st KAPAK RESMİ KONACAK - hucreolumu.org · Through Affecting STAT Signalling Differently In...

1 1stInternationalCellDeathResearchCongress-Turkey

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KAPAKRESMİKONACAK


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1stInternationalCellDeathResearchCongress-Turkey.AbstractBook(2ndCongressofCellDeathResearchSociety-Turkey)2.HücreÖlümüAraştırmaDerneğiKongresi-Türkiye.ÖzetKitabı

Editors:Prof.Dr.A.SemraKOÇTURK,PhDDokuzEylülUniversity,FacultyofMedicine,DepartmentofBiochemistry,Izmir,Turkey.Prof.H.SedaVATANSEVER,MD,PhDCelalBayarUniversity,FacultyofMedicine,DepartmentofHistology-Embryology,Manisa,Turkey.

CoverPageDesign:DalyaTourismCompany-İzmir-TurkeyPublisher:CellDeathResearchSociety(HücreÖlümüAraştırmaDerneği)-İzmir-TurkeyLanguage:EnglishISBN-13:978-605-63544-4-1FirstEdition(electronic):May2016-İzmir,TurkeyAllRightsreservedbyCellDeathResearchSocietyofTurkey.Thisbookmaynotbereproducedinwholeorpartwithoutpermission.Makingordistributingelectroniccopiesofthisbookpostitutescopyrigtinfringementandcouldsubjecttheinfiringertocriminalandcivilliability.


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DearColleagues,OnbehalfoftheCellDeathResearchSocietyofTurkey(HÖAD,HücreÖlümüAraştırmaDerneği)itisagreathonorandpleasuretoinviteyoutothe1stInternationalCellDeathResearchCongress,whichwillbeheldon04-07ofMay2016inIzmir,TURKEY.Thecongress is scientific refereed.Themain topicsof thecongresswillbe the focusingon the importanceandroleofdifferenttypesofcelldeathinoxidativestress,immunity,stemcelltherapies,genomesignatures,therapeuticapproachesandpolyphenolsasanti-canceragentindiseases.Thecongresswill featureplenary,keyandshort lecturesbesidesposterpresentations.Ourmaingoal is togatherthescientistsfromuniversities,researchcentersfromallovertheworldandoffertoallagreatandattractivecongressjoiningdifferentknowledgetogetheroncelldeath.Anumberoftheinternationallydistinguishedspeakerswiththeexpertiseinthefieldsareinvited.Theywillintroduce the cutting-edge research and the future perspectives relevant to the subjects covered in thepresentmeeting.Besides,oralandposterpresentationsareincludedformorescientificdiscussions.Twoyearsago,thefirstcongressofthesocietywasinternationalparticipatingandoverthan300researchersfrom all over world mostly US and Europe including the 46 speakers were attended. Therefore, we areexpectingasimilarorhigherparticipationonthedateofMay2016,whichisverynicetimeoftheseasoninİzmir,Turkey.Congress will be held in Dokuz Eylul University School ofMedicine in Izmir - one of the preeminent andbiggestuniversities inTurkey. Izmirregion itself,neartheAegeanSea, is famousfor itsrichculture,historyandoutstandingscenery.We would like to take this opportunity to invite all researchers and accompanying parties to attend thismeetingandtoenjoytherichscientificprogramandthebeautyofTurkeyandTurkishhospitality.Sincerelyyours,OnbehalfoftheOrganizingCommittee,Iwouldappreciateyourinterestandhopetowelcomeyouinİzmir.ProfessorSemraKOÇTÜRK,Ph.DPresident,CellDeathResearch Society-Turkey


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COMMITTEES

OrganizationCommittee:

CellDeathResearchSocietyofTurkey

SemraKOCTURK(PresidentofCDRS–Turkey)

KemalS.KORKMAZ(VicePresidentofCDRS-Turkey)

H.SedaVATANSEVER(GeneralSecretaryofCDRS–Turkey)

ZekiyeALTUN(TreasurerofCDRS-Turkey)

PınarAKAN(MemberofCDRSCommittee-Turkey)

DevrimGOZUACIK(MemberofOrganizationCommittee)

AytenNALBANT(MemberofOrganizationCommittee)

MauroPIACENTINI(MemberofOrganizationCommittee)

ScientificSecretary:

H.SedaVATANSEVER

SocialCommittee:

YunusAKKOC

JohannaAGGREY-FYNN

UgurBORA

FulyaCAGLAR

BengisuGELMEZ

SecilERBIL

PınarERCETIN

HilalKABADAYI

RemziyeKENDIRCI

TunaONAL

GunesOZEN

ErsinOZTURK

IrodaSAYDULLAYEVA

EfeSERINAN

BelginSERT


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ScientificAdvisoryBoard:AliUgurURAL(BayındırHospitalsGroup-Turkey)AlpCan(AnkaraUniversity-Turkey)AndrasNAGY(Lunenfeld-TanenbaumResearchInstitute-Canada)AyhanBILIR(İstanbulUniversity-Turkey)AytenNALBANT(IzmirInstituteofTechnology-Turkey)BharatBAGGARWAL(M.D.AndersonCancerCenter-USA)BorisZHIVOTOVSKY(KarolinskaInstitute-Sweden)BulentOZPOLAT(M.D.AndersonCancerCenter-USA)CerenKORKMAZ(EgeUniversity-Turkey)DevrimGOZUACIK(SabancıUniversity-Turkey)DoganYUCEL(PresidentofTurkishBiochemicalSociety-Turkey)ElifDamlaARISAN(IstanbulKültürUniversity-Turkey)EmelEKSIOGLU(MarmaraUniversity-Turkey)EnginULUKAYA(PresidentofMolecularCancerResearchSociety-Turkey)ErdalKARAOZ(PresidentofStemCellandCellularMedicineSociety-Turkey)EsraERDAL(DokuzEylülUniversity-Turkey)FahriSAATCIOGLU(UniversityofOslo-Norway)FerhanSAGIN(Co-PresidentofTurkishBiochemicalSociety-Turkey)FrancescoCECCONI(DanishCancerSocietyResearchCenter-Italy/Denmark)GabrielLOPEZ-BERESTEIN(M.D.AndersonCancerCenter-USA)GianLuigiRUSSO(InstituteofFoodSciencesAvellino-Italy)GulinnazERCAN(EgeUniversity-Turkey)GunnurDENIZ(PresidentofTurkishImmunologySociety-Turkey)H.SedaVATANSEVER(CelalBayarUniversity-Turkey)HakanAKBULUT(AnkaraUniversity-Turkey)HavalSHIRWAN(UniversityofCaliforniaatSantaBarbara-USA)HilalKOCDOR(PresidentofTurkishBiochemicalSocietyIzmirBranch-Turkey)IanDRANSFIELD(UniversityofEdinburgh-UK)IlknurKOZANOGLU(BaşkentUniversity-Turkey)IsilAKSANKURNAZ(GebzeTechnicalUniversity-Turkey)KemalS.KORKMAZ(EgeUniversity-Turkey)MauroPIACENTINI(UniversityofRome-Italy)MuratOZGOREN(VicePresidentofDokuzEylülUniversity-Turkey)NesrinOZOREN(PresidentofMolecularBiologyandGeneticSociety-Turkey)NurOLGUN(ManagerofDEUInstituteofOncology-Turkey)PeterVANDENABEELE(PresidentofEuropeanCellDeathOrganization-Belgium)PınarAKAN(DokuzEylülUniversity-Turkey)SefikALKAN(AlbaTherapeutics-USA)SemraKOCTURK(DokuzEylülUniversity-Turkey)SerifSENTURK(DokuzEylülUniversity-Turkey)SevdaMUFTUOGLU(HacettepeUniversity-Turkey)SevimAYDIN(AnkaraUniversity-Turkey)TuncayDEMIRYUREK(GaziantepUniversity-Turkey)TurgutULUTIN(PresidentofMedicalBiologyandGeneticSociety-Turkey)UfukGUNDUZ(MiddleEastTechnicalUniversity-Turkey)UmitZEYBEK(PresidentofTurkishMolecularMedicineSociety)VojoDERETIC(UniversityofNewMexico-USA)VolkanSEYRANTEPE(IzmirInstituteofTechnology-Turkey)ZekiyeALTUN(DokuzEylülUniversity-Turkey


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TopicsofTheCongress:

Session1: CellularMechanismsSession2: OxidativestressandcelldeathSession3: CancerandcelldeathSession4: Stemcell,PluripotencyandcelldeathSession5: PolyphenolsincelldeathSession6: ImmunityandcelldeathSession7: Therapeuticapproachestocelldeath


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ScientificProgramme

1stINTERNATIONALCELLDEATHRESEARCHCONGRESS-TURKEY(2ndCONGRESSOFCELLDEATHRESEARCHSOCIETY-TURKEY)

04-07MAY2016TURKEY-IZMIR

MAY4th2016(WEDNESDAY)

08:30-17:30 Registration

09:00-10:30

OpeningCeremony:*SemraKOCTURK(PresidentofCDRS-Turkey)*PinarTUNCEL(ViceDeanofDEUSchoolofMedicine)*MuratOZGOREN(ChairmanofDEPARKExecutiveBoard)*MehmetFUZUN(RectorofDEU)*MusicRecital-REYQuartet,Performedby;ZeynepSimheACUNAZ(Violin)CeydaOZDEMIR(Violin)IrisICELLIOGLU(Viola)BerkKAVAK(Violoncello)*Visualshow:AQUAgraphs.LightsWrittenonWater,PresentedbyAlpCAN

10:30-11:00 CoffeeBreak

11:00-12:00

OpeningLectureChair:MauroPIACENTINIRoleOfRIPKinasesInRegulatingCellDeathandSurvivalInVitroandInVivo,PeterVANDENABEELE(PresidentofECDO)

12:00-13:00 LUNCH

13:00-13:30 CompanyPresentation-FLUIDIGM:ThePolarisSystem:IntegratingCellandMolecularAnalysis,GregoryGONZALEZ

13:30-15:15 Session1:CellularMechanismsChairs:PeterVANDENABEELE,DevrimGOZUACIK

13:30-14:15 Type2Transglutaminase:AKeyRegulatorOfProteostasisUnderCellularStressfulConditions,MauroPIACENTINI(KeynoteLecture)

14:15-15:00 AutophagyAtTheIntersectionBetweenCellSurvivalandCellDeath:RolesInInflammationandLysosomalHomeostasis,VojoDERETIC(KeynoteLecture)

15:00-15:30 NovelRegulatorsOfAutophagy,DevrimGOZUACIK15:30-16:00 CoffeeBreak

16:00-17:00

OpenDiscussionwithEditorialMembersofJournalsModerator:KemalS.KorkmazMauroPIACENTINIBorisZHIVOTOVSKYFrancescoCECCONIPeterVANDANABEELEYahyaLALELIEkremGUREL


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17:00-19:00

OpenDiscussionProblems,SuggestionsforSolutionsinHealthResearchinTurkey(SessionwillbeheldinTurkish)Moderator:DevrimGOZUACIKSevimAYDIN-TÜBİTAK-SBAGBaşkanıMuratOZGOREN-DEPARKYönetimKuruluBaşkanıEsraERDAL-IBGİzmirMüdürYardımcısı

19:30-22:30 OPENINGCOCKTAIL–AQUAMARIN

MAY5th2016(THURSDAY)

09:00-10:45 Session2:OxidativeStressandCellDeathChairs:FerhanSAGIN,HilalKOCDOR

09:00-09:45 MitochondrialSubstrates:AToolToCombatCancer,BorisZHIVOTOVSKY(Keynotelecture)

09:45-10:15 AMBRA1NegativeControlAtTheCrossroadAmongAutophagy,CellProliferationandCellDeath,FrancescoCECCONI

10:15-10:45 InVitroandInVivoCharacterizationOfCellSurvivalGenesUsingDestabilizedCas9,SerifSENTURK10:45-11:15 CoffeeBreak

11:15-12:15

OralPresentationsSession-2

ConferenceHallChair:SemraKOCTURK

Session-1Classroom-B(Downstairs)

Chair:SaimeBATIRELOP-1BilgeDebelecBUTUNER OP-1MimouneBEREHABOP-2MehmetErayALCIGIR OP-2OzlemORALOP-3PinarERKEKOGLU OP-3GulceSariKAPLANOP-4OzgeCAGLAR OP-4UlviAHMADOV OP-5ElginTurkozULUER

12:15-13:30 LUNCH

13:30-15:15 Session3:CancerandCellDeathChairs:FahriSaatcioglu,KemalS.Korkmaz

13:30-14:15 STAMPingProliferationandCellDeathInProstateCancer,FahriSAATCIOGLU(KeynoteLecture)

14:15-14:45 ETSTranscriptionFactors-CriticalRegulatorsOfBrainTumorInitiatingCellProliferationvsNeurodegeneration?,IsilAKSANKURNAZ

14:45-15:15 DiminishedCyclinDependentKinaseActivityandmTORAreCriticalInTheCellDeathDecisionThroughAffectingSTATSignallingDifferentlyInProstateCancerCells,ElifDamlaARISAN


15:45-17:35 Session4:StemCell,PluripotencyandCellDeathChairs:AlpCAN,H.SedaVATANSEVER

15:45-16:15 TheBrightandTheDarkSidesOfReprogrammingToPluripotency,AndrasNAGY(KeynoteLecture)16:15-16:45 StemCellTherapyApproachesToIschemicCardiomyopathy,AlpCAN

16:45-17:15


ConferenceHallChair:KemalS.KORKMAZ

Session-4Classroom-B(Downstairs)Chair:GulinnazERCAN

OP-1BetulKARADEMIR OP-1FeyzanOzdalKURTOP-2TunaONAL OP-2HilalKABADAYIOP-3AysunEKINCI OP-3RemziyeKENDIRCI

17:15-18:30 POSTERSESSION-Refreshment18:30-19:30 GENERALASSEMBLYOFCELLDEATHRESEARCHSOCIETYOFTURKEY


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MAY6th2016(FRIDAY)

09:00-10:45 Session5:PolyphenolsinCellDeathChairs:BulentOZPOLAT,SemraKOCTURK

09:00-09:45 TargetingInflammatoryandApoptoticPathwaysByAgentsDesignedByMotherNatureForPreventionandTreatmentOfCancer,BharatAGGARWAL(Keynotelecture)

09:45-10:15 MolecularMechanismsOfFlavonoidQuercetinInEnhancingApoptosisInChronicLymphocyticLeukemia,GianLuigiRUSSO

10:15-10:45 DevelopmentOfNovelTargetedTherapiesForSolidTumors,BulentOZPOLAT10:45-11:15 CoffeeBreak

11:15-12:15


ConferenceHallChair:ZekiyeS.ALTUN

Session-3-4-5-6Classroom-B(Downstairs)Chair:BetulKARADEMIR

OP-1MehmetFatihSEYHAN OP-1SerapCELEBIOP-2AyşeMineYILMAZ OP-2MehmetKadirERDOGANOP-3AbdullahTuncayDEMIRYUREK OP-3LeventELMASOP-4GunesOZEN OP-4SaimeBATIRELOP-5IbrahimBOZGEYIK OP-5CeylanHEPOKUROP-6AyferKARLITEPE

12:15-13:30 LUNCH

13:30-15:15 Session6:ImmunityandCellDeathChair:VolkanSEYRANTEPE

13:30-14:15 MerReceptorTyrosineKinaseandMacrophagePhagocytosisOfApoptoticCells,IanDRANSFIELD14:15-14:45 UnequalCellDeathInTheDifferentiationOfTHelperCellSubsets,AytenNALBANT

14:45-15:15


ConferenceHallChair:IsilKURNAZ

Session-7Classroom-B(Downstairs)Chair:TuncayDEMIRYUREK

OP-1AytenNALBANT OP-1HaticeMehtapKUTLUOP-2FerdiyeTANER OP-2ZekiyeS.ALTUNOP-3SeminayGULER OP-3MelikeOZGUL


15:45-17:30 Session7:TherapeuticApproachestoCellDeathChairs:HavalSHIRWAN,PinarAKAN

15:45-16:30 Monocytes:KillersorSaviors,GabrielLOPEZ-BERESTEIN

16:30-17:00 ApoptosisAsAPowerfulMeansOfImmuneModulationForTheTreatmentOfType1Diabetes,HavalSHIRWAN

17:00-17:30 AbnormalBrainGangliosideAccumulationTriggersApoptosisInEarlyOnsetTay-SachsDiseaseMouseModel,VolkanSEYRANTEPE

17:30-18:00 AWARDSANDCLOSINGCEREMONY


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Ø LECTURES


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S-1

REGULATIONOFRIPKSINCELLSURVIVALANDCELLDEATHBYAPOPTOSISANDNECROPTOSIS,INSIGHTSANDTHERAPEUTICPOTENTIAL

PeterVandenabeele1,2,3

1VIBInflammationResearchCenter,Technologiepark927,Zwijnaarde-Ghent,9052,Belgium

2DepartmentofBiomedicalMolecularBiology,GhentUniversity,Technologiepark927,Zwijnaarde-Ghent,9052,Belgium

3Methusalemprogram,GhentUniversity,Technologiepark927,Zwijnaarde-Ghent,9052,Belgium

Necroptosiswasinitiallyidentifiedasabackupcelldeathprogramwhenapoptosisisblocked.However,itisnow recognized as a cellular defense mechanism against infections and is presumed to be a detrimentalfactorinseveralpathologiesdrivenbycelldeath.Necroptosisisaprototypicformofregulatednecrosisthatdependsonactivationof thenecrosome,which is aprotein complex inwhich receptor interactingproteinkinase (RIPK) 3 is activated. TheRIPhomotypic interactionmotif (RHIM) is the coredomain that regulatesactivation of the necrosome. To date, three RHIM-containing proteins have been reported to activate thekinaseactivityofRIPK3withinthenecrosome:RIPK1,Toll/IL-1receptordomain-containingadaptorinducingIFN-β(TRIF),andDNA-dependentactivatorofinterferonregulatoryfactors(DAI).

RIPK1 is a keymolecule determining cellular fate downstream of several innate immune receptors. It is aserine/threonine kinase consisting of an N-terminal kinase domain linked by a largely unstructuredintermediate domain to a C-terminal death domain. In the TNF signaling pathway, RIPK1 paradoxicallypromotescellsurvivalaswellascelldeath.Theseoppositecellularfunctionsaremediatedby2distinctfacesof RIPK1. Upon binding of TNF to TNFR1, RIPK1 is recruited to the receptor complex I where it acts as ascaffold protein promoting cell survival, in part, by activating the canonical NF-kB pathway. SpecificconditionscanhoweveractivateRIPK1,anditskinaseactivitythenregulatesassemblyof2possiblecytosolicdeath-inducingcomplexes,namelycomplexIIb(RIPK1-FADD-Caspase-8)andthenecrosome(RIPK1-RIPK3-MLKL). These complexes respectively drive TNF-mediated apoptosis or necroptosis. The precisemolecularmechanism(s) controlling RIPK1 activation is (are) currently unknown. Similarly, how RIPK1 kinase activitycontributestobothcelldeathprocessesstillremainsunclear.Despitethislackofunderstanding,itisevidentthatRIPK1canplayadual roledownstreamofTNFR1and that itskinaseactivityneeds tight repression toavoidunnecessarydamagetotheorganism.

Targetingnecroptosiscanoccuratthreelevels:blockingRIPK1andRIPK3kinaseactivity,andblockingMLKL.Noveldrugsandknowndrugshavebeenidentifiedincellularscreeningwhichblocknecroptosis.Invivotheyareeffectiveinblockinginflammatory,degenerativeandinfectiousdiseases.Ontheotherhandinductionofnecroptosishasbeenfoundeffectiveininducingimmunogeniccelldeath.


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TYPE2TRANSGLUTAMINASE:AKEYREGULATOROFPROTEOSTASISUNDERCELLULARSTRESSFULCONDITIONS

MauroPiacentini

DepartmentofBiology,UniversityofRome“TorVergata”,Rome,Italy.

NationalInstituteforInfectiousDiseases,IRCCS“LazzaroSpallanzani”,Rome,Italy.Eukaryoticcellsareequippedwithanefficientqualitycontrolsystemtoselectivelyeliminatemisfoldedanddamaged proteins, and organelles. Abnormal polypeptides that escape from proteasome-dependentdegradation and aggregate in the cytosol can be transported via microtubules to inclusion bodies called'aggresomes',wheremisfoldedproteinsareconfinedanddegradedbyautophagy.WeshowedthatType2transglutaminase (TG2) knockout mice display impaired autophagy and accumulate ubiquitinated proteinaggregates upon starvation. Furthermore, p62-dependent peroxisome degradation is also impaired in theabsenceofTG2.Wealsodemonstratethat,undercellularstressfulconditions,TG2physicallyinteractswithp62and theyare localized in cytosolicproteinaggregates,whichare then recruited intoautophagosomes,whereTG2isdegraded.Interestingly,theenzyme'scrosslinkingactivityisactivatedduringautophagyanditsinhibition leadstotheaccumulationofubiquitinatedproteins.Takentogether, thesedata indicatethattheTG2 transamidating activity has an important role in the assemblyof protein aggregates, aswell as in theclearanceofdamagedmitochondriabymacroautophagy.Recently, we have isolated and characterized exosomes derived from cells either expressing or not TG2,under stressful conditions (i.e. proteasome impairmentor expressingamutated formofhuntingtin (mHtt)containing 84 polyglutamine repeats). Our results show that TG2 is present in the exosomes only uponproteasomeblockade,aconditioninwhichTG2interactswithTSG101andALIX,twokeyproteinsinvolvedinexosomebiogenesis. Interestingly,we found thatTG2 favours theassemblyofaproteincomplex includingmHtt, ALIX, TSG101 and BAG3, a co-chaperone involved in the clearance of mHtt. The formation of thiscomplex is paralleled by the selective recruitment of mHtt and BAG3 in the exosomes derived from TG2proficientcellsonly.Overall,ourdataindicatethatTG2isanimportantplayerinthebiogenesisofexosomescontrollingtheselectivityoftheircargounderstressfulcellularconditions.TakentogetherthesedataindicatethatTG2playsakeyroleintheregulationofproteostasisunderstressfulcellularconditions.


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S-3

AUTOPHAGYATTHEINTERSECTIONBETWEENCELLSURVIVALANDCELLDEATH:ROLESININFLAMMATIONANDLYSOSOMALHOMEOSTASIS

VojoDeretic

DepartmentsofMolecularGeneticsandMicrobiology,CellBiologyandPhysiologyandNeurology,University

ofNewMexicoHealthSciencesCenter,USA

Autophagy is a fundamental biological process that fulfills general and specialized roles in cytoplasmichomeostasis, and is at the intersection between cell survival and cell death. This talk will cover thesubsystems in autophagyand the recentprogress inourunderstandingof how they come together in thecontestof immunityandinflammation.Wewillalsogiveanupdateonorganizersofprecisionautophagyinthe context of immune and other functions. Furthermore, the role of TRIM proteins in autophagy-basedlysosomalhomeostasisandtheirroleininlysosomalcelldeathwillbepresented.


