Proteomic Analysis of Ovarian Cancer Cells Reveals Dynamic Processes of Protein Secretion and Shedding of Extra-Cellular Domains 英文参考文献.docVIP

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Proteomic Analysis of Ovarian Cancer Cells Reveals Dynamic Processes of Protein Secretion and Shedding of Extra-Cellular Domains 英文参考文献.doc

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Proteomic Analysis of Ovarian Cancer Cells Reveals Dynamic Processes of Protein Secretion and Shedding of Extra-Cellular Domains 英文参考文献

ProteomicAnalysisofOvarianCancerCellsReveals DynamicProcessesofProteinSecretionandSheddingof Extra-CellularDomains VitorM.Fac?a,AvivaP.Ventura,MathewP.Fitzgibbon,SandraR.Pereira-Fac?a, SharonJ.Pitteri,AnnE. Green,ReneeC.Ireton,QingZhang,HongWang,KathyC.O’Briant,CharlesW.Drescher,Miche`l Schummer,MartinW.McIntosh,BeatriceS.Knudsen,SamirM.Hanash* FredHutchinsonCancerResearchCenter,Seattle,Washington,UnitedStatesofAmerica Abstract Background: Elucidation of the repertoire of secreted and cell surface proteins of tumor cells is relevant to molecular diagnostics, tumor imaging and targeted therapies. We have characterized the cell surface proteome and the proteins releasedintotheextra-cellularmilieuofthreeovariancancercelllines,CaOV3,OVCAR3andES2andofovariantumorcells enrichedfromascitesfluid. MethodologyandFindings:Todifferentiateproteinsreleasedintothemediafromproteinconstituentsofmediautilized forculture,cellsweregrowninthepresenceof[13C]-labeledlysine.Abiotinylation-basedapproachwasusedtocapturecell surface associated proteins. Our general experimental strategy consisted of fractionation of proteins from individual compartmentsfollowedbyproteolyticdigestionandLC-MS/MSanalysis.Intotal,some6,400proteinswereidentifiedwith highconfidenceacrossallspecimensandfractions. ConclusionsandSignificance:Proteinprofilesofthecelllineshadsubstantialsimilaritytotheprofilesofhumanovarian cancercellsfromascitesfluidandincludedproteinmarkersknowntobeassociatedwithovariancancer.Proteomicanalysis indicatedextensivesheddingfromextra-cellulardomainsofproteinsexpressedonthecellsurface,andremarkablyhigh secretionratesforsomeproteins(nanogramspermillioncellsperhour).Cellsurfaceandsecretedproteinsidentifiedbyin- depthproteomicprofilingofovariancancercellsmayprovidenewtargetsfordiagnosisandtherapy. Citation:Fac?a VM,VenturaAP,FitzgibbonMP,Pereira-Fac?a SR,PitteriSJ,etal.(2008)ProteomicAnalysisofOvarianCancerCellsRevealsDynamicProcessesof ProteinSecretionandSheddingofExtra-CellularDomain