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。CANCERGENOMICS&PROTEOMICS14:137-142(2017)doi:-eleratesInvasivenessofCancerCellsinRecurrentEpithelialOvarianCancerThroughRegulationofHomeoboxA9GUNOHCHONG1,HYO-SUNGJEON2,HYUNGSOOHAN3,JIWOONGSON4,YOONHEELEE1,DAEGYHONG1,HONGJUNPARK2,YOONSOONLEE1andYOUNGLAECHO11DepartmentofObstetricsandGynecology,SchoolofMedicine,KyungpookNationalUniversityHospital,Daegu,RepublicofKorea;.,Daegu,RepublicofKorea;3DepartmentofPhysiology,SchoolofMedicine,KyungpookNationalUniversityHospital,Daegu,RepublicofKorea;4DepartmentofInternalMedicine,KonyangUniversityHospital,Daejeon,:AlthoughmicroRNAs(miRNAs)ovariancancercellinvasivenessinrecurrentEOCbyaninareknowntoinfluencemessengerRNApost-:OverexpressionofmiR-controlandcontributetohumantumorigenesis,,miR-196bcanbeprimaryandrecurrentepithelialovariancancer(EOC).(EOC)isthemostlethalofallinvestigatewhethermiR-196bcouldregulatetheexpressiongynecologicalcancers,thefourthleadingcauseofcancer-oftheHomeoboxA9(HOXA9)gene,andthusaffecttherelateddeathsinwomenintheUnitedStates,:Microarrayswereusedtogeneratetheexpression(1).Ingeneral,lessthanhalf(45%)(2).,percentageofpatientsdiagnosedatanadvancedstage,whoassociatedgenes,:-andmiRNAs(includingmiR-196b)and18underexpressedtaxane--90%patients,mostresponderseventuallyHOXA9expressionwasinverselycorrelatedwithmiR--(miRNAs)aresmall(~22nucleotides)non--transcriptionallevels(3).ThesemoleculestypicallyCorrespondenceto:,(mRNA),NationalUniversityMedicalCenter,Daegu,Korea,807Hogukno,includingthatofgenesmediatingprocessesintumorigenesisBuk-gu,702-210Daegu,:+82532002681,suchasinflammation,cellcycleregulation,stressresponse,Fax:+82532002028,e-mail:******@-differentiation,apoptosis,,.,Daegu,Korea,Seongseogongdan-initiatedthroughspecificbase-pairinginteractionsbetweenthero11-gil,Dalseo-gu,Daegu,:+82532542505,Fax:+82532542507,e-mail:******@5’end("seed"region)ofthemiRNAandsiteswithinthecodinganduntranslatedregions(UTRs)ofmRNAs;targetsitesintheKeyWords:Epithelialovariancancer,recurrence,invasiveness,3’-UTRleadtomoreeffectivemRNAdestabilization(4).miR-196b,&PROTEOMICS14:137-142(2017),numerousmiRNAprofilingstudiesinEOChaveidentifiedmiRNAsassociatedwithchemotherapyresistanceanddiseaseprogression(5-8).However,,,,658humanmiRNAs,33miRNAswereoverexpressedand18miRNAswereunderexpressedinrecurrentEOC(9)..(Gibco?,Rockville,MD,USA)with10%fetalbovineserum(FBS)andantibiotics(penicillin100units/mlandstreptomycin100mg/ml).,whethermiR-196bmodulatesHOXA9expressionbybindingits3’-tumortissuespecimenswereobtainedfrom10KoreanpatientswithUTR,aluciferaseassaywasperformedusingpsiCHECK2plasmidEOC(primaryEOC,n=5;recurrentEOC,n=5)whounderwentcontainingtheHOXA93’-,035bpfragmentofHOXA9surgeryattheKyungpookNationalUniversityMedicalCenter,Daegu,3’-,psiCHECK2(Promega,Madison,WI,USA).Theforwardanizationcriteria,andthetumorhistotypewasdeterminedprimerwithaXhoIrestrictionsite(5’--3’)(5’-TwithrecurrentEOChadreceivedatleast6cyclesofplatinum-basedTGAGGTAAC-3’)wereusedtoamplifytheHOXA93’-UTRcombinationchemotherapy(paclitaxelpluscarboplatin).,-wellplatesinDMEMinliquidnitrogenandstoredat?80?%heat-inactivatedFBSandthentransfectedwerehistologicallyconfirmedbyhematoxylin--HOXA9constructscontainingthe3’-UTRofInstitutionalReviewBoardapprovedthestudyprotocol,andwrittenHOXA9,inthepresenceofamiR-(Ambion)usingtheEffectene(Qiagen,Hilden,Germany)(2μg)(Qiagen,Hilden,Germany)accordingtothemanufacturers’measuredusingaLumatLB953luminometer(EG&GBerthhold,,Germany),andtheresultswerenormalizedusingBioanalyzer(AgilentTechnologies,PaloAlto,CA,USA).