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NOVELREGULATORSOFAUTOPHAGY*

DevrimGözüaçık,MDPhD

SabancıUniversity,MolecularBiology,GeneticsandBioengineeringProgram,Tuzla,Istanbul,TURKEY.Correspondance:[email protected]

Web:http://myweb.sabanciuniv.edu/dgozuacik/Autophagyiskeybiologicaleventthatoccursatlowbasallevelsinallcelltypesfromyeasttomammalsundernon-deprivedconditions,performinghomeostatic functionssuchasproteindegradationandorganelle (e.g.mitochondria) turnover. It is rapidly upregulated during cellular stress, providing cells with recycledintracellular building blocks and substrates for energy generation, hence allowing them to surviveunfavorable conditions.Autophagy dysregulations play a critical role in the pathogenesis and progress ofseveral human health problems, including neurodegenerative disorders (i.e. Alzheimer's, Parkinson's andHuntington's diseases), degenerative syndromes (i.e. Dystrophies and dystrophic syndromes), lysosomalstoragedisorders(i.e.Gaucher'sdisease),inflammationandcancer.In Gozuacik Laboratory in Sabanci University, we mainly focus on the discovery of novel autophagyregulators:RNAs,proteinsandpathways(basicresearch).Moreoverinclosecollaborationwithclinicians,westudy implications of our findings in human disease formation (pathology and pathogenesis research) anddiagnosis (marker research). In collaboration with chemists and pharmacologists, we search formeans tomodulate autophagy for treatment purposes (drug research). In this speech, results from our basic andmedicalstudiesonautophagywillbediscussed.*Thisworkwas supported by The Scientific and Technological Research Council of Turkey (TUBITAK) 1001Grantnumbers112T272and110T405andSabanciUniversity.SelectedReferences:1) ArachicheA andGozuacikDx. Autophagy in health and disease. In the book: Toxicity and autophagy inneurodegenerativedisorders.JoseManuelFuentes(Ed.).SpringerPublishing.2015.ISBN:978-3-319-13938-8.2)TekirdagKA,OzturkDG,GozuacikDx.RegulationofautophagybymiRNAs.Inthebook:Autophagy:Cancer,Other Pathologies, Inflammation, Immunity, and Infection. HayatMA (Ed.). Elsevier Academic Press. 2015.ISBN:97801280103273)KorkmazG*,TekirdagKA*,OzturkDG,KosarA,SezermanOUandGozuacikDx.MIR376Aisaregulatorofstarvation-inducedautophagy.PLoSONE,2013,8(12):e82556.doi:10.1371/journal.pone.0082556.4) Tekirdag AK*, Korkmaz G*, Ozturk DG, Agami R, Gozuacik Dx. miR-181a regulates starvation- andrapamycin-inducedautophagythroughtargetingofATG5.Autophagy,2013March;9(3):1-12.5)OralO*, Oz-ArslanD*, ItahZ,NaghaviA,DeveciR,KaracaliS,GozuacikDx.CleavageofAtg3proteinbycaspase-8regulatesautophagyduringreceptor-activatedcelldeath.Apoptosis,2012Aug;17(8):810-20.6) Korkmaz G, le Sage C, Tekirdag AK, Agami R, Gozuacik Dx. miR-376b controls starvation and mTORinhibition-relatedautophagybytargetingATG4CandBECN1.Autophagy,2012February;8(2):165-176.


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S-5

MITOCHONDRIALSUBSTRATES:ATOOLTOCOMBATCANCER

BorisZhivotovsky

InstituteofEnvironmentalMedicine,DivisionofToxicology,KarolinskaInstitutet,Stockholm,Sweden;LomonosovMoscowStateUniversity,Moscow,Russia

Mitochondria play an important role in regulation of various cell death modalities. Outer mitochondrialmembranepermeabilizationandreleaseofseveralproteins,suchascytochromec,SMAC,AIF,etc.fromtheintermembranespaceofmitochondriaareregardedasa“pointofnoreturn”inmanymodelsofapoptosis.Accumulatingevidencesuggested thatmitochondria-generated reactiveoxygenspeciesare involved in thisprocess. However, depending on their overall concentration at steady state levels, and efficiency ofmitochondrialantioxidantenzymescelldeathcanbeprevented.Cancer cells demonstrate dramatically increased glycolysis even under air-saturated conditions (Warburgeffect),whereasmitochondrialcontributiontoATPsupplyisrestrained.Consequently,drugsthatcanperturbglycolysis,mightdisplaybeneficialtherapeuticeffects.Ourrecentobservationsrevealedthatamongagentsthat canmodulate tumor cell death aremembers of the Krebs cycle, succinate and citrate. The later candirectly suppress glycolysis via inhibition of phosphofructokinase. Importantly, mutations of succinatedehydrogenase (SDH)characterizeseveral tumors. InhibitionofSDHresults inaccumulationofsuccinate incytosol and subsequent activation of hypoxia-inducible factor, which is responsible for upregulation ofglycolyticpathwayandmitochondrial silencing.We found that inaddition to thispathway succinatemightsuppressapoptosisatmitochondriallevel.Alinkbetweenmitochondrialmetabolicchangesandcelldeathaswellashowalterationofenergyproducingpathwayscansensitizetumorcellstotreatmentwillbediscussed.


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S-6

AMBRA1NEGATIVECONTROLATTHECROSSROADAMONGAUTOPHAGY,CELLPROLIFERATIONANDCELLDEATH

FrancescoCecconi

DepartmentofBiology,UniversityofRomeTorVergata,Rome,Italy

CellStressandSurvivalUnit,DanishCancerSocietyResearchCentre,Copenhagen,DenmarkThe activating molecule in Beclin 1-regulated autophagy (AMBRA1), also known as autophagy/beclin-1regulator1, isahighly intrinsicallydisorderedandvertebrate-conservedadapterprotein that ispartof theautophagy signalling network. AMBRA1 is an important regulator of embryonic development, andboth itsmutationor inactivationhavebeenshown to impact severalpathologiesof thenervous system,and tobeinvolved in carcinogenesis. Recent studies have revealed that AMBRA1 can coordinate a cell response tostarvation or other stresses by integrated functions that include translocation of the autophagosome corecomplex to the ER, regulative ubiquitylation and stabilization of the kinase ULK1, selective mitochondriaremoval and cell cycle down-regulation. On the other side, AMBRA1 itself appears to be targeted by anumber of regulations, such as Cullin-dependent degradation, caspase cleavage and severalmodifications,rangingfromphosphorylationtoubiquitylation.HerewewilldiscusstworelevantnovelpathwaysofAMBRA1down-regulation:i) AMBRA1targetingbymiR7,anautophagy-andcell-cycle-relatedmicroRNA, inasignalling loopwehaveidentifiedthatinvolvestheoncogenec-mycandthephosphatasePP2A;ii)AMBRA1proteolytic cleavage togenerateanovelpositivemediatorofmitochondrial apoptosis. Indeed,theC-TerminalpartofAMBRA1,generatedbycaspasecleavageuponapoptosis induction, isableto inhibittheanti-apoptoticfactorBCL2byadirectbindingthroughitsBH3-likedomain.Altogether,bothmitochondrialAMBRA1-BCL2networkingandAMBRA1regulationbymiRNAsmayrepresentnoveltargetsindevelopmentoftherapeuticapproachesinhumandiseases.


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INVITROANDINVIVOCHARACTERIZATIONOFCELLSURVIVALGENESUSINGDESTABILIZEDCAS9

ŞerifŞentürk

DokuzEylulUniversity-IzmirInternationalBiomedicine&GenomeInstitute

One of the problems limiting the use of current CRISPR systems is the constitutive endonuclease activitywhen Cas9 and its sgRNA are co-expressed. Here, we sought to improve upon existing CRISPR/Cas9techniques to generate a system that (1) would provide potent, robust and temporally controlled geneediting, (2) be applicable to a broad spectrum of cell types and tissues, (3) facilitate high throughputmanipulationand (4)be traceable.To thisend,weexploited recentlydevelopedstrategies inwhichacell-permeable ligand is used in conjugation with a single genetically encoded destabilizing domain (DD) toregulatetheexpressionofanyproteinofinterest.ByfusingtheFKBP12-deriveddestabilizingdomaintoCas9(i.e., DD-Cas9) we demonstrated that this method of conditional regulation of protein stability could beexploitedforrapidandreversibleCas9expressioninvitro.Wevalidatedtheefficiencyofthisnewplatformbyconditionallytargetingavarietyofgenescontrollingdiversebiologicalprocesses.BytargetingtheRPA3geneandEGFR in the“EGFR-addicted”cells, inparticular,wedemonstrated theabilityof thesystemto identifygenes that are essential for sustained cell growth and survival. The unique aspect of this method is theconditionalregulationofCas9proteinexpressionindependentlyofitsmRNAexpression.WhencoupledwithaconditionalCreallele(Cre-ERT2),DD-Cas9couldbeutilizedtofacilitatetheanalysisofgenesthatmodulatediseaseonsetandprogressioninavarietyofpre-existingmousemodelsofhumandiseasebasedonCre-loxsystem. Insummary,ourdata indicatethatfusingCas9toadestabilizingdomainprovidesahighlyefficientandpotent,easyscalable,robustandtunablenewmodalityfortemporalcontrolofgeneeditingthatcanbeapplicabletoabroadspectrumofinvitroandinvivomodels.


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S-8

STAMPINGPROLIFERATIONANDCELLDEATHINPROSTATECANCER

FahriSaatçioğlu

UniversityofOslo,TheFacultyofMathematicsandNaturalSciences,DepartmentofBiosciences,Norway

The six transmembrane protein of prostate (STAMP) family, also known as six transmembrane epithelialantigen of prostate (STEAP), have been implicated in prostate cancer (PCa). STAMP1 and STAMP2 proteinexpressionisincreasedinhumanPCacomparedwithbenignprostateandtheyregulatecentralproliferativesignalingpathwaysinPCacells invitroandinvivo.Inaddition,STAMP1andSTAMP2expressioninhibitscelldeathresultinginrobusttumorgrowth.Consistentwiththesefindings,drug-inducedtherapeuticsilencingofSTAMPsbysystemicnanoliposomal-siRNAdeliveryprofoundlyinhibitstumorgrowthinpreclinicalmousePCamodels.ThesedatasuggestthatSTAMPshaveakeyroleindetermininglifeanddeathdecisionsinPCaandthusmayserveasnoveltherapeutictargets.


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S-9ETSTRANSCRIPTIONFACTORS-CRITICALREGULATORSOFBRAINTUMORINITIATING

CELLPROLIFERATIONVSNEURODEGENERATION

ErayŞahin1,2,3,MelisSavaşanSöğüt1,2,IşılAksanKurnaz1

1GebzeTechnicalUniversity,DepartmentofMolecularBiologyandGenetics,MolecularNeurobiologyLab(AxanLab),Gebze,Kocaeli

2YeditepeUniversity,BiotechnologyGraduateProgram,Kayisdagi,Istanbul3Presentaddress:AnadoluSaglikMerkezi,TıbbiHizmetlerDirektörlüğü,Gebze,Kocaeli

Object: Elk-1, a member of the ETS superfamily of transcription factors, has long been associated withregulationofimmediate-earlygenesinresponsetomitogen-dependentMAPKpathwayactivation.However,itspresence inpost-mitoticneuronshadbeenan intriguingfact. Workfromour labandothershavesinceshownthatElk-1isimportantforsurvivalofneurons,andthatknockdownofElk-1triggerscelldeath,yetthemechanismthroughwhichElk-1mediatesthiswasunclear.MaterialandMethod:Acombinationofmicroarray,promoteranalysesandluciferaseassays,aswellasreal-time PCR and brain tumor initiating cell assays have been used to underpin the role of Elk-1 inneurodegenerationvstumorigenesis.Results: Microarray analysis of Elk-1 overexpression in SH-SY5Y cell lines have shown that a number ofapoptosis and autophagy-related genes were regulated by Elk-1, in addition to components of hypoxiasignaling pathway. More interestingly, however, quite a number of organogenesis- and stem cell-relatedgeneswerefoundtoberegulatedinresponsetoElk-1.Tovalidatetheresults,wehavecarriedoutaseriesofreal-timePCRandluciferaseassaystoshowthatthesegenesindeedareregulatedbyElk-1invariousbraintumorcelllines,andthatCD133+cellsexpresshigherElk-1transcript.Conclusion: In linewithprevious findingsthatElk-1 isessential inhumanembryonicstemcells (hESCs)andthat ERK2MAPK cooccupies various promoters of cell-cycle and pluripotency-associated genes, our studyshows that Elk-1 also regulates pluripotency-related promoters in brain tumor initiating cells. On thecontrary,T417-phosphorylationofElk-1wasreportedtobeassociatedwithinclusionbodiesinanumberofneurodegenerativediseases.Webelievethatthesetworesultsarenotcontradictory–ourhypothesisisthatin theabsenceofpropersurvivalsignaling,or in thepresenceofpro-apoptoticones,Elk-1 instead initiatestheapoptoticpathway,henceactingasachoice-pointbetweenlifeordeath.Keywords:ETS,Elk-1,neurodegeneration,braintumorinitiatingcell,tumorigenesis


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DIMINISHEDCYCLINDEPENDENTKINASEACTIVITYANDMTORARECRITICALINTHECELLDEATHDECISIONTHROUGHAFFECTINGSTATSIGNALLINGDIFFERENTLYINPROSTATE

CANCERCELLS

ElifDamlaArısan,ÇağrıGümüşkaptan,ÖzgeBerrak,PınarObakan-Yerlikaya,AjdaÇoker-Gürkan,NarçinPalavanÜnsal

İstanbulKulturUniversity,DepartmentofMolecularBiologyandGenetics,AtakoyCampus34156Bakirkoy

Istanbul,d.arisan@iku.edu.trProstatecanceristhesecondmostfrequentlydiagnosedcanceraswellasthesixthleadingcauseofdeathinmales with cancer worldwide. Androgens play a critical role in prostate cancer development. Howeverprostate cancer cellsmay progress androgen-independently that causes highermortality rates. Therefore,new therapeutic targets and clarification of their signaling pathways is critical in treatment of aggressiveprostatecancercases.Oneofthepromisinganti-cancerstrategyistheinhibitionofproliferatingcancercellsviatargetingcyclinsandcyclin-dependentkinases(CDKs)complexes,whichcausessupressionofcellsurvivalsignallingroutes.NewgenerationCDKinhibitorsroscovitine(CYC202,seliciclib)orpurvalanolinhibitsspecificCDK targets and thus prevents cell proliferation and induces apoptosis. In this study, purvalanol androscovitine was used to expose the mechanism underlying mTOR-related apoptotic and/or autophagicresponseandtounderstandrolesofmTORdependingon itssignalcascades inthecelldeathprocessesviamTORsilencedandrogenreceptor(AR)negativePC3,DU145andARpositiveLNCaPprostatecancercelllines.In PC3 and LNCaP cells, CDK inhibitors were used alone and with mTOR siRNA combination to scan andanalyze differentiation upstream and downstream targets of mTOR using Pathscan ELISA Assay. CDKinhibitors purvalanol and roscovitine affects activation of mTOR and mTOR-related kinases in a CDK-independentmanner. However purvalanol exertsmore potent inhibitory function than roscovitine. In PC3andLNCaPcelllines,mTORdeficiencycausesblockageofapoptoticprocessesinducedbyCDKinhibitors.Ontheother hand, regulationof STAT1 and STAT3proteins bymTOR seems todetermine apoptotic effets ofthosedrugs.IncreasedSTAT3Ser727phosphorylationlevelsbyCDKinhibitorsleadstodecreaseSTAT3-FoXO1and CDK5 activity. Diminished CDK5 activity then causes AR-STAT3 dissociation. Therefore, especiallydifferentiationinSTAT3expressionandphosphorylationstatusplayavitalroleinthemannerofregulationofcell survivalandcelldeathpathways signalling.However,CDK inhibitorsand their combinationwithmTORsiRNA leads to diverse STAT3 expression profiles in DU145 cell line. In conclusion, CDK inhibitors arepromisingdrugcandidatesinthetreatmentofaggressiveprostatecancercellsthroughmodulatingdifferentmoleculartargetsdependsonthecelltype.


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S-11

THEBRIGHTANDTHEDARKSIDESOFREPROGRAMMINGTOPLURIPOTENCY

AndrasNagy

MountSinaiHospital,Lunenfeld-TanenbaumResearchInstitute,Toronto,CanadaSomaticcellreprogrammingwithafewdefinedtranscriptionfactorstopluripotencyisaseveralweekslongprocess.Thedrivingforcesbehindthisphenomenonandthecascadeofeventsareverypoorlyunderstood.Itishowever crucial touncover the finedetailsof thisprocess inorder to comprehend the truepropertyofthese inducedpluripotent stemcells (iPSCs)andsobetter tailor their future therapeuticanddiseasestudyuse.Several years ago, we developed a reprogrammingmethod utilizing apiggyBac (PB) transposon-mediateddeliveryof the reprogramming transgenes.Beyond theabilityof seamless removalof the transgenesoncepluripotentstemcellshavebeengenerated,thissystemhasadditionaluniquefeatures.Forexample,whencombinedwiththedoxycycline inducibletransgeneexpressionsystem,wefoundthatthesetransgenesareveryefficientlyregulatablebyaddingorwithdrawingdoxycycline.Invivodifferentiatedsomaticcellsderivedfrom these iPSCs canbe reprogrammed to “secondary” iPSCs (2ºiPSc) by simply addingdoxycycline to theculturemedium.Somaticcell linesproducedwiththismethodfrequentlyreturnto2ºiPScina"population"manner, which allows us to study the cascade of molecular events during the entire process ofreprogramming. This unexpected and unique property of the PB reprogramming system allowed thecharacterizationof the threeweek reprogrammingprocess at an almost daily resolution atmultiple omicslevels, leading toabetterunderstanding themolecularevents associatedwithgeneratingpluripotent cellsfromsomaticcells.In parallel, we have investigated the genetic changes associated with the reprogramming process. Weidentifieddenovogeneratedcopynumbervariationsattheearlyphaseofreprogramming,whichcreatedahigh level of genetic mosaicism. Intriguingly, the genome damage load is decreasing when the cells areculturedforanintermediateperiodoftime.Ourstudiesledustoconcludethatthisphenomenonisduetoselectionagainstmutatedcells.


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S-12

STEMCELLTHERAPYAPPROACHESTOISCHEMICCARDIOMYOPATHY

AlpCan

AnkaraUniversitySchoolofMedicineDepartmentofHistologyandEmbryologyLaboratoriesforStemCellsandReproductiveBiology

Sıhhiye,Ankara

Over the past 15 years, numerous stem cell trials have been performed in patients with ischemiccardiomyopathy (ICM), using both autologous and allogeneic stem cells. Althoughmany individual studiesreported encouraging signals, these were all phase 1 or 2 studies with appropriately small numbers ofpatients,andtheirconclusionsmustthereforebeconsideredpreliminary.Inanattempttoincreasestatisticalrobustness,arecentmeta-analysisassessingtheresultsofallrandomizedclinicaltrialsofstemcelltherapyforpatientswithacutemyocardial infarction(AMI)wasperformed,demonstratingnonetbeneficialeffectsonoutcomes,exceptforasmallimprovementinejectionfraction.Giventheseresults,areassessmentoftherationalefortheuseofstemcellsincardiovasculardiseaseistimely.PatientswithICMinvariablyhaveusuallyextensiveareasofmyocardialscar.ICMpatientshadareasofmyocardialdysfunctionduenottoscar,buttodysfunctional viablemyocardium (DVM). DVM provides a potential target for therapeutic interventions inICM. If the dysfunctional tissue consists of viable rather than scarred myocardium, LV function canpresumablybe improved.TheconceptofDVMmayalsohelpdirectwhichpatientsmaybenefitmost fromstemcell therapy.As thestageofHF thatmaybeconsidered too late for stemcell therapy isunclear, thepresenceofDVMmayhelpguidetheidentificationofthosepatientswiththemostpotentialtobenefit.Thepotentialofanytherapy,includingstemcells,toimproveoutcomesinICMisrelatednotonlytoitseffectsonrestoring function to DVM, but also to its capacity to improve processes that contribute to progressivedeteriorationofLVstructureandfunction.Onthebasisofthisconceptualframework,myaimistoexplorethepotentialof stemcells toexertbeneficialeffects in ICMbyconsidering theoverlapbetweenpathwaysbelieved to contribute to disease progression (other than atherosclerotic disease of the large coronaryarteries)andtheknownactivitiesof stemcells thatcould favorably influence thesepathways. (ProjectNo:0741-STZ-2014)


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TARGETINGINFLAMMATORYANDAPOPTOTICPATHWAYSBYAGENTSDESIGNEDBYMOTHERNATUREFORPREVENTIONANDTREATMENTOFCANCER

BharatB.Aggarwal,Ph.D.

FoundingDirector,Anti-inflammationResearchInstitute,SanDiego,California;USAFormerProfessorofExperimentalTherapeutics,CancerMedicineandImmunology,

TheUniversityofTexasM.D.AndersonCancerCenter,Houston,TexasPhone:832-754-0059;Email:[email protected]

Chronic infections, obesity, alcohol, tobacco, radiation, environmental pollutants, and high-caloriediethavebeenrecognizedasmajorriskfactorsforthemostcommontypesofcancer.Alltheseriskfactorsare linkedtocancerthrough inflammationandapoptosis.Whileacute inflammationthatpersists forshort-termmediateshostdefenseagainstinfections,chronicinflammationthatlastsforlong-termcanpredisposethehosttovariouschronicillnesses,includingcancer.Linkagebetweencancerandinflammationisindicatedby numerous lines of evidence; first, transcription factors NF-κB and STAT3, two major pathways forinflammation, areactivatedbymost cancer risk factors; second,an inflammatory conditionprecedesmostcancers; third, NF-κB and STAT3 are constitutively active in most cancers; fourth, hypoxia and acidicconditions found in solid tumors activate NF-κB; fifth, chemotherapeutic agents and gamma irradiationactivate NF-κB and lead to chemoresistance and radioresistance; sixth, most gene products linked toinflammation, survival, proliferation, invasion, angiogenesis, and metastasis are regulated by NF-κB andSTAT3;seventh,suppressionofNF-κBandSTAT3inhibitstheproliferationandinvasionoftumors;andeighth,most chemopreventive agents mediate their effects through inhibition of NF-κB and STAT3 activationpathways. Thus suppression of these proinflammatory pathways may provide opportunities for bothpreventionandtreatmentofcancer.Wewilldiscussthepotentialofnutraceuticalsderivedfromspicesandfromtraditionalmedicineinsuppressionofinflammatorypathwaysandtheirroleinpreventionandtherapyofcancer. SelectedReferences:

1. NF-kappaBandcancer:howintimateisthisrelationship.PrasadS,RavindranJ,AggarwalBB.MolCellBiochem.2009Oct8.

2. Curcumin Inhibits COPD-Like Airway Inflammation and Lung Cancer Progression in Mice.MoghaddamSJ,BartaP,MirabolfathinejadSG,Ammar-AouchicheZ,TorresGarzaN,VoTT,NewmanRA,AggarwalBB,EvansCM,TuvimMJ,LotanR,DickeyBF.Carcinogenesis.2009Sep30.

