Repertoire of microRNAs in Epithelial Ovarian Cancer as Determined by Next Generation Sequencing of Small RNA cDNA Libraries 英文参考文献.docVIP

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Repertoire of microRNAs in Epithelial Ovarian Cancer as Determined by Next Generation Sequencing of Small RNA cDNA Libraries 英文参考文献.doc

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Repertoire of microRNAs in Epithelial Ovarian Cancer as Determined by Next Generation Sequencing of Small RNA cDNA Libraries 英文参考文献

RepertoireofmicroRNAsinEpithelialOvarianCanceras DeterminedbyNextGenerationSequencingofSmall RNAcDNALibraries StaciaK.Wyman2,RachaelK.Parkin2,PatrickS.Mitchell2,BrianR.Fritz2¤,KathyO’Briant3,AndrewK. Godwin4,NicoleUrban3,CharlesW.Drescher3,BeatriceS.Knudsen3,MuneeshTewari1,2* 1DivisionofClinicalResearch,FredHutchinsonCancerResearchCenter,Seattle,Washington,UnitedStatesofAmerica,2DivisionofHumanBiology,FredHutchinson Cancer Research Center, Seattle, Washington, United States of America, 3Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington,UnitedStatesofAmerica,4DepartmentofMedicalOncology,FoxChaseCancerCenter,Philadelphia,Pennsylvania,UnitedStatesofAmerica Abstract Background:MicroRNAs(miRNAs)aresmallregulatoryRNAsthatareimplicatedincancerpathogenesisandhaverecently shown promise as blood-based biomarkers for cancer detection. Epithelial ovarian cancer is a deadly disease for which improved outcomes could be achieved by successful early detection and enhanced understanding of molecular pathogenesisthatleadstoimprovedtherapies.Acriticalsteptowardthesegoalsistoestablishacomprehensiveviewof miRNAsexpressedinepithelialovariancancertissuesaswellasinnormalovariansurfaceepithelialcells. Methodology:Weusedmassivelyparallelpyrosequencing(i.e.,‘‘454sequencing’’)todiscoverandcharacterizenoveland knownmiRNAsexpressedinprimaryculturesofnormalhumanovariansurfaceepithelium(HOSE)andintissuefromthree ofthemostcommonhistotypesofovariancancer.DeepsequencingofsmallRNAcDNAlibrariesderivedfromnormalHOSE and ovarian cancer samples yielded a total of 738,710 high-quality sequence reads, generating comprehensive digital profilesofmiRNAexpression.Expressionprofilesfor498previouslyannotatedmiRNAsweredelineatedandwediscovered six novel miRNAs and 39 candidate miRNAs. A set of 124 miRNAs was differentially expressed in normal versus cancer samples and 38 miRNAs were differentially expressed across histologic subtypes of ovarian cancer. Taqman qRT-PC