Benzimidazoles Downregulate Mdm2 and MdmX and Activate p53 in MdmX Overexpressing Tumor Cells Zuzana Mrkvová.pdf

该【Benzimidazoles Downregulate Mdm2 and MdmX and Activate p53 in MdmX Overexpressing Tumor Cells Zuzana Mrkvová 】是由【dt83088549】上传分享，文档一共【14】页，该文档可以免费在线阅读，需要了解更多关于【Benzimidazoles Downregulate Mdm2 and MdmX and Activate p53 in MdmX Overexpressing Tumor Cells Zuzana Mrkvová 】的内容，可以使用淘豆网的站内搜索功能，选择自己适合的文档，以下文字是截取该文章内的部分文字，如需要获得完整电子版，请下载此文档到您的设备，方便您编辑和打印。:..moleculesArticleBenzimidazolesDownregulateMdm2andMdmXandActivatep53inMdmXOverexpressingTumorCellsZuzanaMrkvová1,StjepanUldrijan1,2,AntonioPombinho3,PetrBart?unˇek3andIvaSlaninová1,*1DepartmentofBiology,FacultyofMedicine,MasarykUniversity,Kamenice5,BuildingA6,62500Brno,CzechRepublic;zuzana.******@(.);******@(.)2InternationalClinicalResearchCenter,’sUniversityHospital,65691Brno,CzechRepublic3CZ-OPENSCREEN:NationalInfrastructureforChemicalBiology,icsASCR,.,14220Prague,CzechRepublic;antonio.******@(.);******@(.)*Correspondence:******@;Tel.:+42-05-4949-6985AcademicEditors:,Karel?mejkalandElkeHeiss



Received:10May2019;Accepted:5June2019;Published:7June2019Abstract:Tumorsuppressorp53ismutatedinabout50%-typep53,,monlyusedbenzimidazoleanthelmintics,,indicatingactivationofthep53–p21pathway,,.,G2/,-:benzimidazoles;drugrepurposing;Mdm2;MdmX;melanoma;%ofhumanmalignantneoplasmscarrymutationsinthep53gene(TP53).Additionally,itsfunctionisabrogatedbytheoverexpressionofMdm2andMdmXinasignificantproportionoftumorsretainingwild-typep53[1,2].Mdm2servesasanE3ubiquitinligasethattargetsp53forproteasome-mediateddegradation[3].MdmXandMdm2candimerizeviatheirRING?ngerdomains,andthisinteractionregulatescellularlevelsofp53andMdmX,aswellasMdm2[1,4].Ampli?cationoroverexpressionofMdm2andMdmXoccurinawiderangeoftumors,includingmalignantmelanoma[5].Notably,MdmXoverexpressionwasdetectedinapproximately65%ofmelanomasinstagesI–IV[6].Giventhatmutationsofp53areursinmelanoma,,,24,2152;doi::..MoleculesMolecules20192019,24,,215224,xFORPEERREVIEW2of142of14Malignantmelanomaisanaggressiveformofskincancerwithapproximately133,000newlyThemorethediseasedevelops,thelowerthe?ve-es:Startingat100%?nishingat7–19%duringtheexpansionofdistantmetastases[morethediseasedevelops,thelowerthefive-es:Startingat100%during7].Currenttherapeuticapproachesincludesurgicalresection,chemotherapybasedonalkylatingagentisolatedhorizontalgrowthandfinishingat7–19%duringtheexpansionofdistantmetastases[7].dacarbazine,photodynamictherapy,andimmunotherapyusinginterferonCurrenttherapeuticapproachesincludesurgicalresection,chemotherapybasedonalkylatingagent-2b,interleukin-2,andanti-PD-,newtherapeutictargetshavebeendacarbazine,photodynamictherapy,andimmunotherapyusinginterferonα-2b,interleukin-2,andanti-PD-,newtherapeutictargetshavehangesa,suchassmallmoleculeinhibitorsorantibodiesthataapplicationofthetargetedtherapy,suchassmallmoleculeinhibitorsorantibodiesthataffectthe%,%-mutantinhibitors,wereapprovedbytheFDAforthetreatmentofmelanomascarryingthedabrafenib,selectiveBRAF-mutantinhibitors,wereapprovedbytheFDAforthetreatmentofmonactivatingmutationBRAFV600E[8].Despitetheseadvancesintherapy,itseV600EciencyismonactivatingmutationBRAF[8].Despitetheseadvancesinfrequentlydecreasedduetothedevelopmentofdrugresistanceandnewtreatmentstrategiesneedtherapy,ectivecancertherapyisdrugrepositioning(drugrepurposing),.,redirectingexistingdrugstonewindications[effectivecancertherapyisdrugrepositioning(drugrepurposing),.,redirectingexistingdrugsto9].Whencomparedtotheapprovalprocessofnewdrugs,priselowerexpensesnewindications[9].paredtotheapprovalprocessofnewdrugs,-throughputscreeningofmorethan2000pharmacologicallyrelevantcompoundsinordertoidentifyagentswiththepotentialtoincreasethep53transcriptionalactivityWeperformedhigh-,poundsinordertoidentifyagentswiththepotentialtoincreasethep53transcriptionalactivitypounds,albendazole(ABZ,Figure1A)andfenbendazole(FBZ,Figure1B),weselectedtwopounds,albendazole(ABZ,Figure1A)andfenbendazole(FBZ,Figure1B),thesameTheprimarymechanismoftheiractionisthedisruptionofthecellularmicrotubularsystem,thesamemechanismbywhichtwowidelyusedgroupsofcytostatics,theVincaalkaloidsandtaxanes,,theVincaalkaloidsandtaxanes,-bindingsiteintubulin[Benzimidazolesbindtothecolchicine-bindingsiteintubulin[10].Sincetheybindtotheirown10].Sincetheybindtotheirowndistinctbindingsites,etheresistancetothecytostaticsmentionedabove[distinctbindingsites,etheresistancetothecytostaticsmentionedabove[11].ABZ11].ABZcouldsuppressthegrowthofpaclitaxel-resistantleukemia(CEMcouldsuppressthegrowthofpaclitaxel-resistantleukemia(CEM/dEpoB300)andovarian/dEpoB300)andovarian(1A9PTX22)tumorcells[(1A9PTX22)tumorcells[12,13].Theresponseofpaclitaxel-resistantcellstoABZwasevenenhanced12,13].Theresponseofpaclitaxel-paredtotheparentalleukemiacells[12].comparedtotheparentalleukemiacells[12].(Chemicalstructuresof(AA)albendazoleand()albendazoleand(BB)fenbendazole.)[andwasintroducedtohumanmedicinein1982[14].ThemechanismofABZactionincludesnotonly14].ThemechanismofABZactionincludesnotonlythemicrotubuleinhibitionbutalsotheblockageofglucoseuptakeandgenerationofROS,leadingtothemicrotubuleinhibitionbutalsotheblockageofglucoseuptakeandgenerationofROS,leadingtolowcellularlevelsofATP[lowcellularlevelsofATP[15]andDNAdamage[16,17],]andDNAdamage[16,17],ectsincancercellarcinoma[arcinoma[16],acutelymphoblasticleukemia[12],16],acutelymphoblasticleukemia[12],hepatocellularcarcinoma[hepatocellularcarcinoma[18],andcolorectalcarcinoma[19].RegardingFBZ,],andcolorectalcarcinoma[19].RegardingFBZ,-binedwithagrowthinhibitionoflymphomaxenograftedintoSCIDmicecausedbyafenbendazole-containinghighdoseofvitamins[20].binedwithahighdoseofvitamins[20].Inthelightofthe?ndingsthatmalignantmelanomabelongstotumorsretainingwild-typep53butitsfunctionislimitedbytheoverproductionofitsnegativeregulatorsMdm2andMdmX,:..Molecules2019,24,21523of14Molecules2019,24,,Inthelightofthefindingsthatmalignantmelanomabelongstotumorsretainingwild-typep53wedemonstratethattwoclinicallyapprovedanthelminticsABZandFBZincreasep53activityincancerbutitsfunctionislimitedbytheoverproductionofitsnegativeregulatorsMdm2andMdmX,,,weactivityofABZandFBZ,thedisruptionofmicrotubules,cellcyclearrestattheG2demonstratethattwoclinicallyapprovedanthelminticsABZandFBZincreasep53activityincancer/Mcheckpoint,,weconfirmedtheantiproliferativeactivityofABZandFBZ,thedisruptionofmicrotubules,cellcyclearrestattheG2/,ectofsmallmoleculedrugsonp53transcriptionalactivityinmelanoma,weestablishedA375celllinecarryingp53activityluciferasereporterconstruct(A375-p53-ells)andperformedluciferasereporterassays(Figure2A).pounds,thestablyInordertoinvestigatetheeffectofsmallmoleculedrugsonp53transcriptionalactivityinmelanoma,weestablishedA375celllinecarryingp53activityluciferasereporterconstruct(A375-binationwiththep53-ells)andperformedluciferasereporterassays(Figure2A).,dose-pounds,,ectedthep53activityinA375-p53-ellsarepresentedinthesupplement(TableS1,TableS2,andFigureS1).Basedonthescreening,wechosebenzimidazoledose-responsedata,poundsthataffectedthep53activityinA375-p53-LucderivativesABZandFBZforamoredetailedstudy(Figure2B).A375-p53-ellsweretreatedcellsarepresentedinthesupplement(TableS1,TableS2,andFigureS1).Basedonthescreening,wewithABZ(2chosebenzimidazolederivativesABZandFBZforamoredetailedstudy(Figure2B).A375-p53-LucM)andFBZ(1M)?edCDKinhibitorsaspotentnon-genotoxicactivatorsofp53inmelanomacells[cellsweretreatedwithABZ(2μM)andFBZ(1μM),22].Forthisreason,CDKinhibitordinaciclib(DINA;40nM)-genotoxicactivatorsofp53inmelanomacells[21,22].Forthisreason,CDKinhibitordinaciclib(DINA;40nM)%,ABZto247%,andFBZto504%paredtosolventresultsshowedthatDINAincreasedp53activityto865%,ABZto247%,andFBZto504%when(DMSO)-treatedcontrolcells(Figure2C).comparedtosolvent(DMSO)-treatedcontrolcells(Figure2C)..(construct.(A)pounds.(A)pounds.(B)HTSrevealedalbendazole(A