Argonaute 2 Complexes Selectively Protect the Circulating MicroRNAs in Cell-Secreted Microvesicles 英文参考文献.docVIP

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Argonaute 2 Complexes Selectively Protect the Circulating MicroRNAs in Cell-Secreted Microvesicles 英文参考文献.doc

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Argonaute 2 Complexes Selectively Protect the Circulating MicroRNAs in Cell-Secreted Microvesicles 英文参考文献

Argonaute2ComplexesSelectivelyProtectthe CirculatingMicroRNAsinCell-SecretedMicrovesicles LiminLi1,DihanZhu1,LeiHuang1,JingZhang1,ZhenBian1,2,XiChen1,YuanLiu2,Chen-YuZhang1*, KeZen1* 1JiangsuEngineeringResearchCenterforMicroRNABiologyandBiotechnology,StateKeyLaboratoryofPharmaceuticalBiotechnology,SchoolofLifeSciences,Nanjing University,Nanjing,Jiangsu,China,2CMBP,DepartmentofBiology,GeorgiaStateUniversity,Atlanta,Georgia,UnitedStatesofAmerica Abstract Cell-secreted miRNAs are highly stable and can serve as biomarkers for various diseases and signaling molecules in intercellularcommunication.ThemechanismunderlyingthestabilityofcirculatingmiRNAs,however,remainsincompletely understood.HereweshowthatArgonaute2(Ago2)complexesandmicrovesicles(MVs)providespecificandnon-specific protection for miRNA in cell-secreted MVs, respectively. First, the resistance of MV-encapsulated miRNAs to RNaseA was bothdependedonintactvesicularstructureofMVsandprotease-sensitive.Second,animmunoprecipitationassayusinga probecomplementary tohumanmiR-16,amiRNAprimarily located intheMVsandshowedastrong,protease-sensitive resistance to RNaseA, identified Ago2 as a major miR-16-associated protein. Compared with protein-free miR-16, Ago2- associatedmiR-16wasresistanttoRNaseAinadose-andtime-dependentfashion.Third,whenthemiR-16/Ago2complex wasdisruptedbytrypaflavine,theresistanceofmiR-16toRNaseAwasdecreased.Incontrast,whentheassociationofmiR- 16withtheAgo2complexeswasincreasedduringcellapoptosis,althoughthetotalamountofmiR-16andAgo2remained unchanged,theresistanceofmiR-16toRNaseAintheMVswasenhanced.AsimilarcorrelationbetweentheincreaseofmiR- 223/Ago2 association and the resistance of miR-223 against RNaseA was observed during all trans retinoic acid (ATRA)- inducedcelldifferentiationofpromyelocyticHL60cells.Inconclusion,theassociationofmiRNAswithAgo2complexes,an eventthatislinkedtocellfunctionalstatus,playsacriticalroleinstabilizingthecirculatingmiRNAsincell-secretedMVs. Citation: Li L, Zhu D, Hu