3. Curcuminmodulates the radiosensitivity of colorectal cancer cells by suppressing constitutive andinducibleNF-kappaBactivity. SandurSK,DeorukhkarA,PandeyMK,PabónAM,ShentuS,GuhaS,AggarwalBB,KrishnanS.IntJRadiatOncolBiolPhys.2009Oct1;75(2):534-42.

4. Design of curcumin-loaded PLGA nanoparticles formulation with enhanced cellular uptake, andincreased bioactivity in vitro and superior bioavailability in vivo. Anand P, Nair HB, Sung B,KunnumakkaraAB,YadavVR,TekmalRR,AggarwalBB.BiochemPharmacol.2009Sep6.

5. Signal transducer and activator of transcription-3, inflammation, and cancer: how intimate is therelationship?.AggarwalBB,KunnumakkaraAB,HarikumarKB,GuptaSR,TharakanST,KocaC,DeyS,SungB.AnnNYAcadSci.2009Aug;1171:59-76.

6. Curcuminpotentiatestheantitumoreffectsofgemcitabineinanorthotopicmodelofhumanbladdercancer through suppression of proliferative and angiogenic biomarkers. Tharakan ST, Inamoto T,SungB,AggarwalBB,KamatAM.BiochemPharmacol.2009

7. Inflammation, a silent killer in cancer is not so silent! Aggarwal BB. Curr Opin Pharmacol. 2009Aug;9(4):347-50.

8. Inflammationandcancer:howfriendlyistherelationshipforcancerpatients?AggarwalBB,GehlotP.CurrOpinPharmacol.2009Aug;9(4):351-69.

9. ZerumboneenhancesTRAIL-inducedapoptosis throughthe inductionofdeathreceptors inhumancolon cancer cells: Evidence foranessential roleof reactiveoxygen species. YodkeereeS, SungB,LimtrakulP,AggarwalBB.CancerRes.2009Aug15;69(16):6581-9.


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10. Curcumin sensitizes human colorectal cancer to capecitabine by modulation of cyclin D1, COX-2,MMP-9, VEGF and CXCR4 expression in an orthotopic mouse model. Kunnumakkara AB,DiagaradjaneP,AnandP,KuzhuvelilHB,DeorukhkarA,GelovaniJ,GuhaS,KrishnanS,AggarwalBB.IntJCancer.2009Nov1;125(9):2187-97.

11. Curcuminandcancercells:howmanywayscancurrykilltumorcellsselectively?RavindranJ,PrasadS,AggarwalBB.AAPSJ.2009Sep;11(3):495-510.

12. Acetyl-11-keto-beta-boswellic acid inhibits prostate tumor growth by suppressing vascularendothelialgrowth factor receptor2-mediatedangiogenesis.PangX,YiZ,ZhangX,SungB,QuW,LianX,AggarwalBB,LiuM.CancerRes.2009;69(14):5893-900.

13. Fabrication and characterization of silk fibroin-derived curcumin nanoparticles for cancer therapy.GuptaV,AsehA,RíosCN,AggarwalBB,MathurAB.IntJNanomedicine.2009;4:115-22.

14. K-Raspromotes angiogenesismediatedby immortalizedhumanpancreatic epithelial cells throughmitogen-activatedproteinkinasesignalingpathways.MatsuoY,CampbellPM,BrekkenRA,SungB,OuelletteMM,FlemingJB,AggarwalBB,DerCJ,GuhaS.MolCancerRes.2009Jun;7(6):799-808.

15. Molecular targets of nutraceuticals derived from dietary spices: potential role in suppression ofinflammationandtumorigenesis.AggarwalBB,VanKuikenME, IyerLH,HarikumarKB,SungB.ExpBiolMed(Maywood).2009Aug;234(8):825-49.

16. Nuclear factor-kappa B links carcinogenic and chemopreventive agents. Ralhan R, Pandey MK,AggarwalBB.FrontBiosci(ScholEd).2009Jun1;1:45-60.

17. Modelsforpreventionandtreatmentofcancer:problemsvspromises.AggarwalBB,DandaD,GuptaS,GehlotP.BiochemPharmacol.2009Nov1;78(9):1083-94.

18. Curcumin circumvents chemoresistance in vitro and potentiates the effect of thalidomide andbortezomibagainsthumanmultiplemyelomainnudemicemodel.SungB,KunnumakkaraAB,SethiG,AnandP,GuhaS,AggarwalBB.MolCancerTher.2009Apr;8(4):959-70.

19. Zerumbone abolishes RANKL-induced NF-kappaB activation, inhibits osteoclastogenesis, andsuppresses human breast cancer-induced bone loss in athymic nude mice. Sung B, Murakami A,OyajobiBO,AggarwalBB.CancerRes.2009Feb15;69(4):1477-84.

20. Theguggulforchronicdiseases:ancientmedicine,moderntargets.ShishodiaS,HarikumarKB,DassS,RamawatKG,AggarwalBB.AnticancerRes.2008Nov-Dec;28(6A):3647-64.

21. Withanolide sulfoxide from Aswagandha roots inhibits nuclear transcription factor-kappa-B,cyclooxygenaseandtumorcellproliferation.MulabagalV,SubbarajuGV,RaoCV,SivaramakrishnaC,DewittDL,HolmesD, SungB, Aggarwal BB, TsayHS,NairMG. Phytother Res. 2009 Jul;23(7):987-92.Targetinginflammatorypathwaysforpreventionandtherapyofcancer:short-termfriend,long-termfoe.AggarwalBB,VijayalekshmiRV,SungB.ClinCancerRes.2009Jan15;15(2):425-30.

22. Boswellicacidblockssignaltransducersandactivatorsoftranscription3signaling,proliferation,andsurvivalofmultiplemyelomavia theprotein tyrosinephosphatase SHP-1.KunnumakkaraAB,NairAS,SungB,PandeyMK,AggarwalBB.MolCancerRes.2009Jan;7(1):118-28.

23. Pharmacological basis for the role of curcumin in chronic diseases: an age-old spicewithmoderntargets.AggarwalBB,SungB.TrendsPharmacolSci.2009Feb;30(2):85-94.

24. Buteinsuppressesconstitutiveandinduciblesignaltransducerandactivatoroftranscription(STAT)3activation and STAT3-regulated gene products through the induction of a protein tyrosinephosphataseSHP-1.PandeyMK,SungB,AhnKS,AggarwalBB.MolPharmacol.2009Mar;75(3):525-33.

25. Resveratroladdiction:todieornottodie.ShakibaeiM,HarikumarKB,AggarwalBB.MolNutrFoodRes.2009Jan;53(1):115-28.

26. Zerumbonedown-regulateschemokinereceptorCXCR4expression leadingto inhibitionofCXCL12-induced invasion of breast and pancreatic tumor cells. Sung B, Jhurani S, Ahn KS,Mastuo Y, Yi T,GuhaS,LiuM,AggarwalBB.CancerRes.2008Nov1;68(21):8938-44.

27. AggarwalBB,KunnumakkaraAB,HarikumarKB, TharakanST, SungB,AnandP.Potential of Spice-DerivedPhytochemicalsforCancerPrevention.PlantaMed.2008Jul8.

28. AggarwalBB,HarikumarKB.Potentialtherapeuticeffectsofcurcumin,theanti-inflammatoryagent,against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplasticdiseases.IntJBiochemCellBiol.2008Jul9.[Epubaheadofprint]

29. SethiG,SungB,AggarwalBB.TNF:amasterswitch for inflammationtocancer.FrontBiosci.2008May1;13:5094-107.

30. AggarwalBB.Thepast,presentandfutureofmulti-targetedcancertreatment"Naturally":Foodforthought.CancerLett.2008May24.[Epubaheadofprint]Noabstractavailable.


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31. Kunnumakkara AB, Anand P, Aggarwal BB. Curcumin inhibits proliferation, invasion, angiogenesisandmetastasisofdifferentcancersthroughinteractionwithmultiplecellsignalingproteins.CancerLett.2008May12.[Epubaheadofprint]

32. Anand P, SundaramC, Jhurani S, KunnumakkaraAB, Aggarwal BB. Curcumin and cancer: An "old-

age"diseasewithan"age-old"solution.CancerLett.2008Aug18;267(1):133-64.33. HarikumarKB,AggarwalBB.Resveratrol:amultitargetedagentforage-associatedchronicdiseases.

CellCycle.2008Apr;7(8):1020-35.Epub2008Feb15.34. GoelA,JhuraniS,AggarwalBB.Multi-targetedtherapybycurcumin:howspicyisit?MolNutrFood

Res.2008Apr2.[Epubaheadofprint]35. GoelA,KunnumakkaraAB,AggarwalBB.Curcuminas"Curecumin":Fromkitchentoclinic.Biochem

Pharmacol.2007Aug19;[Epubaheadofprint]36. Aggarwal B.B, C. Sundaram,N.Malani andH. Ichikawa, “Curcumin: The Indian solid gold”, in The

MolecularTargetsandTherapeuticUsesofCurcumininHealthandDisease(edbyB.B.Aggarwal,Y-J.Surh,S.Shishodia),SpringerPublishingCompany,NewYork.,2007,p.1-76.

37. AggarwalBB,ShishodiaS,SandurSK,PandeyMK,SethiG.Inflammationandcancer:howhotisthelink?BiochemPharmacol.2006Nov30;72(11):1605-21.

38. AggarwalBB.Nuclearfactor-kappaB:atranscriptionfactorforallseasons.ExpertOpinTherTargets.2007Feb;11(2):109-10.

39. Jagetia GC, Aggarwal BB. "Spicing up" of the immune system by curcumin. J Clin Immunol. 2007Jan;27(1):19-35.

40. Aggarwal BB, Sethi G, Ahn KS, Sandur SK, Pandey MK, Kunnumakkara AB, Sung B, Ichikawa H.Targeting signal-transducer-and-activator-of-transcription-3 for prevention and therapy of cancer:moderntargetbutancientsolution.AnnNYAcadSci.2006Dec;1091:151-69.

41. GarodiaP,IchikawaH,MalaniN,SethiG,AggarwalBB.Fromancientmedicinetomodernmedicine:ayurvedic concepts of health and their role in inflammation and cancer. J Soc IntegrOncol. 2007Winter;5(1):25-37.

42. AhnKS,AggarwalBB.TranscriptionfactorNF-kappaB:asensorforsmokeandstresssignals.AnnNYAcadSci.2005Nov;1056:218-33.

43. AggarwalBB,ShishodiaS.Moleculartargetsofdietaryagentsforpreventionandtherapyofcancer.BiochemPharmacol.2006;71(10):1397-421.

44. AggarwalBB,KumarA,BhartiAC.Anticancerpotentialofcurcumin:preclinicalandclinicalstudies.AnticancerRes.2003Jan-Feb;23(1A):363-98.

45. AggarwalBB.SignallingpathwaysoftheTNFsuperfamily:adouble-edgedsword.NatRevImmunol.2003Sep;3(9):745-56.

46. AggarwalBB.Nuclearfactor-kappaB:theenemywithin.CancerCell.2004Sep;6(3):203-8.47. Dorai T, Aggarwal BB. Role of chemopreventive agents in cancer therapy. Cancer Lett. 2004 Nov

25;215(2):129-40.48. AggarwalBB, TakadaY,OommenOV. Fromchemoprevention to chemotherapy: common targets

andcommongoals.ExpertOpinInvestigDrugs.2004Oct;13(10):1327-38.49. Aggarwal BB, Shishodia S. Suppression of the nuclear factor-kappaB activation pathway by spice-

derivedphytochemicals:reasoningforseasoning.AnnNYAcadSci.2004;1030:434-41.50. Aggarwal BB, Bhardwaj A, Aggarwal RS, SeeramNP, Shishodia S, Takada Y. Role of resveratrol in

prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res. 2004 Sep-Oct;24(5A):2783-840.

51. AhnKS,AggarwalBB.TranscriptionFactorNF-{kappa}B:ASensorforSmokeandStressSignals.AnnNYAcadSci.2005Nov;1056:218-33.

52. Shishodia S, Sethi G, Aggarwal BB Curcumin: getting back to the roots. Ann N Y Acad Sci. 2005Nov;1056:206-17.

53. GargAK,Buchholz TA,AggarwalBB.Chemosensitizationand radiosensitizationof tumorsbyplantpolyphenols.AntioxidRedoxSignal.2005Nov-Dec;7(11-12):1630-47.Review.

54. AggarwalBB,ShishodiaS,TakadaY,Jackson-BernitsasD,AhnKS,SethiG,IchikawaH.TNFblockade:aninflammatoryissue.ErnstScheringResFoundWorkshop.2006;(56):161-86.

55. AggarwalBB,IchikawaH,GarodiaP,WeerasingheP,SethiG,BhattID,PandeyMK,ShishodiaS,NairMG.FromtraditionalAyurvedicmedicinetomodernmedicine: identificationoftherapeutictargetsforsuppressionofinflammationandcancer.ExpertOpinTherTargets.2006Feb;10(1):87-118.


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56. AggarwalBB,ShishodiaS,SandurSK,PandeyMK,SethiG.Inflammationandcancer:Howhotisthelink/BiochemPharmacol.2006Nov30;72(11):1605-21

57. Aggarwal, B.B. and Shishodia S., Surh Y-J (eds.) The Molecular Targets and Therapeutic Uses ofCurcumininHealthandDiseaseSpringerPublishers,NewYork,2007

58. Aggarwal,B.B.andShishodiaS.,(eds.)Resveratrol inHealthandDisease.TaylorandFrancisBooks,Inc.,Boston,pp.1-679,2006


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MOLECULARMECHANISMSOFFLAVONOIDQUERCETININENHANCINGAPOPTOSISINCHRONICLYMPHOCYTICLEUKEMIA

GianLuigiRusso,MariaRusso,CarmelaSpagnuolo,IdoloTedesco,StefaniaMoccia

InstituteofFoodSciences,NationalResearchCouncil,83100Avellino,Italy;e-mail:[email protected]

Quercetinisthemostabundantflavonoidpresentinthedietanditsdiseasepreventingpropertieshavebeenlargely investigated. Among these, quercetin is able to modulate several hallmarks of cancer, includingresistancetoapoptosis.Previousstudiesfromourgroupdemonstratedthecapacityofquercetintosensitizeseveralleukemiacell linesandB-cells isolatedfrompatientsaffectedbychroniclymphocyticleukemia(CLL)todeathligandagonists(e.g.,anti-CD95andrTRAIL).Moreover,inassociationwithcanonicalandinnovativechemotherapeutic drugs (fludarabine, ABT-737 and BH3-mimetics), quercetin synergistically enhances thedrugresponseagainstCLL.Thiseffectismediatedbychangesintheexpressionandactivityofanti-apoptoticproteins belonging to theBcl-2 family. Among these,Mcl-1 has been associated to apoptotic resistance inCLL.Aimofthispresentationistoreviewtheapoptotic-enhancingactivityofquercetininvitro(leukemiccelllines)andexvivo(B-CLLcells)models,dependinguponthedown-regulationofMcl-1.Usingarelativelynewcell line, HG3 derived from primary B-cells immortalized with Epstein-Barr virus, we will show how theassociationbetweenBH3-mimeticsandquercetinsynergistically inducesapoptosisthroughtheinhibitionofPI3K/Akt signaling pathway.We also identified the protein kinase CK2 as the direct and primary target ofquercetin,sinceCK2activityisinhibitedbytheflavonoidwithinoneminutefromthetreatment.InthecaseofABT-737,aBH3-mimeticcompoundwhichbindsandinactivatesBcl-2,Bcl-XL,BCL-Wmembers,butweaklyinteractswithMcl-1,wedemonstratedthatthecombinedtreatmentofABT-737andquercetinrepresentsapromising new therapeutic strategy against B-CLL. In fact, since quercetin contributes to remove theresistance due toMcl-1 over-expression, it allows ABT-737 to exhibit its therapeutic efficacy against pro-survivalBcl-2factors.Consideringtherapiduptakeofquercetinanditslowtoxicityagainstnormalperipheralbloodcells,wearedesigningaclinicalstudyonCLLpatientsaimedtodemonstratethepotentialuseofthemoleculeintheadjuvantchemotherapyagainstCLL.


4-7

May2

016

S-15

DevelopmentOfNovelTargetedTherapiesForSolidTumors

BülentÖzpolat

DepartmentofExperimentalTherapeuticsandCenterforRNAInterferenceandNon-CodingRNA,TheUniversityofTexasM.D.AndersonCancerCenter,Houston,Texas


4-7

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016

S-16

MERRECEPTORTYROSINEKINASEANDMACROPHAGEPHAGOCYTOSISOFAPOPTOTICCELLS

IanDransfield,NicoleD.Barth,JohnA.Marwick,GregLemke,MaryJoHeeb,ErinD.Lew,&Adriano

G.RossiOne of the hallmark changes associated with apoptosis is the exposure of anionic phospholipids such asphosphatidylserine (PtdSer) and phosphatidylethanolamine on the outer leaflet of the plasmamembrane.Phagocytic cells express a variety of receptors that confer the capacity for recognition of this membranealteration, including BAI-1 and Tim4. In addition, phagocytes express receptors that interact with PtdSer-bindingmoleculesthatactasabridgebetweenthephagocyteandtheapoptoticcell.Mer(genenameMertk)is a receptor tyrosine kinase expressed by a variety of different cell types, includingmacrophages derivedfrommost tissues.Mer serves as a receptor for Protein S and Gas6, PtdSer binding proteins that rapidlyopsoniseapoptoticcells.MercanmediatetetheringofProteinS-opsonisedapoptoticcellsandactivationofthe intrinsic kinase activity of Mer signals apoptotic cell internalisation. In addition, TLR-dependent pro-inflammatory cytokine production by macrophages is influenced by Mer-dependent signalling. We haveexaminedMerexpressionduringhumanmonocytedifferentiation tomacrophages invitroandexplore therelationship between monocyte expression of Mer and the capacity to bind the Mer ligand, Protein S.Together,ournewdatarevealanovelroleforMerinthecontrolofmonocytefunction.


4-7

May2

016

S-17UnequalCellDeathinDifferentiatingThelperCells

AytenNalbant

IzmirInstituteofTechnology,DepartmentofMolecularBiologyandGenetics,MolecularImmunologyand

GeneRegulationLaboratory,Urla,İzmir35430,Turkey*[email protected]

T lymphocytes play a pivotal role in the immune response as a key regulatory and effector cells. Uponantigenicstimulation,naiveCD4Tcellscanactive,proliferateanddifferentiateintouniquesignaturecytokineexpressing effector T helper (Th) cells for instance Th1, Th2 and Th17. Recently discovered Th17 cells areknown to play a critical role in various inflammatory pathologies including Multiple sclerosis (MS),Rheumatoid arthritis (RA) and cancer. Th1 cells are involved in cell-mediated immune response and areresponsiblefortheclearanceofintracellularpathogens.Th2cellsareinvolvedinhumoralimmuneresponses,allergies and responds to some parasites. They secrete IL-4, IL-13, IL-10 and IL-5. Th17 cells are linked toseveralautoimmunediseasesandmediateimmuneresponsetobacterialandfungalinfections.TheysecreteIL-17,IL-21andIL-22cytokines.Apoptosis is a process that involves sequential activation of a series of proteins such as caspases after anappropriate stimulation. Apoptosis can be carried out by external (receptor-ligand interaction) or internal(mitochondria)mediatedpathways.Fas,FasL,DR5andTRAILareapoptoticproteinswhereasFLIPandBcl-xLareanti-apoptoticproteins,whichinvolveinapoptoticpathways.ItisknownintheliteraturethatActivationInduced Cell Death (AICD) is important in eliminating activated T cells. The Fas/FasL pathway is veryimportant in T cell death. TCR signaling can lead to the deletion of activated peripheral T cells throughapoptosis.ThesurvivalordeathofeffectorTcellsisaffectedonIFNγ,IL-4andIL-2.Thelpercellsshowdifferentsusceptibility toapoptosis.For instance,Th17cellsare lesssensitivethanTh1cellstoFasmediatedapoptosisbecausetheyhaveahigherexpressionofFLIP.Th17aremoresensitivetoFasmediatedapoptosisthanTh2cellsduetotheirhigherexpressionofFasL.Re-stimulatedTh17cellsundergoAICDthroughaFas/FasLmediatedpathwaythatisunaffectedbyIFNγ.HumanTh17cellsarephenotypicallyresemblesdifferentiatedmemoryTcellsbuttheyaredistinctfromcentralmemory,exhaustedandsenescentT cells. Human Th17 cells are long lived cells. Overall data showed that there is an unequal cell death indifferentiating T helper subsets. The further investigation of cell survival and death signal networks in Thelper cells will help us better understand the molecular mechanisms of various pathologies such asautoimmunity, cancer and infection diseases. Thisworkwas supported by a grant from the Scientific andTechnological ResearchCouncil of Turkey (TUBITAK). Project grantnumber is 110T412andawarded toDr.AytenNalbant.Keywords:Tcells,Thelpers,Th1,Th2,Th17,AICDandapoptosis


4-7

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016

S-18

GabrielLOPEZ-BERESTEIN


4-7

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016

S-19

APOPTOSISASAPOWERFULMEANSOFIMMUNEMODULATIONFORTHETREATMENTOFTYPE1DIABETES

HavalShirwan

SchoolofMedicine,UniversityofLouisville,Louisville,Kentucky,USA

[email protected] 1 diabetes (T1D) is a debilitating autoimmune disease affecting a significant portion of populationworldwide. Insulin treatmentasstandardofcare isoften ineffective inpreventingrecurrenthyperglycemicepisodeswith long-termundesiredadverseeffects.Transplantationofpancreatic isletsasa sourceofbetacells producing insulin has proven effective in improving metabolic control/quality of life and preventingsevere hypoglycemia in patients with T1D. However, islet transplantation suffer from immune rejectionirrespectiveofchronicuseof immunosuppressionanditscomplications. BothautoimmunityandisletgraftrejectionareinitiatedandperpetuatedbyanimbalancebetweenpathogenicTeffector(Teff)andprotectiveTregulatory(Treg)cells.TeffcellsupregulateFasreceptorontheirsurfacefollowingactivationandbecomesensitive to Fas/FasL-mediated apoptosis. Therefore, Fas-mediated apoptosis has a great potential tospecificallypurgeoutautoandallo-reactivepathogenicTeffcells for thepreventionandtreatmentofT1D.WehavegeneratedanovelformofFasLproteinwithimprovedapoptoticactivityanddemonstrateditsutilityfor the induction of tolerance to auto and allogeneic antigens for the treatment of T1D in various rodentmodels. Tolerance was initiated by a regulatory circuit involving apoptosis of Teff cells, phagocytosis,secretionofTGFβ,andgeneration/expansionofTregulatorycells.Tregulatorycellswerenotonlyimportantin tolerance induction but also maintenance. This concept is presently being pursued for clinicaldevelopment.


