Profiling B cell chronic lymphocytic leukemia by reverse phase protein array: Focus on apoptotic proteins

[摘要] Bcellchroniclymphocyticleukemia(CLL)ischaracterizedbytheaccumulationofBlymphocytesfromproliferativeactivityandapoptosisresistance.TheincreasedawarenessoftheimportanceofBcellreceptorsignalinginCLLhasraisednewopportunitiesfortargetedintervention.Herein,wedescribeastudyperformedwiththehigh‐throughputRPPA(reversephaseproteinarray)technique,whichallowedustosimultaneouslystudydifferentmoleculesinalargeseriesofpatients.WeanalyzedBlymphocytesfrom57patientswithCLLand11healthysubjects.Differentpathwayswereassessedforactivation/expressionofkeysignalingproteins.DataobtainedwerevalidatedbyWesternblottingandconfocalmicroscopy.TheRPPAinvestigationanditsvalidation,identified3seriesofproteins:1)moleculeswhoseexpressionlevelsreachedstatisticallysignificantdifferencesinCLLvs.healthycontrols(HSP70,Smac/DIABLO,cleavedPARP,andcleavedcaspase‐6);2)proteinswithapositivetrendofdifferenceinCLLvs.healthycontrols(HS1,γ‐tubulin,PKCα/β‐IIThr‐638/641,p38MAPKThr‐180/Tyr‐182,NF‐κBSer‐536,Bcl2Ser‐70andSrcTyr‐527);and3)moleculesdifferentiallyexpressedinpatientswithIGHVmutationsvs.thosewithoutmutations(ZAP70,PKC‐ζλ,Thr‐410/403,andCD45).Thisstudyidentifiedsomemolecules,particularlythoseinvolvedinapoptosiscontrol,whichcouldbeconsideredforfurtherstudiestodesignnewtherapeuticstrategiesinCLL...

[学科分类] 生理学

[关键词] signal transduction;lymphocyte;HSP70;Smac/DIABLO