c-Myc Regulates Self-Renewal in Bronchoalveolar Stem Cells 英文参考文献.docVIP

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c-Myc Regulates Self-Renewal in Bronchoalveolar Stem Cells 英文参考文献.doc

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c-Myc Regulates Self-Renewal in Bronchoalveolar Stem Cells 英文参考文献

c-MycRegulatesSelf-RenewalinBronchoalveolarStem Cells JieDong1,ShariSutor1,GuoqianJiang2,YajunCao1,YanW.Asmann2,DennisA.Wigle1* 1Division of General Thoracic Surgery, Department of Surgery, Mayo Clinic, Rochester, Minnesota, United States of America, 2Division of Biomedical Statistics and Informatics,DepartmentofHealthSciencesResearch,CollegeofMedicine,MayoClinic,Rochester,Minnesota,UnitedStatesofAmerica Abstract Background:Bronchoalveolarstemcells(BASCs)locatedinthebronchoalveolarductjunctionarethoughttoregenerate bothbronchiolarandalveolarepitheliumduringhomeostaticturnoverandinresponsetoinjury.Themechanismsdirecting self-renewalinBASCsarepoorlyunderstood. Methods:BASCs(Sca-1+,CD34+,CD312and,CD452)wereisolatedfromadultmouselungusingFACS,andtheircapacity for self-renewal and differentiation were demonstrated by immunostaining. A transcription factor network of 53 genes required for pluripotency in embryonic stem cells was assessed in BASCs, Kras-initiated lung tumor tissue, and lung organogenesis by real-time PCR. c-Myc was knocked down in BASCs by infection with c-Myc shRNA lentivirus. Comprehensive miRNA and mRNA profiling for BASCs was performed, and significant miRNAs and mRNAs potentially regulatedbyc-Mycwereidentified.Weexploredac-MycregulatorynetworkinBASCs usinganumberofstatisticaland computationalapproachesthroughtwodifferentstrategies;1)c-Myc/Maxbindingsiteswithinindividualgenepromoters, and2)miRNA-regulatedtargetgenes. Results: c-Mycexpressionwasupregulated inBASCsanddownregulatedoverthetime courseoflungorganogenesisin vivo.Thedepletionofc-MycinBASCsresultedindecreasedproliferationandcelldeath.MultiplemRNAsandmiRNAswere dynamically regulated in c-Myc depleted BASCs. Among a total of 250 dynamically regulated genes in c-Myc depleted BASCs, 57 genes were identified as potential targets of miRNAs through miRBase and TargetScan-based computational mapping.Afurther88geneswereidentifiedaspotentialdownstreamtargetsthroughtheirc-Mycbindingmotif. Co