4-7

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016

S-20

ABNORMALBRAINGANGLIOSIDEACCUMULATIONTRIGGERSAPOPTOSISINEARLYONSETTAY-SACHSDISEASEMOUSEMODEL

VolkanSeyrantepe

IzmirInstituteofTechnology,DepartmentofMolecularBiologyandGenetics,Izmir,Turkey

volkanseyrantepe@iyte.edu.trTay-SachsdiseaseisaseverelysosomalstoragedisordercausedbymutationsintheHEXAgenecodingforαsubunitoflysosomalβ-HexosaminidaseAenzyme,whichconvertsGM2toGM3ganglioside.Unexpectedly,theHexA-/-micehaveanormallifespanandshownoobviousneurologicalimpairmentsuntilatleast1yearofage,owingto theabilityof thesemicetocatabolizestoredGM2gangliosideviasialidase(s) removingsialicacidintoglycolipidGA2whichfurtherprocessedbyβ-HexosaminidaseB,therebybypassingtheHexAdefect.To elucidate whether sialidase Neu3 can contribute to GM2 ganglioside degradation, we generated micemodelwithcombineddeficienciesofβ-HexosaminidaseAandSialidaseNeu3.HexA-/-Neu3-/-micearehealthyat birth but died at 1.5-4.5months of age. Thin layer chromatography andmass spectrometry analysis ofbrain from HexA-/-Neu3-/- mice showed abnormally accumulated GM2 ganglioside level. Slow movement,ataxiaandtremorareamongneurologicalabnormalities.Inthecurrentstudy,wedelineatewhetherthereisapoptosis in Tay-Sachs mice model’s brain. In order to profile the expression of 84 key genes related toapoptosis in the cerebellum and cortex from 4.5months oldmice,we used RT2ProfilerPCRArray systemspecific for apoptosis. We found that mRNA levels of pro-apoptotic genes such as TNF and caspase 4increased 5.8X and 2.1X respectively in HexA-/-Neu3-/-. On the other hand, mRNA levels of anti-apoptoticgenessuchasBag1andBcl2(1.6X),Cd40lg(1.8X)andNaip(7.1X)decreasedincerebellumofthesamemiceascomparedtoHexA-/-micerevealingtheapoptosisinearly-onsetTay-Sachsmicemodel. Wesuggestthatonce a critical threshold of GM2 ganglioside storage is reached in the cerebellum, a signaling cascade istriggeredwhichactivatesneuronaldeath.


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016

Ø ORALPRESENTATIONS


4-7

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016

OP-1

ANDROGENRESPONSEISREQUIREDFORDNADAMAGERESPONSEINOXIDATIVESTRESSCONDITIONS

BilgeDebeleçBütüner¹,KemalSamiKorkmaz²

¹|EgeUniversity,FacultyofPharmacy,Dept.ofPharmaceuticalBiotechnology,35100Bornova,Izmir,Turkey²|EgeUniversity,FacultyofEngineering,Dept.ofBioengineering,35100Bornova,Izmir,Turkey

Aim: As it has been shown that inflammatorymicroenvironment leads to the proteosomal degradation ofandrogenreceptor (AR),prostatic inflammationwasassociatedwithdevelopmentofcarcinoma inpreviousstudies.Inaddition,androgensignalingalsocontributestodistinctcellularmechanismsincludingantioxidantresponse regulation. To investigate the role of androgens in oxidative stress conditions in prostate cellsoxidativeDNAdamageandsubsequentdamageresponsewasexamined.MaterialandMethod:We used amodel of inflammatorymicroenvironment tomimic the chronic inflammation and/oroxidativestressconditionsbytreatingprostatecellswithinflammatoryconditionedmedia(CM)withknowncytokine concentrations or H2O2. Then, the cellular responses on the ROS generation and DNA damageresponsewasinvestigated.

Results:Wedemonstratedthatthetreatmentoftheprostatecellswithoxidativestressinducers(menadioneandH2O2) resultedwith increased formationof reactiveoxygen species (ROS) andoxidativeDNAdamageuponARdepletion.Consistently,whileDNAdamage (H2AXandAP sites)was increasedafterAR silencing,DNAdamagerecognitionwasdisrupted thatwasshownbydecreasedATMphosphorylations in thesecellssuggestingthatARsignalingisrequiredforDNAdamageresponseinprostatecancercelllines.Furthermore,androgen-independentLNCaP104r2cellswerefoundtobemoretoleranttooxidativestresswithhigherROSlevelsandactivatedSIRT1expressionresultingreducedoxidativeDNAdamage.

Conclusions: Thus, loss of AR function results in increased DNA damage upon abrogated DNA damageresponsemechanism. In addition, androgens has a significant role in controlling oxidative stress state andfurther damage response cascade of the cell. Therefore, though androgen ablation is known as themaintherapeuticapproachforprostatecancercure,lossofandrogensignalingseemstoinducefurtheroxidativestressresistanceandDNAdamagegenerationbypromotingtumorigenicalterationsinprostatecells.

Keywords:Androgenreceptor,oxidativestress,DNAdamage,γH2AX,prostatecancer


4-7

May2

016

OP-2

NEUROPROTECTIVEROLEOFCURCUMINONCNSDAMAGEINOFFSPRINGOFPREGNANTRATSEXPOSEDTOAROCLOR1254

MehmetErayAlcıgır¹,HalefOkanDoğan²,BegümYurdakökDikmen³,KübraDoğan⁴,SevilAtalayVural¹,FatmaMeriçYılmaz5,Atillaİsgören6

¹|AnkaraUniversity,FacultyofVeterinaryMedicine,DepartmentofPathology,Ankara/TURKEY²|CumhuriyetUniversity,FacultyofMedicine,DepartmentofBiochemistry,Sivas/TURKEY

³|AnkaraUniversity,FacultyofVeterinaryMedicine,DepartmentofPharmacologyandToxicology,Ankara/TURKEY

⁴|SivasNumuneHospital,DepartmentofBiochemistry,Sivas/TURKEY5|YildirimBayezitUniversity,FacultyofMedicine,DepartmentofBiochemistry,Ankara/TURKEY

6|AnkaraUniversity,FacultyofMedicine,LabaratoryAnimalUnit,Ankara/TURKEY

Inthisstudy,itisaimedtoidentifythedamagestothecentralnervoussysteminoffspringofratsexposedtoAroclor 1254, toxic Polychlorinated biphenyls (PCBs), during critical peri-od of pregnancy and, invivo andinvitro,torevealoutneuroprotectiveeffectofcurcumin,apowerfulantioxidant,onthethedamages.Inthestudy,3groupswereconstitutedand2fe-malesand1maleWistarratswerematedineachgroups.Newbornpups(n=10)wereran-domlyselectedfromeachgrups.Thefirst(control)groupwasallocatedwithoutanypro-cessingtopups.ThepregnantratsofsecondgroupwereexposedtoAroclor1254with1mg/kg/BWdoseduring7thto21stdaysofgestation.ThepregnantratsofthirdgroupwereexposedtodoseofAroclor1254withinsamedaysanddose.Theratpupsinlastgroupwerealsoorallytreatedwithcurcumindissolvedin50%DMSO with 200 mg/kg/BW dose during the first postnatal three days. The damages were comparedhistopathologicallyinthelasttwogroups.8-OHdG,4HNEtheMBPexpressionsandDNAinsitufragmentationpositivities (by TUNELmethod) were immunohistochemically evaluated and decided how curcumin to de-creasedatthose8OHdGand4HNEactivitiesandincreasedatMBPactivity.RatF98gliomalinewereappliedwithAroclor1254andcurcuminseparatelyandtogether.MTT,LDHandcytotoxicitytestwereperformedandevaluated statistically. Especially 8OHdG and TUNEL reactions gave similar results in prosenchephalon andmesencephalon. InvitrostudiesshowthatcurcuminreduceoncytotoxicityandDNAdamageincellsduetooxidativestressbyAroclor1254ongliomacells.Inconclusion,curcuminhasroleinreducingoxidativestressandreturningeffectsofDNAdamageprocessedbyAroclor1254. It isthoughtonneuroprotectiveeffectofcurcuminthatitshigherdosesmaybemoreeffectiveovertheentireCNS.

Keywords:offspringrat,Aroclor1254,curcumin,CNS,invivoandinvitro


4-7

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016

OP-3

PROTECTIVEEFFECTSOFDIFFERENTANTIOXIDANTSAGAINSTTHETOXICITYOF3,5-DIMETHYLAMINOPHENOLINHUMANBLADDERCELLLINE

PınarErkekoğlu¹'²,Ming-WeiChao¹'³,Chia-YiTseng4,BelmaKoçer-Gümüşel²,BevinP.Engelward¹,GeraldNWogan¹,StevenR.Tannenbaum¹

¹|DepartmentofBiologicalEngineering,MassachusettsInstituteofTechnology,Cambridge,MA02139,USA.

²|HacettepeUniversity,FacultyofPharmacy,DepartmentofToxicology,06100Ankara,Turkey³|DepartmentofBioScienceTechnology,CollegeofScience,ChungYuanChristianUniversity,Zhonglidistrict,

Taoyuancity,Taiwan3204|DepartmentofBiomedicalEngineering,CollegeofEngineering,ChungYuanChristianUniversity,Zhongli

district,Taoyuancity,Taiwan320

Monocyclic aromatic amines (MAAs) are widespread environmental contaminants with multiple sources.ExposuretoMAAsmainlycausetoxiceffectsonbladderandincreaseinbladdercancerincidence.Theirmainoxidativemetabolitesare theiro-orp-phenolderivatives.Asa leadingexample, thephenolicderivativeof3,5-dimethylaniline,namely3,5-dimethylaminophenol (3,5-DMAP), is readilyoxidized to thequinone iminefollowingadministrationandmammaliancellstreatedwiththiscompoundexhibitelevatedlevelsofreactiveoxygen species (ROS) for several days following exposure. The main aim of this study was to investigatewhether this compound caused an oxidant/antioxidant imbalance in human bladder cells (UROtsa cells).Furhermore, we investigated whether this compound causes DNA damage and increases in the caspases(caspase3and8),whicharemainenzymesthatplayessentialroleinapoptosis.Protectiveeffectsoforganic(selenomethionine) and inorganic (sodium selenite) selenocompounds, ascorbic acid and N-acetylcysteine(NAC)againstitstoxicitywasalsoassessed.Weobservedthatatinhibitoryconcentration(IC50)of3,5-DMAPwasapproximately50µM.Besides,3,5-DMAPcausedadosedependentincreaseofROS.Inbothcytoplasmandnucleus,3,5-DMAPcausedalterations intheantioxidantenzymeactivitiesanddecreasescellularredoxratio. Lipid peroxidation and as a consequence protein oxidation were elevated in particularly cytoplasm.Single strandDNA breakswere also induced by 3,5-DMAP; however no changeswere observed in doublestrandDNAbreaks.Moreover,apoptosiswasalsotriggeredby3,5-DMAPexposureasevidencedbyincreasesin caspase 3 and caspase 8 activities. Selenocompounds, ascorbic acid and NAC were found to be bothpartiallyprotectiveagainstthecellulartoxicityof3,5-DMAP.

Keywords:3,5-dimethylaminophenol,alkylaniline,reactiveoxygenspecies,cyctotoxicity,apoptosis


4-7

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016

OP-4

REDUCTIONOFLIVERCIRRHOSISBYTGF-ΒETATYPE1RECEPTORKINASEINHIBITOR,LY-364947

ÖzgeÇağlar,NecmiyeCanacankatan,FigenDoran

Object: Cirrhosis is a common and consequence of chronic liver disease that characterized by the diffusefibrosis, scar tissue and regenerative nodules. Transforming growth factor (TGF) beta type 1 is one of themajorprofibrogenicmediatorswhichtakespart inthedevelopmentof livercirrhosis.Thebalancebetweencellproliferationandapoptosisisveryimportantintheformationoflivercirrhosis.Inthisstudy,weaimedtodetermine the apoptotic and antioxidantmechanisms in liver cirrhosis and the effect of the TGF-β type Ireceptorkinaseinhibitor,LY-364947onlivercirrhosis.Material and Method: 6 groups were included as Control, Cirrhosis,Cirrhosis + DMSO, Cirrhosis + LY-364947(100μg/kg/week),Cirrhosis + LY- 364947(300μg/kg/week), after Cirrhosis + LY-364947(100μg/kg/week). Experimental liver cirrhosis was developed by N-Nitrosodiethylamine. Control,Cirrhosis, Cirrhosis + DMSO, Cirrhosis + LY- 364947(100μg/kg/week), and Cirrhosis + LY- 364947(300μg/kg/week) groups were killed when cirrhosis was established by evaluation of histopathologicalinvestigations inratswhichwerechosenrandomlyfromtheCirrhosisgroup.AftercirrhosiswasestablishedLY- 364947was administered for 4weeks to ratsCirrhosis+ LY- 364947 (100μg/kg/week) group.Apoptosiswas evaluated bymeasurement of caspase -3,-8 and -9. Antioxidant capacity and lipid peroxidationwereevaluatedbymeasuring the levelsof reducedglutathione (GSH)andmalondialdehyde (MDA), respectively.The evaluations were carried out by colorimetric methods according to the assay instructions.Histopathologicalinvestigationswerealsocarriedout.Results: Caspase -3,-8 and -9 enzyme activities and also increased lipid peroxidation were significantlysuppressedbyLY-364947(300μg/kg/week).Histopathological findings indicated that cirrhosisdevelopmentwas reduced by LY- 364947(300μg/kg/week) whereas administration of LY- 364947(100μg/kg/week) aftercirrhosiswas failed topointout thesameaffect.GSHdepletionwasobserved inallgroupscomparedwithcontrol.Conclusion:LY-364947maybeconsideredasapromisingantifibroticagentinlivercirrhosisbysuppressingapoptosisandlipidperoxidationandreducinglivercirrhosis.Keywords:livercirrhosis,LY-364947,TGF-βtypeI,caspase,apoptosis


4-7

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016

OP-5

THEABILITYOFTHYMOQUINONETOINDUCECASPASE-INDEPENDENTCELLDEATHINDIFFUSELARGEBCELLLYMPHOMAISMEDIATEDBYINTRACELLULARCALCIUMRELEASE

MimouneBerehab¹,RedouaneRouas¹,HaidarAkl²,DouaeMoussaAgha¹,ArsèneBurny¹,FabriceJourne³,GhanemGhanem³,DominiqueBron¹,PhilippeLewalle¹,MakramMerimi¹

¹|LaboratoryofExperimentalHematology,InstitutJulesBordet,CentredesTumeursdel’ULB,UniversiteLibredeBruxelles

²|LaboratoryofMolecularandCellularSignaling,DepartmentofCellularandMolecularMedicine,KULeuven³|LaboratoryofOncologyandExperimentalSurgery,InstitutJulesBordet,UniversiteLibredeBruxelles

Introduction: Disruption of the apoptotic pathways in diffuse large B cell lymphoma (DLBCL) remains achallenge for standard treatmentand results in refractorinessandpoorprognosis. Thymoquinone (TQ), anactive compound from amedicinal plantNigella sativa, has been reported to kills cancerous cells throughdifferentcelldeathpathways.Nevertheless,themolecularmechanismsunderpinningtheinterplaybetweendifferent cell deathmodalities are still unclear. Herein, we explored the in vitro anticancer activity of TQagainstDLBCLandtrendtounderstandthemechanismsofitsanticanceractivity.Results: TQ greatly inhibited the proliferation and induced cell death of DLBCL cell lines and primaryrefractoryDLBCLcellswithaminimaleffectonnormal lymphocytes.Molecular investigationsrevealedthatTQactivates theUPRpathway in highly responsive cell lines, identified through sXBP-1,GRP78, CHOPandHSPA1Aup-regulation.Inparallel,TQactivatedthemitochondrialpathwayofapoptosisrevealedbycaspase-3and -9activationconsecutive tocytochromec release.However themitochondrialeventsdon’tplayedacriticalroleinhighlyresponsivecells,infact,caspaseinhibitionbyz-VAD-fmkfailedtorescuethemfromTQ-induced cell death, while this was partially abrogated in the lowly responsive one. In contrast, cytosoliccalciumchelationbyBAPTA-AMstronglyrepressedTQcelldeatheffectinhighlyresponsivecelllines.Indeed[Ca2+]c measurement showed that TQ leads to an acute [Ca2+]c rise in highly responsive cells with amoderateriseinthelessresponsiveone.Theobserved[Ca2+]crisewasderivedmainlyfromtheERcalciumstores andmediatedby IP3 receptors, in fact their inhibitionby 2-APB strongly prevented TQ-induced celldeath.Conclusion:Thisworkdemonstratesrequirement[Ca2+]criseintheabilityofTQtoinducenon-apoptoticcelldeath, which may offer an effective strategy for antilymphoma therapy in DLBCL intrinsically resistant orrefractorytochemotherapyinducedapoptoticcelldeath.Keywords:TQ:ThymoquinoneER:endoplasmicreticulumUPR:unfoldedproteinresponse[Ca2+]c :cytosoliccalciumsXBP1:splicedXBP1


4-7

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016

OP-6

MOLECULARSIGNIFICANCEOFAUTOPHAGYINGAUCHERDISEASE

ÖzlemOral¹,ÖznurBayraktar²,AyselYüce³,SerapDökmeci⁴,DevrimGözüaçık²

¹|SabanciUniversity,NanotechnologyResearchandApplicationCenter²|SabanciUniversity,MolecularBiology,GeneticsandBioengineeringProgram

³|HacettepeUniversity,UnitofPediatricGastroenterology,HepatologyandNutrition⁴|HacettepeUniversity,DepartmentofMedicalBiology

Autophagyisalysosomal-dependentcatabolicpathwaycontributingtocellularhomeostasisbysequesteringcytosolic macromolecules in double or multimembrane vesicles and deliver them to lysosomes fordegradation.Gaucherdiseaseisthemostfrequentlysosomalstoragedisorder(LSD)causedbydeficiencyofacid-βglucosidaseandischaracterizedbytheaccumulationofglucosylceramideorothergycolipidsinvisceralorgans or central nervous system. Although the relevance of autophagy is shown in different LSDs, theunderlyingmolecularmechanisminGaucherdisease ispoorlyunderstood.Here,we investigatedmolecularsignificanceofautophagicpathwayinfibroblastscellsobtainedfromGaucherpatientshomozygousforL296Vmutation, aswell as for themost commonmutations,N370S, L444P, andD409H.Weobserved significantattenuation in theexpressionofkeyautophagy-relatedgenes (BECN1,ATG5andLC3)andaccumulationoftheirproteins inmutantcells.Wefoundthatdecreaseabilityofautophagosomestofusewith lysosomesisassociated with elevated lysosomal pH and reduced lysosomal enzyme activity. Analysis of proteasomaldegradationmachinery showeddecreasedproteolytic activity of proteasome,which consequently leads toincreased susceptibility to cell death. Our data indicate that both autophagic pathway and ubiquitin-proteasome system are affected bymulfunctional lysosomes andmay underlie themechanism of clinicalseverityofGaucherpatients.(ThisprojectissupportedbyTUBITAK-3501-NationalYoungResearchersCarreerDevelopment Program, Project No: 112T130).

