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  • 简介:Objective:Toinvestigatetheeffectsofanti-PML(promyelocyticleukemia)oranti-PML/RARa(promyelocyticleukemia/retionicacidreceptora)antisenseoligonucleotidesoncellgrowth,expressionofPML-RARamRNAandPML-RARa/PMLproteinlocationofNB4celllines.Methods:RT-PCRwasusedfordetectingPML-RARamRNAexpression,trypanblueexclusionforcellcount,methylcelloseassayforleukemiccolonyformingunitdetection,immuno-fluorescenceforPML-RARa/PMLproteinlocation.Results:Bothanti-PMLstartcodonregionantisence(STAS)andanti-PML-RARafusionregionantisence(FUAS)couldinhibitcellgrowthandtheformationofacutemyelocyticcolonyformingunitofcells(AML-CFU).Cellsbecomepartialdifferentiatedatdays5,beingmoreobviousinFUAS-treatedcellsthaninSTASones.DownregulationofPML-RARamRNAexpressionoccurredat24hoursinSTASandFUAS-treatedcellsandmaintainedforupto72hours.Immuno-fluorescenceanalysiswithanti-PMLmonoclonalantibodyshowedaremarkabledecreaseevencompletedisappearanceofmicrogranules.Theresidualgranulesbecameenlargedasdiscretedots(<10percell),similartonormalPODstructureinsomeSTAS-treatedcellsat24hours.At72hours,nearlyallthegranulesdisappeared.SimilarchangeswereobservedinFUAS-treatedcells.Conclusion:BothPMLandPML-RARaantisenceoligonucleotidescanspeciallyblocktheexpressionofPML-RARaatmRNAandproteinlevels.PMLproteinisimplicatedintheregulationsofcelldifferentiation.

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