(Affymetrix,SantaClara,CA,USA).Totaltriplicateusing48-wellmicrochemotaxischambers(,RNAwasthenhybridizedontoanAffymetrixGeneChip?miRNAInc.,Gaithersburg,MD,USA)with8-μmporemembranes(,Inc.,Gaithersburg,MD,USA)pre-coatedwith10μg/mlintensitieswerequantifiedandanalyzedusingtheGenechipMatrigel(BDBioscience).SK-OV-3cells(1×104)in50μlofoperatingsoftware(Affymetrix).Rawdatawerenormalizedbytheserum-freemediumwereseededintheupperchamber,andtheRobustMulti-arrayAverage(RMA)methodtoremovesystematiclowerchamberwasfilledwith26-27μlmediumcontaining10%,?C,cellsthatmigratedtotheusingaformulabasedonnormaldistributionandusesalinearlowersurfaceofthemembraneswerestainedwithaDiff-Quickkitmodeltoestimatevaluesonalog--(Sysmex,Kobe,Japan)-%crystalvioletandanalyzedbybetweenrecurrentandprimaryEOCtissues,-OV-(TRAVIGEN,Gaithersbug,MD,USA).3and293TwerepurchasedfromAmericanTypicalCultureCollection(?,HTB-77?,CRL-3216?,respectively,,statisticalManassas,VA,USA)andweremaintainedinDMEM(parisonsweremadeusingtheStudent’st-:MicroRNA196--OV-3cellinvasionbymiR-196btransfection.(A)Crystalviolet(%)stainingofinvadingcellsintheupperchamber.(B)-testanddatawereconsideredstatisticallydevelopmentandinadulttissues,aswellasincancer(14).significantwhenp<,HOXA9mightbeacandidatetargetofmiR--196bcouldregulatetheHOXA9genedirectly,’-,4miRNAsincludingmiR-containingtheputativemiR-196bbindingsequences,which551b,miR-19b,miR-196b,andmiR-3198,werewaspredictedtocontaintwobindingsites,position940-959significantlyoverexpressedinrecurrentEOCfromamongbpand1973-1995ofHOXA93’-,-196bforfurtherthenco-transfectedinto293TcellsalongwithmiR-196b,orstudiesbecauseitwassuggestedtobeapotentialmarkerscrambledmiRNA,andRenillaluciferaseactivitywasinpatientswithlungadenocarcinoma(10),Renillaactivityandoralcancer(11,12).However,theroleofmiR-196binwiththemiR-(p=).ThissuggeststhatmiR-analysisshowedthatmiR-196btranscriptionlevelwas196brepressesHOXA9transcriptionbydirectlybindingtosignificantlyhigherinrecurrentEOCthanthatinprimaryits3’--(averagescore)(averagescore)InductionofcancercellinvasionbymiR-(p=).determinewhethermiR-196bhasaneffectonovariancancerprogression,aninvasionassaywasperformedwithRepressionofHOXA9transcriptionbymiR--OV--196bwasadirecttargetgeneofmiR-196b,-OV-thetop10targetgenesincludingHOXA7,HOXC8,SMC3,,HOXA9,HMGA2,CTBS,ZMYND11,C1arf88,andoverexpressionofmiR-,HOXA9,invasivenessinSK-OV-3cells(Figure2),suggestingthatHOXA10,HOX11,andHOXA13areexpressedalongtheHOXA9mightbeinvolvedininhibitingaggressivebehaviorMüllerianductaxis(13).Especially,&PROTEOMICS14:137-142(2017)Discussionnormaltissuesinthecontextofbreastcancer(26).RecentreviewarticlesdemonstratedthatHOXgenesareessentialManagementofEOChasimprovedoverthelast20yearsdueregulatorsoftissueidentityanddrivetumorigenesisandbinationalprogressionthroughtheirinvolvementinregulatingprocesseschemotherapy,-binedwithsuchasdifferentiation,proliferation,adhesion,migration,andtaxanes(15).However,theoverallcurerateisonly30%.TheapoptosisinEOC(27).OurdatarevealedthatHOXA9down-issuesoftumorrecurrence,drugresistance,enhancedregulationledtoinvasivenessbymiR-,itinvasion,,,mesenchymal/stemcell-ellstumorsfromlargecohortsofEOCpatientswithdocumentededataareneededtofurtherelucidatethelargelyresponsibleforEOCrecurrence(16-18).SeveralmechanismsunderlyingHOXgenedysregulation,-200chavebeena