THIS PROJECT IS SUPPORTED BY TUBITAK-3501-NATIONAL YOUNG RESEARCHERS CARREER DEVELOPMENTPROGRAM,PROJECTNO:112T130

Keywords:Autophagy,gaucherdisease,mutantfibroblasts


4-7

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016

OP-7

PROTEASOMEINHIBITORSTRIGGERNEUROPATHYBYDESTABILIZINGCYTOSKELETON

GülceSarı-Kaplan¹'²,SravaniMusunuri³,GrzegorzWicher⁴,TobiasJung⁵,JiaMi³,HüsniyeHacıoğlu-Bay6,JonasBergquist³,TilmanGrune⁵,BetülKarademir¹

¹|DepartmentofBiochemistry,MedicineFaculty/GeneticandMetabolicDiseasesResearchandInvestigationCenter,MarmaraUniversity,Istanbul,Turkey

²|DepartmentofGeneticsandBioengineering,EngineeringFaculty,OkanUniversity,Istanbul,Turkey³|DepartmentofChemistry-BMC,AnalyticalChemistry,UppsalaUniversity,Uppsala,Sweden

⁴|DepartmentofImmunology,GeneticsandPathology,Neuro-Oncology,UppsalaUniversity,Uppsala,Sweden

⁵|DepartmentofMolecularToxicology,GermanInstituteofHumanNutritionPotsdam-Rehbrücke,Germany6|DepartmentofAnatomy,MedicineFaculty,MarmaraUniversity,Istanbul,Turkey

Theproteasomal systemcontrolsdifferentcellularprocessesvia turnoverofmanycrucial cellularproteins.Therefore, proteasomal system is the target of treatment strategies for several diseases, including cancer.Since the approval of clinical usage of the first proteasome inhibitor, bortezomib, by FDA, proteasomeinhibitors have been used in the clinic for the treatment of different cancers. As many otherchemotherapeutic agents, peripheral neuropathy is one of themost abundant side effects of proteasomeinhibitors andmain reason for treatment limitations. Aim of the presentworkwas tracing the reasons ofperipheral neuropathy triggered by proteasome inhibitor bortezomib.With this purpose inmind, we alsocomparedbortezomibwithasecondgenerationproteasomeinhibitorcarfilzomib,whichisknowntobelesstoxicforneuroncells.Asabeginningstep,wecheckedtheinhibitoryeffectsofdrugsonproteasomeactivity.Bortezomibwas5-foldmoreeffectiveoninhibiting20Sproteasomeafter3htreatment,buttheninhibitoryeffects of drugs were similar. Then we performed proteomic analysis with nanoLC-MS/MS, and our datashowed significant changes in cytoskeletonmembers, especially in actin filament polymerization, vimentinand nestin expression, and also in HSP expression. We performed confocal imaging which also showedsignificant change in actin filament polymerization after 3 h drug treatment, andmicrotubule stabilizationafter24htreatment.Westernblottinganalysisshowedthat,whileARP2expression increasedabout2-foldafter3htreatmentinbothgroups,coronin1Cexpressionincreasedonlyinbortezomibgroup(1.2fold),andtransgelin expression change was greater in carfilzomib treated group after 3 h treatment.Coimmunoprecipitation of HSP70-β-actin (both directions) showed that β-actinmonomers interactedwithHSP70 chaperons after proteasome inhibitor treatment, and this interaction was higher in bortezomibtreated group. We can conclude that, the less toxic effect of carfilzomib may be related to actinpolymerizationstabilityandpotential.Keywords:bortezomib,carfilzomib,neuropathy


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016

OP-8

LONGNON-CODINGRNAS:POTENTIALREGULATORYPLAYERSOFAPOPTOSIS

UlviAhmadov¹,CanerBağcı¹,OsamaSweef¹,JensAllmer¹,AytenNalbant¹,BünyaminAkgül¹

¹|IzmirInstituteofTechnology

Apoptosis isa typeofProgrammedCellDeath (PCD)which isessential forcellularhomeostasisandproperdevelopment.Diseaseslikeautoimmunediseasesandcancerareassociatedwithaberrantapoptosis.Despitethewell-knownroleofcertainproteinsandmicroRNAsinapoptosis,thepotentialregulatoryroleoflongnon-coding RNAs (lncRNAs) is still unclear. In this study,we used two cancer therapeutics drugs, cisplatin anddoxorubicin,andtwoligands,FasmAbandTNFalpha,toidentifypathway-drugspecificand/orgloballncRNAsthat are differentially expressed in apoptotic HeLa cells. Induction of apoptosis was measured by FlowCytometry andwas further verified by FluorescenceMicroscopy andwestern blotting. Three replicates oftotalRNAsweredeep-sequencedusingtheIlluminaplatform.Treatmentwithcisplatin,doxorubicin,anti-Fasand TNF-alpha led to the differential expression of 1644, 506, 584 and 807, respectively. Total number ofdifferentially expressed lncRNAs that are common for all agents was ~250 (2-fold or higher, P < 0.01).Interestingly, a fraction of dysregulated lncRNAs appear to be antisense/sense to or located in the closeproximitytoprotein-codinggenesormiRNAsknowntobekeyregulatorsofapoptosis.Fourcandidateswereselected for validation by qPCR. GapmeR-mediated silencing of the candidates showed a relationshipbetween apoptosis and candidates. These results indicate that many lncRNAs are differentially expressedupon inductionofcelldeathvia treatmentwithcisplatin,doxorubicin,anti-FasandTNF-alpha inHeLacellsunder our experimental design. Mechanistic and further functional characterization of candidate lncRNAsmaygiveanimportantinsightintoapoptosisatthemolecularlevel. Keywords:apoptosis,longnon-codingRNA,deep-sequencing


4-7

May2

016

OP-9

HYPOXIAINDUCEDAPOPTOSISINMOUSENEUROBLASTOMACELLLINE

ElginTürközUluer¹,TunaÖnal¹,MehmetİbrahimTuğlu¹,CansuGörgün²,HafizeSedaVatansever¹'³

¹|DepartmentofHistologyandEmbryology,FacultyofMedicine,CelalBayarUniversity,Manisa,Turkey²|DepartmentofBiomedicalTechnologies,GraduateSchoolofNaturalandAppliedSciences,EgeUniversity,

Izmir,Turkey3|ExperimentalHealthScienceResearchCenter,NearEastUniversity,Nicosia,NorthCyprus

Object:Neuroblastoma(NB)isthemostcommonextracranialpediatricsolidtumor.Hypoxiaisawell-knownfeature of solid tumors and is a key prognostic factor for tumor progression and poor clinical outcome.Apoptosisisaprogressofprogrammedcelldeaththatoccursinresponsetodistinctsignalssuchashypoxia,excessiveoncogeneactivationorchemotherapeuticagents.Inthisstudyweaimedtoinvestigatethehypoxiceffectonapoptosisinneuroblastomacellline.Methods: Themouseneuroblastoma cell line (NA2B)was cultured inDMEMF-12 supplementedwith10%fetal calf serumand1%L-glutamine.Cellswere cultured in24wellplateanddivided into twogroups. Forhypoxic conditiongroup; cellswereexposed to3%O2,92%N2,5%CO2gasmixture tocreate3%hypoxiccondition for 36 hours in hypoxia chamber. For control group; cellswere incubated under normal cultureconditions in humidified atmosphere at 37°C in 5%CO2. After 36 hours incubation cells fixed in 4%paraformaldehyde and were stained with indirect immunoperoxidase technique in order to determinedistributions of cytochrome-c, bcl-2, Bax and caspase-3. DNA fragmentation was detected by terminaldeoxynucleotidyltransferase-biotinnickend-labelling (TUNEL)method.ResultsevaluatedwiththeOne-WayANOVAstatistical test.Results: Inhypoxicgroup,verystrong;caspase-3,strong;Baxandcytochrome-candmildtomoderate;Bcl-2 immunoreactivitiesweredetected. Immunoreactivitiesofcaspase-3,cytochrome-c,Bax and Bcl-2 were less in control group. TUNEL positive stained cells weremore in hypoxic groupwhencomparetocontrolgroup.Conclusion:Hypoxia is an important environmental factor that regulates cell differentiation, apoptosis andsurvival. In our study we demonstrated that hypoxia can induce apoptosis via the caspase activationaccompanying cytochrome c release with the increase of proapoptotic molecule Bax. High malignancypotential of NB can be correlated with resistance to apoptosis and hypoxia can be used to trigger theapoptosisoftheNB.Keywords:Apoptosis,hypoxia,neuroblastoma


4-7

May2

016

OP-10

PROTEASOMALSYSTEMANDHEATSHOCKPROTEINSINCANCERCELLDEATHMECHANISMS

BetülKarademir¹,AyşeMineYılmaz¹,ErgülMutluAltundağ¹,ErdiSözen¹,PerinurBozaykut¹,NesrinKartalÖzer¹,SemraKoçtürk²

¹|DepartmentofBiochemistry,MedicineFaculty/GeneticandMetabolicDiseasesResearchandInvestigationCenter,MarmaraUniversity,Istanbul,Turkey

²|DepartmentofBiochemistry,MedicineFaculty,DokuzEylulUniversity,Izmir,Turkey

Proteasomal degradation is crucial to prevent the accumulation of cellular damage. The removal of thedamageisarequiredprocessforhealthyorganismstokeeptheintegritywhile incancercellsthissituationmayinducedrugresistance.Regardingchemotherapy,degradationmechanismssuchasproteasomalsystemandautophagyhavebeen focused recentlyandproteasomal inhibition in cancer cellshavebeenshown toinduceautophagy.This inducedpathwaymaypreventthecancercells fromdeathorcancauseautophagiccelldeath.Therearemanypreclinicstudiestoimprovetheresultsandontheotherhandheatshockproteinsare accepted tobeprotectivewhichmaybringnewapproach. Theobjectof the studiesperformed inourlaboratorywastoconfirmtheroleofproteasomalsystemandheatshockresponse incancercelldeath. Inthisdirection,severalcancercelllineshavebeentestedfromdifferentaspectsofproteasomalactivity.MCF7andMDA-MB-231breastcancercellsweretreatedwithproteasomeinhibitorsbortezomibandcarfilzomibindifferentconcentrations.HCT116coloncancercelllineandHT22hippocampaltumorcelllinehavebeenusedto confirm the role of heat shock response related to proteasomal degradation. Results showed thatfollowing the treatment of breast cancer cells with bortezomib and carfilzomib, proteasome inhibitioninduced the autophagic protein degradation and apoptotic cell death. In this model, autophagic proteindegradation has been found to be a resistance against apoptotic cell death. In HCT116 cell line, heattreatment induced HSP70 protein expression which caused increased proteasomal degradation andautophagicproteindegradationanddecreasedapoptosis.Additionally,HT22cellsshowedthesameresultsindirectionwiththeothercelllines.Inthesecells,Nrf2pathwayhasbeenfoundtobeinvolvedintheactivationof proteasome activation and heat shock response. We can conclude that the activation of proteasomeactivity and regulation of the proteasomal system by heat shock proteins should be targeted in detail tohighlightthechemotherapyresistance.SupportedbyTUBITAKCOST-CM1001-110S281,TUBITAK212T156andTUBITAK115Z137.Keywords:proteasome,heatshockresponse,cancer,chemotherapy,resistance


4-7

May2

016

OP-11

THEROLEOFHYPOXIAONAPOPTOTICMARKERSINPRIMARYANDMETASTATICCANCERCELLLINES

TunaÖnal¹,ElginTürközUluer¹,CansuGörgün²,HafizeSedaVatansever¹'³

¹|DepartmentofHistologyandEmbryology,FacultyofMedicine,CelalBayarUniversity,Manisa,Turkey²|DepartmentofBiomedicalTechnologies,GraduateSchoolofNaturalandAppliedSciences,EgeUniversity,

Izmir,Turkey³|ExperimentalHealthScienceResearchCenter,NearEastUniversity,Nicosia,NorthCyprus

Aim: Cancer microenvironment is characterized by significantly lower oxygen concentration. Hypoxicconditionisacommonfeatureofsolidtumor,andisadirectstressthattriggersapoptosisinmanyhumancelltypesandpromotecellsurvival,motilityandtumorangiogenesisthatisessentialtotumor.Inthisstudy,weaimedtoinvestigatetheeffectofhypoxiaonapoptoticmarkersinprimaryandmetastaticcancercelllines.

MaterialsandMethods:Humanprimarycolon(Colo-320)andbreast(Mcf-7),humanmetastaticcolon(Colo-741)andbreast (M4A4)cancercell linewereused.Theywerecultured inRPMI-1640mediasupplementedwith10%FBS,1%L-glutamine,1%penicillinandstreptomycinuntil80%confluency.Alltypeofcellsdividedinto two groups; control group cultured with 5% CO2 and 37°C, hypoxia group were cultured in hypoxiachamberwhichhaveagasmixtureof5%CO2,3%O2and92%N2toprovidehypoxicconditionfor36h.Afterfixationwith4%paraformaldehyde,apoptoticcellsweredeterminedbyTUNELassay,distributionofCaspase-3,Bax,Bcl-2andCytochrome-cwereanalyzedusingindirectimmunohistochemistrymethod.ImmunostaningresultswereevaluatedasH-SCOREincomparisonwiththeOne-WayANOVAstatisticaltest.

Results: The number of TUNEL positive cells in Colo320 under hypoxic condition was higher than otherhypoxicandothergroups.Thestrong immunoreactivityofCaspase-3wasdetectedespeciallywasdetectedhypoxicconditionsofColo320cells.TheratioofBcl-2/Baxwashigher inprimarycancercells lines(Colo320andMcf-7).Themoderate/strongimmunoreactivityofCytochrome-cwasdetectedinallgroups.

Conclusion:Hypoxiaisimportantincoloncanceroccurrenceandprogression.Hypoxiaconditionpromotesapoptosisandinhibitsproliferationinvariouscancercellsinvitro.Ourresultsshowedthathypoxiatriggeredapoptoticpathwayespeciallyinprimarycoloncancercelllinehoweverbothcolonandbreastmetastaticcancercelllineswererescuedfromhypoxiainducedapoptosis.

Keywords:hypoxia,primary,metastatic,breast,colon,cancer,apoptosis


4-7

May2

016

OP-12

HL-60CELLLINEGENERATEDINVITROENVIRONMENTONANALYSISOFTHEBIOCHEMICALEFFECTSOFSORAFENIBANDLITHIUMCHLORIDE

AysunEkinci¹,CenapEkinci²,NurayYazıhan³,ElifKılıç⁴,AyhanBilir⁵,SafiyeKaya⁶

¹|DicleÜniv.TıpFak.BiyokimyaAnabilimDalı²|DicleÜniv.TıpFak.HistolojiveEmbriyolojiAnabilimDalı

³|AnkaraÜniversitesiTıpFak.FizyopatolojiBilimDalı⁴|BezmiAlemVakıfÜnv.TıpFak.BiyokimyaAnabilimDalı⁵|ZirveÜnv.TıpFak.HistolojiveEmbriyolojiAnabilimDalı

⁶|İ.Ü.CerrahpaşaTıpFak.BiyokimyaAnabilimDalı

Acute leukemia is characterisizedwithdecreasingmature cells andaggregationof leucocyteprecursors. InthisstudyweaimedtoinvestigatetheeffectsofSorafenibandLithiumChloride(LiCl)onHL-60humanAcutePromyelocytic Leukemia (APL) cell line. The HL-60 APL cell cultures are subjected to Sorafenib and LiClseparatelyand in combination for72hoursduringexperiments. Tumorcells are countedunderCellCountHemocytometerandtheapoptoseratiosare investigatedwith flowcytometrymethod.Thechanges in thecells in cell cultures are evaulated by transmission electron microscopy. To determine the possible themechanismofactionofthedrugs,thecaspase3actvities,phosphorylatedGSK-3β,phosphorylatedAKT,totalAKT, phosphorylated p38, total p38, phosphorylated ERK, total ERK, phosphorylated IκBα, total IκBα ,phosphorylated c-jun, total c-jun, phosphorylated STAT3 ve total STAT3 levels are investigatedwith ELISAmethod.SeparatelyandincombinationSorafenibandLiCldecreasedthenumberofcellsandincreasedtheapoptotic ratio in HL-60 APL cell line in a statistically significantmannerwhen comparedwith the controlgroup.

Keywords:HL-60,Sorafenib,LithiumChloride,Apoptosis,Leukemia


4-7

May2

016

OP-13

REGULATIONOFMIRNAONCOLONCANCERSTEMCELLS

FeyzanÖzdalKurt¹,RemziyeKendirci²,CananTürkoğlu¹,H.SedaVatansever²'³

¹|CelalBayarUniversity,FacultyofSciences&Letters,DepartmentofBiology,Manisa²|CelalBayarUniversity,SchoolofMedicine,HistologyandEmbryologyDepartment,Manisa³|NearEastUniversity,ExperimentalHealthScienceResearchCenter,Lefkosa,NorthCyprus

Colorectal cancer is one of themost commonly diagnosed and lethal cancersworldwide. It is amultistepprocessthatrequirestheaccumulationofgenetic/epigeneticaberrations.Thereareseveralissuesconcerningcolorectal carcinogenesis that remain unanswered, such as the cell of origin and the type of cells thatpropagate the tumorafter its initiation.Cancer stemcells (CSCs)have similarpropertieswithnormal stemcellsbut,theygrowrapidlyanddifferentiatetotumorcells.CSCsthereforeseemtohaveanimportantroleincancerrecurrence.Thebiologicalbehaviorofcancer,includingcarcinogenesisandfunctionalheterogeneity,canbeexplainedbytheCSChypothesis.TheregulationofCSCsatthemolecularlevelisnotwell-understood.MicroRNAs(miRNAs)areaclassofendogenousnon-codingRNAsthatplayanimportantroleintheregulationof several cellular, physiological, and developmental processes. Stem cell-specific miRNAs play importantroles in tumor initiation and development. Regulation of miRNA expression is controlling with severalproteins.Inthisstudy,thehumanprimarycoloncarcinomacell line,Colo320,RPMI-1640culturedin10%FCS,1%L-glutamine and 1% penicilin-streptomycine containing culture medium. Colon carcinoma stem cellscharacterized by CD133 surface proteinwere isolated fromprimer (Colo 320) colon carcinoma cell line bymagnetic-activatedcellsorting(MACS)technique.Magnetically labeledCD133+andunlabeledCD133−cellswere separated magnetically. CD133+ and CD133− cells were passaged after reaching 80% monolayerconfluency.Theywerethenfixedwith4%paraformaldydeanddistributionofanti-cyclinD1,anti-c-myc,anti-beta-catenin, anti-dicer, anti-drosha, anti-eIF2α and anti- eIF2C were investigated using indirectimmunoperoxidasestaining.Resultsshowthattheefficiencyoftheseparationsis30%,andtheseparationismore successful byMiniMACS column. Strong immunreactivity of cyclin D1was observed in primer coloncarcinomaCD133+andCD133-cells.Dicer,drosha, cyclinD1,c-myc,beta-catenin immunoreactivitieswerelessinCD133+cells.However,higherimmunoreactivitiesofeIF2αandeIF2CweredetectedinCD133+cells.AsaresultCD133+cellshavehigherexpressionofeIF2αandeIF2CthereforemiRNAexpressionsmaycontrolwith thesemolecules in CSCs. In addition, both CSCs and unlabeled Colo 320 cells have similar cell cyclecontrolling. Because of differences of regulation of miRNA expression between CD133+ and CD133- cells,treatmentprotocolshouldbedifferentintumors,whichhavecancerstemcells.

Keywords:Cancerstemcell,colon,cellcycle,miRNA


4-7

May2

016

OP-14

THEUSEOFKERATINOCYTESANDPRPINEXPERIMENTALBURNWOUNDMODELOFDIABETICRAT

HilalKabadayı¹,NavidH.Mansoub²,GülinnazErcan²,H.SedaVatansever¹'³

¹|CelalBayarUniversity,SchoolofMedicine,DepartmentofHistologyandEmbryology,Manisa,TURKEY²|EgeUniversity,SchoolofMedicine,DepartmentofBiochemistry,İzmirTURKEY

³|NearEastUniversity,ExperimentalHealthScienceResearchCenter,Nicosia,NorthCyprus

Object: Diabetes mellitus (DM) is a group of metabolic diseases characterized by hyperglycemia resultingfrom defects in insulin secretion, insulin action, or both. DM affects all three phases of wound healing(inflammation, proliferation and remodeling), leading to delayed closure and poor esthetic and functionalscarring.Akeratinocyteisthepredominantcelltypeintheepidermis.Theprimaryfunctionofkeratinocytesis the formation of a barrier against environmental damage. Besides, plateletrich plasma represent a keysourceof cytokines and growth factors extensively used for tissue regeneration;woundhealingand tissuerepair.In this studyweaimed to investigate the effects of keratinocyteswhichwasdifferentiated from ratadiposestemcellsandPRPinexperimentalburnwoundmodelofdiabeticrat(1,2,3).Material and Method: Male Sprague–Dawley rats were injected intraperitoneally with 60 mg / kgstreptozocin and after 4weeksDM statuswas defined as blood glucose levels higher than 280mg/dl. RatADSCs were isolated from retroperitoneal adipose tissue and washed with 5% penicillin-streptomycincontainingphosphatebufferedsaline(PBS).Briefly,theyweremincedintosmallpieces,treatedwith0.075%collagenasewithα-MEM(includes15%FCS,1%L-glutamin,1%penicillin-streptomycin)andincubatedat370C, 5%CO2 for 30minutes. The cellswere cultureduntil 80% confluency and thenwere differentiated tokeratinocyteusing0.5nMbonemorphogeneticprotein-4onmatrigel coatedcultureplate.Woundhealingratmodelwasobtainedwith100 °Cheated7mmdiametercopper thatapplied to thebackof rats for45seconds.Theywere then separatedasuntreated, keratinocyte,PRP,andkeratinocyte+PRPappliedgroups.Aftertransferringthekeratinocytes,tissuesampleswerecollectedonday3rd,7th,10thand14th.Theywerefixedwith10% formalin solution.Routineparaffinembeddingprocessesweredone. Sectionswere stainedbothH-Eandindirect-immunohistochemicalprotocolsusedforanalysinganti-EGF,anti-FGF2,anti-VEGF,anti-MCP1,anti-collagen1,anti-cytokeratin8,anti-cytokeratin14,anti-TGFβ-1,anti-PDGFimmunoreactivity.Results: Cellswere positively stainedwith early and late Keratinocyte differentiationmarker cytokeratin-8andcytokeratin-14,respectively.Cytokeratin-8immunoreactivitywasfoundmorepositivethancytokeratin-14. Histochemical analyses showed that, ADSC derived keratinocyte accelerate the epithelialization andimprove wound healing, but keratinocyte + PRP treatment were presented greatest improvement. Inaddition,molecularmarkersforprogressionofwoundhealingweremoredetectableinthisgroupaccordingtoothers.Conclusion:AlthoughapplyingADSCderivedkeratinocytestriggerthehealing,bestdevelopmentseenwhenkeratinocytesusedwithPRP.ThisstudyisabasicattempttounderstandthephysiologyofkeratinocytesandPRPbeforeusingthemforthefurtherstudies.

Keywords:woundhealing,stemcells,keratinocyte,diabetesmellitus


4-7

May2

016

OP-15

PROTECTIVEEFFECTSOFADIPOSE-DERIVEDMESENCHYMALSTEMCELLSONOVARIANISCHEMIA-REPERFUSIONINJURYINRATS

BurcuKasap²,ŞükrüKasap³,SedaVatansever¹'⁴,RemziyeKendirci¹,OsmanYılmaz⁵,MeryemÇalışır⁵,ÜmmühaniÖzelTürkçü⁶,MelikeNurAkın²,ErenAkbaba²,NilgünÖztürkTurhan²

¹|DepartmentofHistology-Embryology,SchoolofMedicine,CelalBayarUniversity,Manisa,Turkey²|DepartmentofObstetricsandGynecology,SchoolofMedicine,MuğlaSıtkıKoçmanUniversity,Mugla,

Turkey.³|DepartmentofPlastic,ReconstructiveandAestheticSurgery,SchoolofMedicine,MuğlaSıtkıKoçman

University,Mugla,Turkey4|ExperimentalHealthScienceResearchCenter,NearEastUniversity,Nicoisa,NorthCyprus

5|DepartmentofLaboratoryAnimalScience,SchoolofMedicine,DokuzEylulUniversity,İzmir,Turkey⁶|DepartmentofMedicalBiochemistry,SchoolofMedicine,MuğlaSıtkıKoçmanUniversity,Mugla,Turkey

Object:We aimed to investigate the immunohistological and biochemical effects of AdiposeDerived StemCells(ADSC)onovarianischemia-reperfusion(I/R)modelinrats.

Material andMethod: Twenty-five female, virgin adultWistar albino ratsweredivided into four groups asGroup 1 (Sham), Group 2 (ADSC), Group 3 (Bilateral torsion+detorsion), Group 4 (Bilateraltorsion+detorsion+ADSC).Laparotomieswereappliedtoallgroups,adnexaldetorsionwasappliedtoGroups3and4for6hperiod.After6-hperiodoftorsion/detorsionprocedures,bilateralovarieswereexposedand1x105cell/mL.ADSCwere seperatelyadministeredwithamicroinjector inavolumeof20µlPBSmedia inGroup 4. After 6 hour period over laparatomy, ADSC were administered to bilateral ovaries in Group 2.Laparotomieswereperformedinallanimalsaftera7-dayrecoveryperiodandbilateralovarieswereremovedandbloodwastakenfrominferiorvenacava.For immunohistologicalanalysis,TUNEL, iNOS,Caspase-3andBrdustainingweredeterminedinexperimentalgroups.Advancedproteinoxidationproteinproducts(AOPP)and total sulfhydryl (TSH) levels and superoxide dismutase (SOD ) activity were measured in serum andovariantissue.

Results: Serum AOPP levels as protein oxidation marker were significantly increased in Group 3 whencompared to Group 1 and Group 2 (p<0.05), whereas tissue and serum AOPP levels in Group 4 weresignificantlydecreasedcomparedtoGroup3(p<0.05).TissueSODactivityinGroup3and4weresignificantlydecreased compared to sham groups (p<0.05). iNOS wasmore strengthened via torsion and detorsion inGroup3,butdiminishedafterADSCimplementationinGroup4.ADSCtreatmentaftertorsion-detorsionhaveledtoadecreaseinapoptoticcellcountandCaspase-3immunoreactivity.ThesechangeswereallconsistentwithiNOSimmunoreactivityandTUNELstaining.

