A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling 英文参考文献.docVIP

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A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling 英文参考文献.doc

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A Dual-Color Fluorescence-Based Platform to Identify Selective Inhibitors of Akt Signaling 英文参考文献

ADual-ColorFluorescence-BasedPlatformtoIdentify SelectiveInhibitorsofAktSignaling Aranzazu′ Rosado.¤,FabianZanella.,BeatrizGarcia,AmancioCarnero,WolfgangLink* ExperimentalTherapeuticsProgram,CentroNacionaldeInvestigacionesOncologicas(CNIO),Madrid,Spain Abstract Background:InhibitionofAktsignalingisconsideredoneofthemostpromisingtherapeuticstrategiesformanycancers. However, rational target-orientated approaches to cell based drug screens for anti-cancer agents have historically been compromisedbythenotoriousabsenceofsuitablecontrolcells. Methodology/Principal Findings: In order to address this fundamental problem, we have developed BaFiso, a live-cell screeningplatformtoidentifyspecificinhibitorsofthispathway.BaFisoreliesontheco-cultureofisogeniccelllinesthat havebeenengineeredtosustaininterleukin-3independentsurvival oftheparental Ba/F3 cells,andthat areindividually tagged with different fluorescent proteins. Whilst in the first of these two lines cell survival in the absence of IL-3 is dependent on the expression of activated Akt, the cells expressing constitutively-activated Stat5 signaling display IL-3 independentgrowthandsurvivalinanAkt-independentmanner.Smallmoleculescanthenbescreenedintheselinesto identifyinhibitorsthatrescueIL-3dependence. Conclusions/Significance: BaFiso measures differential cell survival using multiparametric live cell imaging and permits selectiveinhibitorsofAktsignalingtobeidentified.BaFisoisaplatformtechnologysuitablefortheidentificationofsmall moleculeinhibitorsofIL-3mediatedsurvivalsignaling. Citation:RosadoA,ZanellaF,GarciaB,CarneroA,LinkW(2008)ADual-ColorFluorescence-BasedPlatformtoIdentifySelectiveInhibitorsofAktSignaling.PLoS ONE3(3):e1823.doi:10.1371/journal.pone.0001823 Editor:MarkR.Cookson,NationalInstitutesofHealth,UnitedStatesofAmerica ReceivedNovember20,2007;AcceptedFebruary4,2008;PublishedMarch19,2008 Copyright: ? 2008 Rosado et al. This is an open-access article distributed under the terms of the Creative Commons At