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1、Taoetal.JournalofExperimental&ClinicalCancerResearch(2015)34:139DOI10.1186/s13046-015-0253-3RESEARCHOpenAccessmiR-27b-3psuppressescellproliferationthroughtargetingreceptortyrosinekinaselikeorphanreceptor1ingastriccancer1†2†3†4441*4*JinqiuTao,XiaofeiZhi,Xiaoy
2、uZhang,MinFu,HaoHuang,YuFan,WenxianGuanandChenZouAbstractBackground:Thereceptortyrosinekinase-likeorphanreceptors(ROR)familycontainstheatypicalmemberROR1,whichplaysanoncogenicroleinseveralmalignanttumors.However,theclinicalpotentialsandunderlyingmechanismsof
3、ROR1ingastriccancerprogressionremainlargelyunknown.Inthisstudy,wevalidatedthemicroRNA-mediatedgenerepressionmechanisminvolvedintheroleofROR1.Methods:Bioinformaticprediction,luciferasereporterassay,quantitativereal-timePCR(qRT-PCR)andwesternblottingwereusedto
4、revealtheregulatoryrelationshipbetweenmiR-27b-3pandROR1.TheexpressionpatternsofmiR-27b-3pandROR1inhumangastriccancer(GC)specimensandcelllinesweredeterminedbymicroRNART-PCRandwesternblotting.Cellproliferation,colonyformationassayinsoftagarinvitroandtumorigeni
5、cityinvivowereperformedtoobservetheeffectsofdownregulationandupregulationmiR-27b-3pexpressiononGCcellphenotypes.Results:miR-27b-3psuppressedROR1expressionbybindingtothe3’UTRofROR1mRNAinGCcells.miR-27b-3pwassignificantlydownregulatedandreverselycorrelatedwith
6、ROR1proteinlevelsinclinicalsamples.AnalysisoftheclinicopathologicalsignificanceshowedthatmiR-27b-3pandROR1werecloselycorrelatedwithGCcharacteristics.EctopicmiR-27b-3pexpressionsuppressedcellproliferation,colonyformationinsoftagar,xenografttumorsofGCcells.Byc
7、ontrast,miR-27b-3pknockdownenhancedthesemalignantbehaviors.Ourstudiesfurtherrevealedthatthec-Src/STAT3signalingpathwaywasinvolvedinmiR-27b-3p-ROR1-mediatedcellproliferationregulation.Conclusions:TheseresultsshowthatmiR-27b-3psuppressesROR1expressionthroughth
8、ebindingsiteinthe3’UTRinhibitingthecellproliferation.ThesefindingsindicatethatmiR-27b-3pexertstumor-suppressiveeffectsinGCthroughthesuppressionofoncogeneROR1expressionandsuggestatherapeuticappli