Conclusion:WeconcludedthatADSCmighthaveprotectiveeffectsonovarianI/Rmodel.

Keywords:KeyWords:AdiposeDerivedStemCell,Ischemia,Ovarian,Rat,Torsion


4-7

May2

016

OP-16

INVESTIGATIONOFANTICARCINOGENICEFFECTOFPOLYPHENOLCOMBINATIONSASADRUGONMCF-7ANDMCF-10ACELLLINE

MehmetFatihSeyhan¹,HülyaYılmaz-Aydoğan¹,AyçaDiren¹,ÖzlemTimirci-Kahraman¹,OğuzÖztürk¹

¹|IstanbulUniversity,InstituteofExperimentalMedicine,DepartmentofMolecularMedicine

Object: Recently, it has been showed that flavonoids have anticarcinogenic features. In our previousresearches,wedetectedthecellviabilityandcytotoxiceffectofmorethan30flavonoidsandphenolicacidsin breast cancer. In present study, we aimed that investigating the interactions and synergistic effects ofsomeflavonoidsandphenolicacidsuponMCF-7breastcancerandMCF-10Anormalepithelialcelllines.

MaterialandMethods:Forthispurpose,wedesignedartificialdrugcocktailsbasedontheseIC50valuesofthem, which contains these substances such as several flavonoid cocktails. Then, we performed the cellproliferationassaybyusingWST-1reagent,analysisofapoptosisbyusingAnnexinV-PIassayandcellcycleassaybyusingMuseCellAnalyzer.Afterdeterminingthemosteffectivedosesofdrugcocktail,weperformedthewholegenomemicroarrayanalysisandreal-timePCRexperimentcarriedouttovalidatethemicroarraydata.

Results:Inconsequenceofmicroarraydata,%75offlavonoidcocktailisthemosteffectivedosetoMCF-7andMCF-10Acelllines.InMCF-7breastcancercellline,thisdoseleadtomorethan60celldivisionrelatedgenesincrucialpointslikecellcycle,histones,centomereandkinetochore,gotdownregulated.Inaddition,morethan 30 DNA repair related geneswere down regulated and various breast cancer associated geneswereaffected. In contrast, there isno sucha similareffectuponMCF-10Abreastepithelial cell line. Ithasbeenshowedthatadesignedflavonoidcocktailpreventsthecelldivisionofbreastcellsinmanypathways.

Conclusion:Inthismanner,weconsiderthatflavonoidscanbeuseasananticancerdruginfurtherresearches.

Keywords:Flavonoids,phenolicacid,breastcancer,wholegenomexpression,cellproliferation


4-7

May2

016

OP-17

EFFECTOFTEAPOLYPHENOLSONPHASEIANDPHASEIIENZYMEACTIVITIESANDAPOPTOTICCELLDEATHMECHANISMINBREASTCANCERCELLS

AyşeMineYılmaz¹'²,ErgülMutluAltundağ¹'²,BetülKarademir¹'²,SemraKoçtürk²'³,YavuzTaga¹'²,A.SühaYalçın¹'²

¹|DepartmentofBiochemistry,SchoolofMedicine,MarmaraUniversity,Maltepe,34854Istanbul,Turkey.²|GeneticandMethabolicDiseaseResearchCenter,MarmaraUniversity,Maltepe,34854Istanbul,Turkey.³|DepartmentofBiochemistry,SchoolofMedicine,DokuzEylülUniversity,Inciralti,35340Izmir,Turkey.

Object:Inthisstudytheanti-proliferativeeffectofgreenandblackteapolyphenolsonbreastcancercelllineswereinvestigated.TheeffectsofpolyphenolextractsonphaseIandphaseIIenzymeswerealsoevaluated.Finally,theeffectsofthesepolyphenolsonapoptoticcelldeathmechanismswereinvestigated.

Materials andMethods: Polyphenolswere extracted from tea samples and obtained in powder formwithspray-drying. Extracts were then applied to breast cancer cell lines (MCF-7 and MDA MB-231) and theireffectsoncellviabilityandapoptoticmechanismswereinvestigatedbyflowcytometry.Additionallyquinonereductase(QR),glutathionetransferase(GST)activitiesweredetermined.

Results:InMCF-7cells,teapolyphenolsincreasedapoptosis(%17.7)andnecrosis(%36.5)valuessignificantly(p<0.05)whencomparedtocontrolgroup(%4.67and%4.97).ThiseffectwasnotobservedinMDAMB-231cells. Tea polyphenols increased green tea group quinone reductase activity (26.5 U/ml) in MCF-7 cellscompared to the control group (17.2 U/ml). For MDA MB-231 cells, almost no change in activity wasobserved.Quinonereductaseandglutathionetransferaseactiviteswereincreasedsignificantly(p<0.001)inMCF-7cellswhencomparedtothecontrolgroup.Glutathionetransferaseactivitywasincreasedonlyinblacktea group in MCF-7 cells, whereas it was decreased in MDA MB-231cells after both tea polyphenolsapplication.

Conclusion:Accordingtoourresults,polyphenolsappeartoeffectdifferentpathwaysdependingonthemolecularpropertiesofbreastcancercelllines.MCF-7cellsweremoresensitivetotheeffectsofpolyphenolsthanMDAMB-231cells.ApoptoticcelldeathwasinducedinMCF-7cells,whereasinMDAMB-231cellsotherdeathpathwayswereactivated.

Keywords:Breastcancer,Polyphenols,Celldeath,Cellcycle,Enzymeactivities.


4-7

May2

016

OP-18

THEAPOPTOTICEFFECTOFPISTACIAVERANUTSKINEXTRACTONHUMANGASTRICCANCERCELLLINEHGC-27

AbdullahTuncayDemiryürek¹,EbruTemiz²,SerdarÖztuzcu²,AhmetSaracaloğlu¹,ŞenizDemiryürek³,BilgeŞener⁴,BeyhanCengiz5,MustafaUlaşlı²,CelaletdinCamcı6

¹|DepartmentofMedicalPharmacology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

²|DepartmentofMedicalBiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey³|DepartmentofPhysiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

⁴|DepartmentofPharmacognosy,FacultyofPharmacy,GaziUniversity,06330Ankara,Turkey5|DepartmentofMedicalGenetics,FacultyofMedicine,GaziUniversity,06560Ankara,Turkey

6|DivisionofMedicalOncology,DepartmentofInternalMedicine,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

Objective:Gastriccancer(GC) isamajorworld-widehealthproblem.It isthethirdleadingcauseofcancer-relateddeath.Themedianoverallsurvival is lessthanoneyear foradvancedGCpatients; thus, there isanurgentneedtodevelopnoveltherapyforGC.Bioactivenaturalproductsareagoodsourcefordevelopmentof novel cancer preventive and therapeutic drugs. In this study, we investigated the apoptotic activity ofwaterextractofthepistachio(PistaciaveraL.)nutskinonthehumanGCcelllineHGC-27.

Material andMethod:HGC-27 cellswere cultured inDMEMmedium supplementedwith10% fetal bovineserum.Whencells reached70%ofconfluence, theywere treatedwithwaterextractorsalineduring24h.ApoptoticcellswerequantifiedbyAnnexin-V/7AAD-positivestaining,usinganAnnexin-V-FITC/7AADkitfromBeckmanCoulter.For thecell cycleassay,BDCycletest™PlusDNAReagentkit (Biosciences,Germany)wasused.Apoptosisandcellcycleanalysiswereperformedbyusingflowcytometer(NAVIOSBeckmanCoulter,USA).Forgeneexpressionstudy,mRNAwas isolatedfromcellsbyusingmiRNeasyMiniKit (QiagenGmbH,Germany). Then cDNAwas producedwith the Ipsogen RT Kit (QiagenGmbH). qRT-PCRwas performed byRotorGene6000(QiagenGmbH,Hilden,Germany).

Results: Apoptosis of theHGC-27 cellswas stimulatedwithwater extract (8.6%)when compared to saline(0.6%).Proliferationwasfoundtobestimulatedincellcycleassay(waterextract:G0/G157.0%,G20.0%,andS 43.0%; saline: G0/G1 45.1%, G2 0.4%, and S 54.5%). NFKB (2.5±0.4 fold), p27 (0.8±0.2 fold), and p53(4.1±2.3fold)geneexpressionsweresignificantlymodifiedwithwaterextracttreatment.

Conclusion: Our results showed that water extract of the Pistacia vera nut skin exhibited its anti-tumoractivity against HGC-27 cells through promoting apoptosis, inducing proliferation, and elevated p53 geneexpression.ThisstudywassupportedbyaGaziantepUniversityproject(TF.13.10).

Keywords:Apoptosis,cellcycle,flowcytometer,gastriccancer,geneexpression


4-7

May2

016

OP-19

RESVERATROLCANEFFECTCELLFATEDIFFERENTLYRELATEDWITHP53MUTATIONANDSIRT1ACTIVITYLEVELOFHCT116COLONCARCINOMACELLS

GüneşÖzen1,2,BelginSertSerdar1,HalilAteş3,SemraKoçtürk2

¹|DokuzEylülUniversity,InstituteofHealthSciences,DepartmentofMolecularMedicine,Inciraltı,Izmir²|DokuzEylülUniversity,FacultyofMedicine,DepartmentofBiochemistry,Inciraltı,Izmir³|DokuzEylülUniversity,FacultyofMedicine,DepartmentofHematology,Inciraltı,Izmir

Introduction: Resveratrol is a natural polyphenol synthesized by more than 70 plant species. Anti-carcinogenic effects of Resveratrol have been proved in a variety of cancer cells. However recent studiesemphasized that Resveratrol has an interaction with Sirtuin1 (SIRT1), a member of histone deacetylaseenzymes that having a role on critical genes and deacetylation of p53 in carcinoma. Considering theinteractionofResveratrolwithSIRT1activity,theinfluenceofResveratrolongeneexpressionsanditsrelationwithp53mutationisgaininggreatimportance.Aim:To investigate theeffectsofResveratrol inSIRT1 inhibitedanduninhibitedconditionsoncellviability,apoptotic cell death levels and fold changes of some proliferative or anti-proliferative gene expressions,whichmayhave importanteffectson colon carcinomaprogression, inp53(+/+) andp53(-/-)HCT116 coloncarcinomacells.MaterialandMethod: IC50dosesofresveratrolweredeterminedbyWST-1assay(Roche©).ThecelldeathratiosintreatmentsofResveratrolandSirtinol(aSIRT1inhbitor)weredeterminedbyAnnexin-V-FITC/PIassay(BD Pharmingen©) for flow cytometry (BD-FACS Canto II™). The activity of SIRT1 was evaluated byfluorometric activity assay (Enzo Life Sciences©). The changes of some the proliferative (CCND1, FRA1,PPARD, EGFR, BIRC5, PCNA, MCL1, STAT3, FOS, JUN) and anti-proliferative (P27, ATF4, TRAIL, PUMA,GADD45A,RB1,FASLG,TNF,SOCS3,STAT1)geneexpressionswereevaluatedinallgroupsasfoldchangesbyusingBio-RadCFXConnect™RealTimePCRDetectionSystem.AlldatawerestatisticallyanalysedbyStudent’sttestusingGraphpadPrism5.04software.Results: IC50levelofResveratrolwasfoundas54µMinHCT116p53(+/+)and31µMinHCT116p53(-/-)celllinesfor48hour.Accordingtoviabilitydata,wedecidedtouse60µMResveratrolconcentrationforbothofthecell lines.SIRT1 inhibitiondosageandtimingofSirtinolwerefoundas80µMand24hourrespectively.Resveratroltreatment(60µM)causes27.6%apoptoticcelldeathinSIRT1uninhibitedconditionbutitcauses33.4%apoptoticcelldeath inHCT116p53(+/+)cell linewithSIRT1 inhibitedcondition.Thesametreatmentconditions cause 17.9% and 18.2% apoptotic cell death for HCT116p53(-/-) cell line respectively. GeneexpressionresultsrevealedthatexceptforSurvivinandFOSgenes, inSIRT1 inhibitedconditionResveratrolcauses significant (p<0.05) increase of the proliferative genes in HCT116p53(+/+) cell line. However allproliferative gene expressions were found higher significantly (p<0.05) in HCT116p53(-/-) cell line for thesametreatments.Ontheotherhand,SIRT1 inhibitedconditioncausessignificant (p<0.05) increaseofanti-proliferative genes except for RB1 and SOCS3 in HCT116p53(-/-) cell line. The same conditionmakes alsosignificantrise(p<0.05)ofanti-proliferativegenesinHCT116p53(+/+)celllineexceptforRB1andGADD45A.Conclusion:ThisresearchhasrevealedthatResveratrol(60µM)causesdecreaseincellviabilityandincreaseinapoptoticcelldeathinHCT116p53(+/+)andHCT116p53(-/-)celllines.Inaddition,ourresearchprovedforthefirsttimethattheeffectsofResveratrolcouldchangeaccordingtop53mutationandSIRT1activitylevelofHCT116coloncarcinomacells.Weproposed thatResveratrolmight showproliferativeand/orapoptoticeffectsrelatedwithp53mutationandSIRT1activityofcoloncancercells.Consequently,wepredictedthatunconscious usage of Resveratrol in colon cancer patient might cause adverse effects. This study wassupportedwithScientificResearchProjectsCoordinationUnitofDokuzEylulUniversitywith2013.KB.SAG.055projectnumber.Keywords:Resveratrol,SIRT1,p53,ColonCancer,SignalTransductionPathways


4-7

May2

016

OP-20

ANTICANCERACTIVITIESOFDCM-MEOHEXTRACTSOFSALVIAFRIGIDAONLUNG,BREASTANDPROSTATECANCERS

ÖnderYumrutaş¹,İbrahimBozgeyik¹,SerdarÖztuzcu²,EsraBozgeyik²,MustafaPehlivan³,M.ÖzgürÇevik⁴,MünevverSökmen⁵,EbruTemiz²,PınarYumrutaş6,AhmetArslan²

¹|UniversityofAdiyaman,FacultyofMedicine,DepartmentofMedicalBiology,Adiyaman,Turkey²|UniversityofGaziantep,FacultyofMedicine,DepartmentofMedicalBiology,Gaziantep,Turkey³|UniversityofGaziantep,FacultyofSciencesandArts,DepartmentofBiology,Gaziantep,Turkey

⁴|UniversityofAdiyaman,FacultyofMedicine,DepartmentofMedicalGenetics,Adiyaman,Turkey⁵|KaradenizTechnicalUniversity,FacultyofScience,DepartmentofChemistryTrabzon,Turkey

6|UniversityofGaziantep,FacultyofMedicine,DepartmentofRespiratoryBiology,Gaziantep,Turkey

Object: The use of medicinal plants and their derivatives in the treatment of various disease is rapidlyincreasing.Salviafrigidaisamedicinalplantthatbelongstosalviagenus.Althoughithasbeenusedforalongtimedueitsmedicalactivities,itsanticanceractivitiesislargelyunknownandremainselusive.Inthisstudy,ouraimwastoevaluatetheanticancereffectsofSalviafrigidainlung,breast,andprostatecancercelllines.

MaterialandMethod:MTTwasusedtodeterminethecellviability.Apoptosisinductionandcellcyclephasesofcellswereevaluatedbyflow-cytometricapproach.Also,effectsoftheextractsonDNAfragmentationwereevaluated.Cellularantioxidantactivitiesofextractsweretested.ImportantphytochemicalsweredeterminedbyusingaHPLCandGCMS.Lastly,expressionofgenesinvolvedintheapoptosis,DNArepairandanti-oxidantsystemweredeterminedbyahigh-throughputapproach.Findings:ActivedosesofDCMandMeOHextractswasnot inducedapoptosis inA549cells.Also,DCMextractwas shown tobe inducedapoptosis inDU-145cells,yetMeOHshowedpoorapoptoticeffect.Additionally,MeOHwas increasedthenumberofapoptoticcellswith19.2%ratio.Incellcycleanalysis,bothextractwasnoteffectiveinA549cells.InDU-145cells,whileaneuploidy S cells were reduced, aneuploidy G2 cells increased after DCM extract treatment.While DCMextract had no effect on cell cycle of MCF7 cells, MeOH induced significant changes. Also, both extractsshowedstrongantioxidantactivitiesinallcells.Phenolicacidsofcatechin,syringicacid,epicatechin,routineand benzoic acid were determined by HPLC. In GCMS analysis, 28 phytochemicals in DCM and 20phytochemicals. Lastly, significant changes were determined in gene expression analysis after extracttreatments.

Conclusion:OurfindingssuggestthatSalviafrigidacanbenovelnaturalanticanceragent.Keywords:Anticancer,Apoptosis,Cancer,MedicinalPlants,Salviafrigida


4-7

May2

016

OP-21

THEEFFECTOFPROPOLISANDCAFFEICASIDPHENYLESTER(CAPE)INPANCREATICCANCER

AyferKarlıtepe¹,NerminKahraman²,BülentÖzpolat²,GülinnazErcan¹

¹|EgeUniversity²|TheUniversityofTexas

Introduction:Pancreaticcancer(PaCa)isoneofthemostlethalhumancancerswitha5-yearsurvivalrateof3–5 % (approximately 6 months survive). The most important features of PaCa is its early invasion andmetastasis,resistancetochemotherapyandradiotherapyandaggressivetumorprogression.Propolisanditscompoundscouldbepotentiallyusefulaschemotherapeuticorchemopreventiveanticancerdrugs.Bioactivecomponents from the propolis have been extensively explored to possess anticancer activity. Caffeic acidphenethyl ester (CAPE), a naturally occurring compound isolated from the extract of propolis with well-knownantioxidantactivity,hasbeen reportedasan inhibitorof certainenzymeactivities suchasxanthineoxidaseandcyclooxygenaseaswellastranscriptionalfactorNF-𝜅Bactivation.Material&Method:Cell Culture: PANC-1 cells were cultured in DMEM containing 10 % FBS, 2 mM L-glutamine, %1penicilin/streptomycin.WST1:SensitivityofthePanc1celllinetoincreaseddosesofpropolisandCAPEwasdeterminedbyculturing5×104cells/mlfor24and48h,incubatedwithWST1.ClonogenicAssay:TheeffectivenessofpropolisandCAPEonthesurvivalandproliferationofPanc1cellswastestedbyclonogenicassay.ApoptoticAssessment:TheapoptoticactivityofpropolisandCAPEonPanc1cellswastestedbyAnnexinVandTUNELassays.Result:WeinvestigatedtheeffectsofpropolisandCAPE inpancreticcancercellsandfoundthattheybothinhibitpancreaticcancercellproliferationandinduceapoptosis,butpropolisisfoundtobemoreefficientincomparisontoCAPE.


4-7

May2

016

OP-22

ANTI-CANCEREFFICACYOFDEGUELINAGAINSTLUNGCANCERCELLSWITHANDWITHOUTDOCETAXEL

SerapÇelebi1,NinaGhanitabe1,HakanCengiz1,HalilAteş2,MehmetAliKoçdor3,AzizKaraoğlu2,MeralKaraman4,HilalKoçdor1,2

¹|DokuzEylulUniversity,InstituteofHealthSciences,DepartmentofMolecularMedicine,IzmirTurkey2|DokuzEylulUniversity,InstituteofOncology,Izmir,Turkey

3|DokuzEylulUniversityFacultyofMedicine,DeparmentofGeneralSurgery,Izmir,Turkey4|DokuzEylulUniversityFacultyofMedicine,DeparmentofLaboratoryAnimalScience,Izmir,TurkeyOBJECTIVES:58,7%ofeverynew100.000casesthatwerediagnosedtocancerinbetweentheyears2008and2012are lungcancersandbronchialcarcinomas.LungcancersaredividedintwomaingroupsasSmallCellLung Cancers(SCLC) and Non-Small Cell Lung Cancers(NSCLC). Despite the recent improvements of thetreatments, the response and remission rates observed on the patients are relativelynominal.Dosetaksel(DTX)isachemotherapeuticthathasananti-tumoractivityagainstvarioussolidtumors.The growing resistance againstDTX still continues to be the biggest obstacle for the treatment success ofNSCLCpatients.Deguelinisanaturalplantderivativerotenoidandhasanencouragingactivityagainstalotofhuman cancers. The comparison of the treatment activity of the separate and combined usage ofDeguelin,whichisapotentialchemotherapeuticagent,andDosetaxel,whichisusedinstandardtreatment,isaimedinthisstudy.MATERIAL-METHOD: The IC50 doses of dosetataxel and deguelin on the A549 and H1299 NSCLCcell linesweredeterminedviathecellvitalitytestsinourstudy.TheactiveconcentrationsdeterminedwereappliedtoNSCLC cell linesasdeugelin,dosetaxel and their combinations. The impactsof themedicineare studiedbyapplying flow cytometric analyzes(apoptosis,cell cycle),glutathione and reducted glutathione,colonyformation,migration and angiogenesis analyzes on the treated cells and measuring the Oxidative StressIndex(OSI).Statistical analyse program,Rstudio(v.0.98.501) and the R-script languagewere used to examinethedifferencesbetweentheagents.Thestates inwhichthep-valuewaslowerthan0.05wereacceptedasstatisticallymeaningful.RESULTS:We found thatDeguelinhaspro-apoptotic,anti-migratoryandcytotoxicpotentialon lungcancercells. Deguelin amplifiedDTX-related anti-cancer efficacy(increased apoptotic cell content and cytotoxicity,reduced migration). Also,Deguelin pre-treatment sensitized the cells DTX-treatment(reduced IC50values).Theseeffectswereremarkableinp53-mutantcells.CONCLUSION: Deguelin, solely, has anti-cancer potential on NSCLCcells.Both Deguelin pre-treatment andcombinantionwithstandartchemotherapeuticsresultinenhancedanti-cancerefficacy.


