Capture of MicroRNA–Bound mRNAs Identifies the Tumor Suppressor miR-34a as a Regulator of Growth Factor Signaling 英文参考文献.docVIP

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Capture of MicroRNA–Bound mRNAs Identifies the Tumor Suppressor miR-34a as a Regulator of Growth Factor Signaling 英文参考文献.doc

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Capture of MicroRNA–Bound mRNAs Identifies the Tumor Suppressor miR-34a as a Regulator of Growth Factor Signaling 英文参考文献

CaptureofMicroRNA–BoundmRNAsIdentifiesthe TumorSuppressormiR-34aasaRegulatorofGrowth FactorSignaling AshishLal1,2,3.*,MarshallP.Thomas1,2.,GabrielAltschuler4.,FranciscoNavarro1,2.,ElizabethO’Day1,2 XiaoLingLi3,CarlaConcepcion5,Yoon-ChiHan5,JeromeThiery1,2,DanielleK.Rajani1,2 ,Aaron Deutsch1,2,OliverHofmann4,AndreaVentura5,WinstonHide4,JudyLieberman1,2* , 1ImmuneDiseaseInstitute,PrograminCellularandMolecularMedicine,Children’sHospitalBoston,Boston,Massachusetts,UnitedStatesofAmerica,2Departmentof Pediatrics,HarvardMedicalSchool,Boston,Massachusetts,UnitedStatesofAmerica,3GeneticsBranch,NationalCancerInstitute,NationalInstitutesofHealth,Bethesda, Maryland,UnitedStatesofAmerica,4DepartmentofBiostatistics,HarvardSchoolofPublicHealth,Boston,Massachusetts,UnitedStatesofAmerica,5Departmentof CancerBiologyandGenetics,MemorialSloanKetteringCancerCenter,NewYork,NewYork,UnitedStatesofAmerica Abstract AsimplebiochemicalmethodtoisolatemRNAspulleddownwithatransfected,biotinylatedmicroRNAwasusedtoidentify directtargetgenesofmiR-34a,atumorsuppressorgene.ThemethodreidentifiedmostoftheknownmiR-34aregulated genesexpressedinK562andHCT116cancercelllines.Transcriptsfor982geneswereenrichedinthepull-downwithmiR- 34ainbothcelllines.Despitethislargenumber,validationexperimentssuggestedthat,90%ofthegenesidentifiedin both cell lines can be directly regulated by miR-34a. Thus miR-34a is capable of regulating hundreds of genes. The transcripts pulled down with miR-34a were highly enriched for their roles in growth factor signaling and cell cycle progression. These genes form a dense network of interacting gene products that regulate multiple signal transduction pathways that orchestrate the proliferative response to external growth stimuli. Multiple candidate miR-34a–regulated genesparticipateinRAS-RAF-MAPKsignaling.EctopicmiR-34aexpressionreducedbasalERKandAKTphosphorylationand enhanced sensitivity to serum growth factor withdrawal, while cells genetically deficient in miR-34a were less sensi