GREB1 Functions as a Growth Promoter and Is Modulated by IL6STAT3 in Breast Cancer 英文参考文献.docVIP

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GREB1 Functions as a Growth Promoter and Is Modulated by IL6STAT3 in Breast Cancer 英文参考文献.doc

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GREB1 Functions as a Growth Promoter and Is Modulated by IL6STAT3 in Breast Cancer 英文参考文献

1DepartmentofMicrobiology,BiochemistryandImmunology,MorehouseSchoolofMedicine,Atlanta,Georgia,UnitedStatesofAmerica,2DepartmentofChemistry, Xavier University of Louisiana, New Orleans, Louisiana, United States of America, 3Department of Pathology, Department of Medicine, University of Miami School of Medicine,Miami,Florida,UnitedStatesofAmerica Abstract CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist. *E-mail:mliu@(ML);gwang@(GW);sguo@(SG) PLOSONE | 1 October2012 | Volume 7 | Issue 10 | e46410 GREB1FunctionsasaGrowthPromoter GREB1isInducedbyE2inER-positiveBreastCancerCell Line GREB1wasdetectedinMCF-7cellstreatedwithE2for24and 48hrs,whilenoproteinisdetectedintheER-negativeBT-549cellsor inMCF-7cellsgrowninestrogen-freeconditionsinWesternblotting assay(Figure1A).TreatmentwithICI182,780(ICI),anestrogen receptorantagonist,andsilencingtheGREB1genebysiRNAledto the loss of GREB1 protein expression. As shown in Figure 1B, GREB1proteinexpressionisreducedinMCF-7cellstreatedwith estrogenplusICI182,780comparedtothatobservedincellstreated withestrogenalone.Figure1CshowsthelossofdetectableGREB1 protein when GREB1 is knocked down by GREB1 siRNA (SiGREB1)at48hourscomparedtocontrol(CSiRNA),CSiRNA has no effect onE2-induced GREB1production. Corresponding densitometricanalysisofthebandsperformedwiththeImageQuant program(Bio-Rad)areshownbelowtheWesternblot.E2-induced GREB1mRNAlevelsweresimilarlyanalyzedasabove,withGREB1 mRNA levels correlating well with GREB1 protein expressions. UponE2stimulation,GREB1mRNAisnotableasearlyas24hours and lasts up to 48 hours as presented in a time course study in Figure1D.BT-549cellsexpressGREB1mRNAaslowasthecontrol. ICItreatment(Figure1E)andsilencingtheGREB1gene(Figure1F) bothsignificantlyreducedtheGREB1expressionatthetranscrip- tionallevel. GREB1mRNACorrelateswithERStatusinBreastCancer Patients,andPredictsPatientSurvivalandResponsesto TamoxifenTreatment GREB1isthemostsensitiveER-regulatedgeneinresponseto E2 stimulation in b