4-7

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016

OP-23

SYNERGISTICEFFECTSOFACHILLEABIEBERSTEINIIAND5-FUAGAINSTCOLORECTALCANCERCELLS

MehmetKadirErdoğan¹,CanAliAğca² , HakanAşkın²

¹|BingölUniversity²|AtatürkUniversity

ObjectColorectalcancer(CRC)isamajorreasonofcancer-relateddeath,approximately1.2millionnewcasesarereportedeachyearintheworldwideandmorethanhalfdiefromthedisease.5-Fluorouracil(5-FU)isthebackbone in the clinical treatment of advanced CRC.However, beside the antitumor effectmost toxicitiesattributedtothedrugandobservedseveresideeffects.Therefore,noveltreatmentsarestillneeded.5-FU-based chemotherapeutic regimens are established as a fundamental standart treatment for metastaticcolorectal cancer [1]. Antimicrobial, antioxidant, antiinflammatory, spasmolytic, antidiabetic, antiulcer,antitumor, cholereticandhepatoprotectiveactivity, andcytotoxiceffectsofdifferentAchillea specieshavebeen previously reported [2]. A. biebersteinii Afan. (Asteraceae) is a perennial herb, villose, stems erect,simpleorbranchedfromthebase;30–60cmhigh;leavesupto10cm;floweringperiod,April-May[3].Inthisstudy,antiproliferativeandapoptoticeffectsofA.biebersteiniiandit’scombinationwith5-FUwereanalyzedwithvariousmethods.MaterialandMethodHT-29colorectaladenocarcinomacelllinewereobtainedfromATCC.CellProliferationKit I (MTT) and Cell Death Detection Elisa Kit were purchased from Roche Diagnostics, Germany. Otherchemicalsandreagentswereobtained fromSigmaandMerck.CellviabilitywasdeterminedbyMTTassay.HT-29cellsweretreatedwiththedifferentconcentrationsofA.biebersteinii,5-FUandA.biebersteinii+5-FU.Cell DeathDetection Elisa Kitwas used according to themanufacturer’s protocol for detect the apoptoticeffect.pTEN,AKT,MAPK,mTOR,VEGFReceptor2,p53andβ-actingeneexpressionlevelsweremeasuredbyRT-PCR.WesternblotanalyzewereusedtodeterminepTEN,AKT,MAPK,mTOR,VEGFReceptor2,p53andβ-actinproteinlevels.Results Results are provided as the mean of independent experiments, each assay were performed intriplicate.ConclusionInvitrocytotoxicandapoptoticeffectofA.biebersteinii+5-FUshowedthatthiscombinationcanbeacandidateforcolorectalcancertreatment.Keywords:Achillea,apoptosis,5-FU,mTOR


4-7

May2

016

OP-24

ANTI-PROLIFERATIVEANDANTI-INVASIVEEFFECTSOFFERULICACIDINTTMEDULLARYTHYROIDCANCERCELLSINTERACTINGWITHURG4/URGCP

YavuzDodurga¹,CananEroğlu²,MücahitSeçme¹,LeventElmas¹,ÇığırBirayAvcı3,N.LaleŞatıroğlu-Tufan4

¹|PamukkaleUniversity,SchoolofMedicine,DepartmentofMedicalBiology²|NecmettinErbakanUniversity,SchoolofMedicine,DepartmentofMedicalBiology

³|EgeUniversity,SchoolofMedicine,DepartmentofMedicalBiology4|AnkaraUniversity,SchoolofMedicine,DepartmentofForensicMedicine

OBJECT:Ferulicacid(4-hydroxy-3-methoxycinnamicacid;FA),acommondietaryplantphenoliccompound,isabundant infruitsandvegetables.Theaimofpresentstudy isto investigatetheeffectsofFAoncellcycle,apoptosis,invasion,migration,andcolonyformationintheTTmedullarythyroidcancercellline.

MATERIAL ANDMETHOD: TT human thyroid cancer cell line treated with 50 µM - 1mM FA by solving inmediumfor24,48and72hconsideringatime-anddose-dependentmanner.TheeffectofFAoncellviabilitywas determined by using CellTiter-Glo Cell Viability Assay. Expression profiles of certain cell cycle andapoptosis genes such as URG4/ URGCP, CCND1, CDK4, CDK6, p53, PARP, PUMA, NOXA, BAX, BCL2, BID,CASP3, CASP9,MMP2,MMP9 and TIMP1were performed on real-time PCR according to the SYBR Greenprotocol. Effects of FA in TT cells on invasion, colony formation and cell migration were determined bymatrigel chamber, wound-healing and colony formation assay, respectively. Statistical analysis wereperformedwithRT2ProfilesPCRArrayDataAnalysiswhichisassessedstatisticallyusingtheStudent’sttest.

RESULTS: IC50 dose of FA in the TT cells was detected as 150 μM. It was determined that FA caused adecrease in the expression of URG4/URGCP, CCND1, CDK4, CDK6, BCL2, MMP2 and MMP9, a significantincrease intheexpressionofp53,PARP,PUMA,NOXA,BAX,BID,CASP3,CASP9andTIMP1genes inTTcellline. It was also found that FA in TT cells suppressed invasion, migration, and colony formation by usingmatrigelinvasionchamber,woundhealingandcolonyformationassay,respectively.

CONCLUSION: FA indicates anti-carcinogenesis activity by affecting cell cycle arrest, apoptosis, invasion,migration,andcolonyformationonTTcells.ItisnecessarytoconductfurtherstudiestodiscovertherapeuticeffectandthemolecularmechanismofFAonthyroidcancer.

Keywords:Ferulicacid,Thyroidcancer,URG4/URGCP,apoptoticandcellcyclegenes


4-7

May2

016

OP-25

RESVERATOLINDUCESAPOPTOSISTHROUGHOXIDATIVESTRESSINBIPHASICMALIGNANTPLEURALMESOTHELIOMACELLS

SaimeBatırel¹'²,ErgülMutluAltundağ¹'²,ElifKurt¹,NesrinKartalÖzer¹'²,HasanFevziBatırel³

¹|DepartmentofMedicalBiochemistry,FacultyofMedicine,MarmaraUniversity,Istanbul²|GeneticandMetabolicDiseasesResearchCenter(GEMHAM),MarmaraUniversity,Istanbul

³|DepartmentofThoracicSurgery,FacultyofMedicine,MarmaraUniversity,Istanbul

AIM: Malignant pleural mesothelioma (MPM) is an asbestos-related tumor arising from the mesothelialsurfaceofthepleuralcavitywithapoorprognosis.Biphasicmalignantpleuralmesothelioma(BMPM)istheoneofthemostresistantsubtypeofMPMtoradiationandchemotherapy.Resveratrol(RSV)isapromisingnatural compound in the treatment of cancer. Its potent anti-cancer activity on BMPM cells are shown inprevious studies.However theunderlyingmolecularmechanismsof thiseffect isunclear. Therefore in thisstudy we investigated the mechanism of the anti-proliferative effect of RSV in BMPM cellsMATERIALANDMETHOD:HumanBMPMcells(MSTO-211H)weretreatedwithRSVatconcentrationsfrom5μM to 150 μM for different exposure times (24h, 48h, 72 h) and then cell viability were measured. Toevaluateapoptosis,thecellsweretreatedwithRSVfor24hoursandthenannexinV-FITC-propidiumiodide(PI) double staining is performed. ROS production in the cells ismeasured by flow cytometry and proteinexpressionsofanti-oxidantenzymesaremeasured.Furthermore,sinceNFKBpathwayisakeyinpreventionofapoptosis,weexaminedtheproteinexpressionofit.RESULTS: RSV treatment reduced the cell viability in a dose and time dependent manner. Apoptosis wassignificantlyincreasedinRSV-treatedcells.IntracellularROSlevelswerenotchangedwithtreatmentbutRSVincreased SOD2 protein expression although it didn’t change GPX and SOD1 protein expressions.CONCLUSION:OurresultsrevealedthatRSVexhibitedsignificantanti-proliferativeeffectonBMPMcellsinadoseandtimedependentmannerbyinducingapoptosisandthisinductionofapoptosisinthecellsmayhaveoccurredviaincreasedROSproduction.AndthisoxidativestressmaybecounteractedwithincreasedSOD2level.ThesedatasuggestthatRSVmaybeanimportantsubstancetoinvestigateclinicallyforthetreatmentofBMPMwhichisaveryresistantsubtypeofMPMtotherapy.Keywords:Resveratrol,BiphasicMalignantPleuralMesothelioma,Apoptosis


4-7

May2

016

OP-26

EVALUATIONOFANTIPROLIFERATIVEACTIVITYANDANTIAPOPTOTICEFFECTOFTHYMBRASINTENISIISUBSP.ISAURICAEXTRACTSINHUMANCANCERCELLLINES

CeylanHepokur¹,SemaMısır¹,MehmetÇiçek²

¹|CumhuriyetUniversity²|PamukkaleUniversity

Object: Cancer is a pathological state that is a genetic and developmental process occurring due to theexcessiveproliferationofthecellsandthelossoftheirapoptosisfunctions.Sideeffectsanddrugresistancecauseproblemsintheusageofsyntheticdrugs.Therefore,researcherstrytoelucidatenewdrugcandidatecompounds from bioactive natural products to lessen-prevent side effects and resistance. Traditionalmedicine from plants leads a pathway for this purpose. Thymbras also display antioxidant activities, anti-tumourandantimicrobialactivities.Severalplantspecieshavebeenusedforpharmaceuticalpurposesandyetmanyplantsneedtobestudiedinmoleculardetail.Thymbrashaverichfloraandattractedresearchersfor elucidating their composition. In the present study, we aimed to investigate antiapoptotic effect,anticancer,antioxidantactivityofThymbrasintenisiisubsp.isauricaextracts.MaterialandMethod:AntiproliferativeActivityThebreastcancercellline(MCF-7),humanosteosarcomacellline(MG-63),mousefibroblastcellline(L929)weretreatedwiththedifferentconcentrations(5,10,20,40,80, 160 µg/mL) of Thymbra sintenisii subsp. isaurica extracts in 24 and 48 hours and the analysis ofcytotoxicitywastestedwithXTT.TheIC50valueswerecalculated.AntioxidantActivitiesAntioxidantactivitiesof Thymbra sintenisii subsp. İsaurica extractswas investigatedbymeansof free radical scavenging activity(DPPHassay) and total phenolic compounds.AnnexinVAnalysisAntiapoptotic Effect of Thymbra sintenisiisubsp.İsauricaextractswasevaluatedintermsofapoptosisusingflowcytometry.Results: As a result of the study, it has been found that rich in polyphenolic compounds, high radicalscavengingactivityandanticanceractivityofThymbrasintenisiisubsp.İsauricaextracts.Conclusion:Thymbrasintenisiisubsp. İsauricaextractsshowedpotentanticanceractivityagainstcancercelllines.

Keywords:Thymbrasintenisiisubsp.İsaurica,Apoptosis,Cancer,Polyphenol


4-7

May2

016

OP-27

BACTERIALHEATSHOCKPROTEINGROELTRIGGERSHUMANPRIMARYTCELLAPOPTOSIS

AytenNalbant¹,BünyaminAkgül¹

¹|MolecularImmunologyandGeneRegulationLaboratory,DepartmentofMolecularBiologyandGenetics,FacultyofScience,IzmirInstituteofTechnology

Objectives: Modulation of apoptosis could be a critical factor in defining the outcome of an infection bybacterial virulence factors.Aggregatibacter actinomycetemcomitans’ Hsp is a 64-kDa GroEL-protein whichhasbeenshowntoinfluencethehostandimmunesystemcellsbutapoptoticcapacityofGroELproteinonTcellsisnotknownyet.ThepurposeofthepresentstudywastoinvestigatetheabilityofendogenousGroELproteinofAggregatibacteractinomycetemcomitanstoinducehumanTcellapoptosis.Methods:EndogenouslyexpressedGroELproteinwaspurifiedfromA.actinomycetemcomitans(ATCC29522)byATPaffinitychromatographyandelectroelutedfromSDS-PAGE.PurifiedGroELproteinwasconfirmedbywesternblotandLC-ESI-MS.LPSconcentrationinpurifiedGroELproteinwasanalyzedbyLALChromogenicEndpoint assay kit. Detoxi-Gel Endotoxin Removing Gel was used to remove LPS from purified samples.PurifiedGroELproteinwasusedasantigenatdifferentdoses.PBMCsfromhealthydonorswereculturedwithGroELfrom0-96hours.ApoptosisrelatedchangesinTcellsweremeasuredbyflowcytometryandwesternblot.Results:ThedatashowedthatphosphatidylserineexposureasanearlyapoptoticeventwasdosedependentinGroELtreatedTcells.ThekineticsofplasmamembranechangesofTcellswerealsotimedependent.GroELtreatedTcellswerepositiveforactivecaspase-3inadosedependentmanner.Additionally,therateofGroELinduced apoptosis was suppressed by the addition of general caspase inhibitor Z-VAD-FMK. Furthermore,cleavedcaspase-8bands(40/36kDaand23kDa)wereidentifiedinGroELrespondingcells.Conclusions:OveralldatapresentedinthisstudydemonstratedthatendogenousheatshockproteinGroELofA.actinomycetemcomitansmediatesTcellapoptosissuggestingaroleforHsp’stomodulateTcell immuneresponse.Acknowledgements:ThisworkwassupportedbyTUBITAK(Grant#106T417toDr.AytenNalbant).Keywords: T cells, Apoptosis, GroEL, Bacterial heat shock protein, Hsp60, Aggregatibacteractinomycetemcomitans


4-7

May2

016

OP-28

CANADV36INDUCE3T3-L1PREADIPOCYTESINTOMATUREADIPOCYTES?

TamerŞanlıdağ¹'²,SedaVatansever¹'²'³,SinemAkçalı¹,SevtapGökalp¹'⁴,MehtapKoçan¹ ,FerdiyeTaner¹'²'⁴

¹|CelalBayarUniversityFacultyofMedicine,DepartmentofMedicalMicrobiology,ManisaTurkey²|NearEastUniversity,ResearchCenterofExperimentalHealthSciences,Nicosia-NorthCyprus

³|NearEastUniversity,FacultyofMedicine,DepartmentofMedicalMicrobiology,Nicosia-NorthCyprus4|CelalBayarUniversityFacultyofMedicine,DepartmentofHistologyandEmbryology,ManisaTurkey

Aim: 3T3-L1 adipocytes originally derived from Swiss mouse embryo tissue have been fundamental inmetabolicdiseaseresearchfor30years.The3T3-L1systemhasbeenpivotalinadvancingtheunderstandingof basic cellularmechanisms associatedwith diabetes, obesity and related disorders. In this study, it wasaimed to investigate thedifferentiationofpreadipocytes infectedwithAdenovirus36usingmorphological,histochemicalandimmunohistochemicalmethods.Method:Followingtheremovalof3T3-L1preadipocytecelllinesfromstock,cellswereincubatedina3T3-L1preadipocyte liquidmedium(ZenBio, Inc,USA) for tendays.ThemediumwascomposedofDMEM,HEPES,Bovinecalfserumtogetherwithaselectionofantibiotics(i.e.,Penicillin,Streptomycin)andanantifungal(i.e.,AmphotericinB).Thepreadipocyteculturemediumwasreplacedevery2dayswithfreshmediumduringthe10dayincubationperiod.Thesubsequentformationof~80%ofconfluentcellsallowedthepassageofcellsinto twogroupsusing6-well plates. Following twodaysof incubation,oneof thepassaged cell lineswereinfectedwithAdv36(ATCCVR-1610)whiletheotherremaineduninfectedandservedasthecontrolcellline.Cellswerecollectedfromboththeinfectedandcontrolcelllinesat2,5,7,9,14and21daysandusedinphasecontrastmicroscopy,OilRedstainingandleptinimmunoreactivityanalyses.ResultPhasecontrastmicroscopyanalysisrevealedanincreaseinthenumberoffatvacuolesamongsttheAdv36infectedpreadipocyteswhencomparedwith theuninfectedcell line.Furthermore, thedifferentiationof the infectedpreadipocytes intomature adipocytes markedly increased particularly after the 7th day of infection. This observation wasconfirmed with the microscopy studies, together with Oil red staining and leptin distribution detectionmethods.Conclusion:Adv36infectioncantrigger3T3-L1preadipocytesintomatureadipocytesinadditiontoincreasingthenumberoflipidvacuoles.

Keywords:Adenovirus36,3T3-L1cell,preadipocyte,matureadipocyte


4-7

May2

016

OP-29

NUCLEOFECTIONEFFECTONEARLYAPOPTOTICCHANGESINHUMANNAIVECD4TCELLS

SeminayGüler1,BünyaminAkgül1,AytenNalbant1

¹|İzmirYüksekTeknolojiEnstitüsü,MolekülerBiyolojiveGenetikBölümüUrla,35430İzmir,Turkey

Objective: Naive CD4 T cells play amajor role inmediating immune response. These cells are difficult totransfectwithviralbasedmethodsbecauseof somedrawbacks.Theaimof this study is to investigate theeffectofnucleofection,anon-viraltransfectionmethod,onNaiveTcellapoptosis.

Methods: Blood form healthy donors is obtained with the permission of Dokuz Eylül Faculty ofMedicinenoninvasiveethicscommittee.Humanperipheralbloodmononuclearcells(PBMC)areisolatedwiththeficolldensitygradientcentrifugationNaiveCD4TcellareisolatedbyutilizingVariomax.Afterthepurityofthecellsaremeasured, theAMAXA4DNucleofectionsystem isusedonthesecells for the introductionofGFP.ThecellsarethenculturedinfullIMDMfor24and48h.Then,thecellswereanalyzedwithbothCD4,CD25,CD69markersforcellactivationandAnnexinVand7AADmarkersforcelldeathbyusingFlowCytometry.

Result: The rate of CD4 positive GFP positive cells is found 0.3% after 24h incubation and 35% after 48hincubation.Thecellsarealsoanalyzedwithactivationmarkers suchasCD25,CD69.Both the rateofCD25positive GFP positive cells and CD69 positive GFP positive cells are found 0% after 48h incubation . TheviabilityofthenucleofectedcellsaresearchedbyAnnexinVand7AAD.TherateofAnnexinVpositiveGFPpositive cells is found ~25% and the rate of 7AAD positive GFP positive cells is found ~8% after 48hincubation.

Conclusion:Inconlusion,Nucleofectionisaneasyandrapidmethodthatgivesshortelectricalpulsestocellmembraneandmakeholes in themembrane throughwhichnucleicacids canpass.Due to thechanges inphospholipidstructure,thecontinuityofcellfunctioncaneffectandcellscanundergoapoptosis.


4-7

May2

016

OP-30

INHIBITIONOFCERAMIDASESISANEWPOTENTIALTARGETFORLIVERCANCERTHERAPY

HaticeMehtapKutlu¹,DjananVejselova¹,GökhanKuş²

¹|DepartmentofBiology,FacultyofScience,AnadoluUniversity,YunusemreCampus,26470,Eskişehir,Turkey.

²|DepartmentofHealth,FacultyofOpenEducation,AnadoluUniversity,YunusemreCampus,26470,Eskişehir,Turkey.

Object:Human liverhepatocellularcarcinoma(HCC) isoneof themostcommonmalignancies in theworldwithanestimatedhalfamilliondeathsannuallyanditsincidenceisontheriseintheUSA,EuropeandAsia.HCC is highly resistance to chemotherapy. Ceranib-2 is a ceramidase inhibitor that has shown significantantitumoractivityinavarietyoftumorcells.Weintendedtoinvestigatewhetherceranib-2inhibits/inducescellproliferationandapoptosistheofhepatomacancercelllinesHepG2andSK1.Toevaluateifceranib-2hasan activity against liver cancer and with an aim to identify the altered cellular factors upon ceranib-2treatment.HumanHepG2andSK-1cancercelllineswasusedasamodelandcelldeathapproachwasutilizedtoelucidatethemolecularmechanismsunderlyingceranib-2’santitumoractivity.

MaterialandMethod:HepG2andSK1celllineswerepurchasedfromATCC.HepG2cellsweremaintainedinDulbecco’smodified Eagle’smediumand SK1 in cells EMEMmediumat 370C and5%CO2 in atmosphere.Cells growth inhibition was measured by MTT method and apoptosis was detected by flow cytometry.Ultrastructural changes in HepG2 and SK1 cellswere investigated under transmission electronmicroscopeandmorphologicalchangesunderconfocalmicroscope.

Results: Ceranib-2 remarkably inhibited the proliferation of HepG2 and SK1 cancer cells and inducedapoptosis in ceranib-2 treatedgroups.According toour transmissionelectronmicroscopy results ceranib-2altered the ultrastructure in both of SK-1 and HepG2 cells. In ceranib-2 treated and acridine orange andphalloidin stained cells themorphology of SK-1 andHepG2 cellswas altered indicated apoptotic changes.Flowcytometricanalysisunderliestheapoptoticactionofceranib-2bothinSK-1andHepG2cells.

Conclusion:Onthebasisorourfindingswesuggestthisagentforfurtherresearchforcancertherapy.

Keywords:Ceranib-2,livercancer,apoptosis.


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OP-31

THEEFFECTSOFBORCOMPOUNDSONINFLAMMATORYGENESINBREASTCANCER

KeremAltun²,BuketÖzen²,EfeSerinan¹,SafiyeAktaş¹,BurcuTepedelenErbaykent³,MehmetKorkmaz³ ,ZekiyeAltun¹

¹|DokuzEylulUniversity,InstituteofOncology,BasicOncologyDepartment,Izmir,TURKEY²|TAKEVCollege,Izmir,TURKEY

³|CelalBayarUniversity,SchoolofMedicine,MedicalBiologyDepartment,Manisa,TURKEY

Objective:Breastcanceristhemostcommonwomen’scancerintheworld.Borcompoundshaveanti-cancerproperties.Theaimofthisstudywastoevaluatetheeffectsofborcompoundssuchasboricacid(BA)anddisodiumpentaborate(DSP)inbreastcancercellsviaapoptosisandinflammatorygeneexpressions.Material andMethods: Triple negative (estrogen, progesterone and ERB2) and positive breast cancer celllines,MDA-MB-231 and HTB-20 cells, treated with BA(0-200uM) and DSP(0-7.5uM). Cell proliferation andapoptosis determined with using WST-1 and Flow cytometric Annexin-V/PI measurements. Inflammationrelated84geneexpressionswereevaluatedwithRT-PCRarray.Kruskal-WallisandMann-WithneyUtestandalsot-testwereusedforstaticallyanalysiswithusingSPSS15.0program.≥10foldandmorechangeswereacceptedasasignificantgeneexpressionlevel.Results: Both of BA (50uM) and DSP (5uM) decreased the cell viability by dose dependentmanner at 24hours’incubation.ApoptoticcelldeathwasalsoinducedmainlyinHTB-20triplepositivebreastcancercells.BA treated MDA-MB-231 cells showed that increased CCL15, CCR1, CSF2, CXCL11, IL-15 while decreasedCCL22,IL-7,OSM,TNSF10geneexpressions.DSPincreasedtheexpressionsofCSF1,CXCL3,IL10RA,IL16,IL27,LTB,MIF, TNFRS11Bbutdecreased theexpressionsof IL5 in same cells. InHTB-20 cells, BA reducedCCL1,CCL4,CCL5,CCR1,CD40LG,CXCL2,CXCL6,CXCR1,FASLG,IL27,IL5,NAMPT,OSM,TNFS13Bgeneexpressions.AIMP1,CCL13,CCL8,CXCL12,CXCL5,CXCR1,FASLG,IL10RA,IL13,IL1R1,IL7,TNSF10,TNSF4geneexpressionsincreasedwithDSPinthatcells.Conclusion:BAandDSPaffectedtheexpressionsofinflammationrelatedgenesinbreastcancercellsintheopposite directions. This study showed that BA and DSP have different effects on specially inflammationrelatedgenesbydirectedestrogen,progesteronereceptorsandHER2inbreastcancer.

Keywords:Boricacid,Disodiumpentaborate,breastcancer,inflammatorygenes,apoptosis


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016

OP-32

EVALUATIONOFTHEAPOPTOTICEFFECTSOFWORTMANNINANDTHALIDOMIDEONBREASTCANCERCELLLINES

MelikeÖzgül¹,ElginTürközUluer¹,GamzeTanrıöver²,Sevinçİnan¹

¹|ManisaCelalbayarUniversity,FacultyofMedicine,DepartmentofHistology&Embryology,Manisa,TURKEY

²|AkdenizUniversity,FacultyofMedicine,DepartmentofHistology&Embryology,Antalya,TURKEY

Object:Theaimof thisstudywasto investigatetheeffectsofPI3K inhibitorWortmanninandangiogenesisinhibitorThalidomideonintrinsicandextrinsicapoptoticpathwaysonbreastcancercelllineswhichhavelow(67NR)andhigh(4T1)metastaticpotentialusingTUNELandindirectimmunohistochemicaltechniques.MaterialandMethod:67NRand4T1breastcancerlineswereculturedinDMEM-F12,mediumcontaining5%FBS, 1%NEA, 1% L-glutamine and 1%penicillin/streptomycin. IC50 valueswere determined as 2,5 µM forWortmannin and 25 µM for Thalidomide by using MTT assay. Apoptotic cells were detected via TUNELmethodandTUNELindexwerecalculated.Anti-Caspase-3,anti-FasL,anti-Apaf-1,anti-Cytochrome-candanti-Bcl-2primaryantibodieswereperformedimmunohistochemicallyafter24hand48hdrugadministration.Themeanvaluesofthestainingintensities(mild,moderate,strongandverystrong)andpercentageofpositivelystainedcellswerecalculatedusingH-Score.Results: It was observed that TUNEL index had a statistically significant increase in the drug administeredgroupwhencompared to thecontrolgroups (p<0.05). ImmunoreactivitiesofCaspase3,FasLigand,Apaf-1andCytochromeCwere seenasmild,mild/moderate, strongandmoderate/strong in67NRcontrol group,respectively. While immunoreactivity of Caspase 3 was seen as moderate in 4T1 control group,immunoreactivitiesofFasLigand,Apaf-1andCytochromeCwereobservedasmoderate/stronginthisgroup.Statisticallysignificantincreasedimmunoreactivityscoresweredeterminedinthedrugadministeredgroupswhen compared to the control groups (p<0.05). The immunoreactivity of Bcl-2 was seen as strong in allgroupsandtherewasnostatisticallysignificantdifferenceforBcl-2immunoreactivity(p>0.05).Conclusion:ItwasconcludedthatWortmanninandThalidomidehadanimportantroleonbothintrinsicandextrinsicapoptoticsignallingpathwayson67NRand4T1breastcancercelllines.Asfutureexpectation,thesedrugs might be used therapeutically to control cancer development via apoptosis in addition to classicaltreatmentsoncancertreatment.

Keywords:BreastCancerCellLine,Wortmannin,Thalidomide,apoptosis.


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Ø POSTERPRESENTATIONS


4-7

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016

PP-1

ASCAFFOLDPROTEINISANINTERACTIONPARTNEROFATG5ANDANOVELREGULATOROFAUTOPHAGY

SeçilErbil¹,ÖzlemOral¹,GeraldineMitou¹,CenkKig¹,EmelDurmaz-Timuçin¹,EmineGüven-Maiorov²,FerahGülaçtı¹,GökçenGökçe¹,JörnDengjel3,OsmanUğurSezerman4,Devrim

Gözüaçık¹

¹|SABANCIUniversity,MolecularBiology,GeneticsandBioengineeringProgram,Orhanli-Tuzla,34956Istanbul,Turkey,²|KOCUniversity

³|FreiburgUniversity4|AcibademUniversity

Autophagy isabiologicalmechanismallowingrecyclingof long-livedproteins,abnormalproteinaggregatesand damaged organelles under cellular stress conditions. Following sequestration in double ormultimembrane autophagic vesicles, the cargo is delivered to lysosomes for degradation. ATG5 is a keycomponentof an E3-likeATG12-ATG5-ATG16protein complex that catalyzes conjugationof theMAP1LC3protein to lipids, thus controlling autophagic vesicle formation and expansion. Accumulating data indicatethatATG5isaconvergencepointforautophagyregulation.Here,wedescribeascaffoldprotein,asanovelATG5interactorandanautophagyprotein.Usingseveralindependenttechniques,suchasYeastTwoHybridScreen, immunoprecipitation assays, immunoflourescence analysis, gel filtration tests and SILAC basedproteomicanalysis,weshowedthatthescaffoldproteininteractedwithATG5,andbothproteinsco-localizedinstress-responsivedot-likestructuresinthecytosol.Importantly,classicalautophagyinducers(starvationormTORblockage)stimulatedtheinteractionbetweenATG5andthescaffoldprotein.Moreover,weidentifiedthe critical aminoacid regionsof the scaffoldprotein for the interactionusing sitedirectedmutagenesis aswell asmolecular dynamics simulations. Knockdownof the scaffoldproteinor preventionof its binding toATG5 using mutagenesis blocked autophagy activation. Therefore, the scaffold protein is a new ATG5-interactingproteinandanimportantandnovelcomponentoftheautophagypathways.

Acknowledgements: This study is supported by TÜBİTAK 1001 Grant: 107T153 and TÜBİTAK-BIDEB 2211Scholarship.

Keywords: Autophagy, lysosome, RACK1, ATG5, ATG12-5-16, signaling, mTOR, p70S6K,proteinproteininteraction


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016

PP-2

LACCASE-GAMACDENCAPSULATEDPCLNANOFIBERSFORBETTERBIOCATALYTICACTIVITY

MehmetFatihCanbolat²,HasanBasriSavaş¹,FatihGültekin¹

¹|SuleymanDemirelUniversity,MedicalFaculty,MedicalBiochemistryDepartment.Isparta.Turkey.

²|SuleymanDemirelUniversity,TextileEngineeringDepartment,FacultyofEngineering.Isparta.Turkey.Theenzymes,asbio-catalysts,haveopenedadoortouseeco-friendly,greenandsustainableprocessesinthefield of synthetic chemistry and thereby getting results by natural methods in the production have beenpossible.Sincetheenzymescameoutofthereactionsthattheycatalyzedunaffectedandaresoexpensivecompounds, in order to enable their reuses and avoid undesirable mixtures with the post-productionproducts, the processes called immobilization should be carried out. Our study examines the effects ofcyclodextrinuseonenzymaticactivityfollowingenzymeimmobilizationintonanofibers.Analysishavebeenconducted on enzyme stability, enzyme activity and reaction performing potentials of enzymes.Electrospinningisacommonandversatilemethodinnanofiberspinningfromthepolymersolutions,polymermixturesorblends that canproduce fibers fromnanometer level tomicron level.Nanofibersproducedbyelectrospinningareformedbythecreationofpolymerdropletswhichisfollowedbythewhipping,stretchingand thinning of visco-elastic liquid under high electrical voltage application while after electrical fieldsurpassesthesurfaceenergyofthedroplet.Cyclodextrins(CDs),whichofferfunctionalsolutionsbycreatingthe complex structures (inclusion complex), have unique properties, i.e. have nontoxic nature, improvesolubility,andreduceundesiredodorandsomeothers.Wellknownandbroadlyusedlaccaseenzymewhichbelongstophenoloxidasegroupwaschosenasareferencematerialforthisstudy.Analysisthathavebeencarried out have not exactly confirmed the laccase-γCD inclusion complex formations, but it has beenobservedthatdifferentstructuralformationshavebeencreatedbycomplexformationsofadifferentkindofinteraction.Then,bothFTIRdataandtheimagesofcomplexsamplesobtainedhaveconfirmedthiscondition.Subsequently, It has been gained insight on whether the structures produced by electrospinning createsnanofiber or not by SEM analysis. Uniform fiber formations were observed and it was realized that fiberdiametershavebeen further thinned incasesofencapsulation.Nanofibersystemsandcyclodextrinuseonenzyme activity were analyzed and it has been indicated that the both nanofibers and cyclodextrin use,seperately,hasbeenpositivelyaffectedontheenzymeactivityandincreasedthestability.Moreoveritwasshown that the immobilization of the enzymes treated in physical mixture of γCD and laccase and thecreationof inclusioncomplex followingby introductionof thosematerials intoPCLpolymernanofibershasoccurredasignificantincreaseinthevaluesofenzymeactivation.Keywords:Laccase,longstability.


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PP-3

DECELLULARIZATIONOFLIVERANDHISTOLOGICALEXAMINATIONOFOBTAINEDMATRIX

AyşeYiğit²,BarbarosYiğit²,FuatUslusoy³,İlkayArmağan⁴ , HasanBasriSavaş¹

¹|SuleymanDemirelUniversity,MedicalFaculty,MedicalBiochemistryDepartment.Isparta.Turkey.²|SuleymanDemirelUniversity,MedicalFaculty,MedicalGeneticDepartment.Isparta.Turkey.³|SuleymanDemirelUniversity,MedicalFaculty,PlasticSurgeryDepartment.Isparta.Turkey.

⁴|SuleymanDemirelUniversity,MedicalFaculty,HistologyDepartment.Isparta.Turkey.Abstract In tissue engineering, the importance of the natural or synthetic scaffold in a conversion andtransplantationofthree-dimensionalstructureoftheproducedcellsislarge(1).Theformationofafunctionalvascular network resulting in new tissue is required for a successful tissue repair (2). Therefore using ofdecellularizated tissue as carrier support has advantages both in terms of comprising natural matrix andvascular supplying. In our study, decellularization methods of liver tissue for forming artificial liver andhistologic examination of obtained matrix. The liver tissues were fixed in 10% neutral formalin and thenembeddedinparaffinblocks.Sections(2-4μmthickness)wereobtainedusingaslidingmicrotomefromtheprepared paraffin blocks. These sections were stained by Hematoxylin-Eosin (H-E). Later, the liver tissuesectionswereanalyzedinthephotomicroscope(Fig1,2,3).HistologicalevaluationwithH-Estainingrevealedlessnucleiorcytoplasmicstaining indecellularizegroups (Fig2,3)comparedtonormal rat liver (Fig1).Weshowedthat itwasespeciallyobservednoticeablereductionhepatositafterthe5-hourdesellülarization. Inthisstudy,itshownthatthematrixwithvascularnetworkwhichisveryimportantfortissue-engineeringcanbeobtainedintheliver,withaneasyway,bydesellülarizationoftissue. Inthisway,producedhepatocytescanbeeasilyclingingonanaturalmatrixandproliferating(3,4).Figures:Fig1.Control-liver,Normalliverhistology(H-E,x10).

Fig2. 2-hourdecellularization-liver,moderateparenchymalstaining(H-E,x10).

Fig3. 5-hourdecellularization-liver,lessparenchymalstaining(H-E,x10).

Keywords:Keywords:Decellularizatedliver,tissueengineering.


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PP-4

ASSOCIATIONTRANSIENTRECEPTORPOTENTIALMELASTATIN2GENEPOLYMORPHISMSWITHPRETERMBIRTH

BelginAlaşehirli1,ReyhanGündüz2,ŞenizDemiryürek3,SerdarÖztuzcu4,ElifOğuz5,MeteGürolUğur2,AbdullahT.Demiryürek1

¹|DepartmentofMedicalPharmacology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

²|DepartmentofObstetricsandGynecology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

³|DepartmentofPhysiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey4|DepartmentofMedicalBiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey5|DepartmentofMedicalPharmacology,FacultyofMedicine,HarranUniversity,63300Sanliurfa,Turkey

Objective: Preterm or premature birth is defined as delivery of an infant before 37 completed weeks ofgestation. Preterm birth is the leading cause of neonatal death and infant mortality, often as a result ofrespiratory distress syndrome due to immature lung development. Transient receptor potential (TRP)channelsarenon-selectivechannelspermeabletomonovalentanddivalentcations.TRPmelastatin(TRPM)2channel canbe activatedbymicromolar levels ofH2O2andagents that produce reactiveoxygen/nitrogenspecies,providingadirectlinktoinflammation,oxidativestress,andcelldeath.Theaimofthisstudywastoinvestigate a possible association between TRPM2 gene polymorphisms and preterm birth in a Turkishpopulation.MaterialandMethod:Atotalof90womeninpretermlaborand94womenintermlaborwithsimilarageandsex were enrolled to this study. Genomic DNA from the participants was analyzed by a BioMark 96.96dynamic array system (Fluidigm, South San Francisco, CA, USA). For calculation of the significance ofdifferencesingenotypeandallelefrequencies,thechi-squaretestorFisher’sexacttestwasused.Results:Weobserved that theCCgenotype (15.3%vs.1.2%,p=0.0034)andCallele frequencies (28.8%vs.19.0%, p=0.048) of TRPM2 rs1612472polymorphismwerehigh in pretermbirth groupwhen compared tocontrols. There were significant changes in the genotype (TT, 60.9%; TC, 25.3%; CC, 13.8%) and allele (T,73.6%;C,26.4%)frequenciesfortheTRPM2rs933151polymorphisminpretermbirthwhencomparedtothecontrols(TT,56.4%;TC,11.7%;CC,31.9%,p=0.0038;T,62.2%;A,37.8%,p=0.0285).However,noassociationwasfoundwiththeTRPM2rs1618355polymorphism.Conclusion:OurresultsarethefirsttodemonstratethatTRPM2genepolymorphismsmaymodifyindividualsusceptibilitytopretermbirthintheTurkishpopulation Keywords:polymorphism,pretermbirth,TRPM2


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PP-5

ASSOCIATIONTRANSIENTRECEPTORPOTENTIALMELASTATIN2GENEPOLYMORPHISMWITHPREECLAMPSIA

BelginAlaşehirli1,ZekiyeDoğantürk1,ElifOğuz2,SerdarÖztuzcu3,ŞenizDemiryürek4,Reyhan

Gündüz5,MeteGürolUğur5,AbdullahT.Demiryürek1

¹|DepartmentofMedicalPharmacology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,

Turkey²|DepartmentofMedicalPharmacology,FacultyofMedicine,HarranUniversity,63300Sanliurfa,Turkey³|DepartmentofMedicalBiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

4|DepartmentofPhysiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey5|DepartmentofObstetricsandGynecology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,

Turkey Objective:Preeclampsiaischaracterizedbymaternalhypertension,proteinuria,oedemaand,in30%ofcases,byintrauterinegrowthretardation.Theprecisefactorsinvolvedinthepathogenesisofpreeclampsiaremainunclear and it is considered as a multisystem disorder. The oxidative stress, resulting from deficientremodelling of spiral arteries, is an important consequence of preeclampsia. The Ca2+ homeostasis isperturbed in preeclamptic placentas, most likely caused by a high oxidative stress level and lack of ATP.Transientreceptorpotential(TRP)channelisanonvoltage-gatedCa2+-permeablecationchannelsuperfamilyactivated by a variety of physicochemical stimuli. TRP melastatin (TRPM) 2 channel has been found torespondtooxidativestressbyincreasedchannelactivity.TheaimofthisstudywastoinvestigateapossibleassociationbetweenTRPM2genepolymorphismsandpreeclampsiainaTurkishpopulation.MaterialandMethod:Atotalof94patientswithpreeclampsiaand94healthycontrolswithsimilarageandsex were enrolled to this study. Genomic DNA from the participants was analyzed by a BioMark 96.96dynamic array system (Fluidigm, South San Francisco, CA, USA). For calculation of the significance ofdifferencesingenotypeandallelefrequencies,thechi-squaretestorFisher’sexacttestwasused.Results:Thereweremarkedchangesinthegenotype(TT,67.0%;TC,26.6%;CC,6.4%)andallele(T,80.3%;C,19.7%)frequenciesfortheTRPM2geners933151polymorphisminpatientswhencomparedtothecontrols(TT,56.4%;TC,11.7%;CC,31.9%,p<0.0001;T,62.2%;A,37.8%,p=0.0002).However,noassociationswerefoundwiththeTRPM2rs1612472andrs1618355polymorphisms.Conclusion:Tothebestofourknowledge,theseresultsarethefirsttodemonstratethecontributionTRPM2gene variants in preeclampsia. Our data showed that TRPM2 gene rs933151 polymorphism may modifyindividualsusceptibilitytopreeclampsiaintheTurkishpopulation.Keywords:polymorphism,preeclampsia,TRPM2


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PP-6

CHARACTERIZATIONANDCYTOTOXICITYOFBORONNITRIDE(BN)NANOPARTICLES:EMPHASISONTOXICOGENOMICS

HasanTürkez¹,MehmetEnesArslan¹,ErdalSönmez²,MetinAçıkyıldız³,AbdulganiTatar²,Fatime

Geyikoğlu²

¹|ErzurumTechnicalUniversity²|AtatürkUniversity

³|Kilis7AralıkUniversityBoron nitride (BN) nanoparticles were synthesized chemically and characterized by using X-raycrystallography (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM) andenergy-dispersiveX-rayspectroscopy(EDX)techniques.BNnanoparticlesappliedonthehumanlungalveolarepithelial cell line (HPAEpiC). To evaluate cytotoxicity of BN nanoparticlesMTT, LDH and NR assays werecarriedoutafter72hoursincubation.AccordingtoMTTresultsIC20valueforW2BwasdeterminedtoisolatetotalRNAfromculturesandinvestigateinmicroarrayanalysis.Finally,microarraydatawereinvestigatedwithThe Database for Annotation, Visualization and Integrated Discovery (DAVID) analysis and functionalcategoriesforthesegenesrevealedtounderstandeffectsofBNonbiologicalpathways.Themainaimofthisarticleistorevealcharacteristics,livingcellinteractionsandanycytotoxiceffectofBN.In conclusion, therewere somanypublications about toxic effects of boron including compounds and themajority of these researches claim adverse outcome ofmolecules.Main aim of this project is to find outcharacteristicsandcytotoxicityofboronnitrite(BN)nanoparticles,andalsoovercomethegeneraljudgementaboutnotorietyofboronmolecules.Inaccordancewithallcellviabilitytests,MTT,LDHandNRanalysisgivesimilarresults,andallofthemconfirmthatappropriateamountofBNdoesn’thave lethalresponseontheHPAEpiCcells.Microarrayresultsputforthanticancer/apoptotic,anti-metastaticanddevelopmentregulatoryeffects of BN. Also, positive impact of the molecule on diabetes and stroke pathophysiology can beinferencedfromgeneanalysis.

"ThisresearchwassupportedbyNationalBoronResearchInstitute(BOREN)(Grantnumber:Ç0391)."

Keywords:BoronNitride(BN)Nanoparticles,Invitro,Geneexpression,Alveolarepithelialcells,Microarray


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PP-7

CYTOTOXICACTIVITIESANDCELLULARMECHANISMEXERTEDBYWALTERINESSIAMORGANICOBRAVENOMAGAINSTNEURONALCANCERCELLS

ÇiğdemÇelen¹,AyşeNalbantsoy¹,BayramGöçmen²

¹|DepartmentofBioengineering,FacultyofEngineering,EgeUniversity,Izmir,35100,Turkey

²|ZoologySection,DepartmentofBiology,FacultyofScience,EgeUniversity,Izmir,TurkeySnakevenomisacomplexmixtureofmanysubstances,includingtoxins,enzymes,growthfactors,activators,andinhibitors,withawidespectrumofbiologicalactivitiesanddesignedtoaffectvitalprocesses,suchasthefunctionofnervesandmuscles,theactionoftheheart,thecirculationoftheblood,andthepermeabilityofmembranes.Cancerandneurodegenerationareoftenthoughtofasdiseasemechanismsatoppositeendsofaspectrum;oneduetoenhancedresistancetocelldeathandtheotherduetoprematurecelldeath.Manyofthegenesassociatedwitheither cancerand/orneurodegenerationplaya central role in cell cycle control,DNArepair,andkinasesignaling.Cancertherapyisoneofthemainareasfortheuseofproteinpeptidesandenzymesoriginatingfromanimalsofdifferentspecies.Cytotoxiceffectsofsnakevenomhavepotentialtokilltumor cells. In this study,we investigated functional and activity of cobra crude venomofW.morgani onneuronalcancercells.Forthispurpose,wedeterminedcytotoxiceffectsofW.morganicrudevenomagainstU87MG,SHSY5Y,KELLYandSK-NAScellsbyMTTassay.Crudevenomshowedhighcytotoxiceffectonnervecells with IC50 values varying between 0,15 - 5,2 µg/ml according to the viability percent. This resultsindicated that W. morgani cobra venom have potential for further studies to determine mechanisms ofactionsIn conclusion, W. morgani venom contain a large number of pharmacologically highly active substancesthrougha specificmodeofactioneachwith thepotentialofbecomingapotentdrug.The formulationsorcombinationsof this venom for targeteddrugdelivery couldbeused to for cancerandneurodegenerativediseasestreatments.ThisstudyisongoingtodeterminemechanisticeffectsofW.morganivenomonCXCR4expressionandNa+/K+-ATPaseactivitybyflowcytometer. Keywords:Walterinnessiamorgani,SnakeVenom,Neurodegeneration,Neurotoxin


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PP-8

EXPRESSIONPROFILINGANDPATHWAYANALYSISOFIRONOXIDE(FE2O3)NANOPARTICLESTOXICITYONHUMANLUNGALVEOLAREPITHELIALCELLLINE(HPAEPIC)

USINGMICROARRAYANALYSIS

HasanTürkez¹,MehmetEnesArslan¹,ÖzlemÖzdemir¹,MetinAçıkyıldız²,ErdalSönmez³,AbdulganiTatar³

¹|ErzurumTechnicalUniversity

²|Kilis7AralıkUniversity³|AtatürkUniversity

The main aim of this work to find out toxic effects of Fe2O3 on gene expression patten and pathwayrelationshipsofhumanlungalveolarepithelialcells(HPAEpiC).Chemically synthetized Fe2O3 was characterized via using X-ray crystallography (XRD) and transmissionelectronmicroscope (TEM) techniques. Cell viability and cytotoxicity were determined by 3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT), neutral red (NR) and lactate dehydrogenase (LDH)releasetest.Wholegenomemicroarrayexpressionanalysiswasperformedtobeabletofindouttheeffectsof Fe2O3on gene expression inHPAEpiC cell cultures. For further analyses, these geneswere functionallyclassified by usingDAVID (TheDatabase for Annotation, Visualization and IntegratedDiscovery)with geneontology(GO)analysis.AccordingtocytotoxicityassaysLC20valueforFe2O3is20.451mg/LandthisvalueisenoughtocallFe2O3nanopartclehightoxicitymolecule.DAVIDannotationanalysisindicatedthatFe2O3mediatedtoxicitydirectlyor indirectly affects regulation of cell proliferation, response to hormone stimulus, estrogen stimulus,cytokineactivityandbloodcirculationbyactivatingdiversegenes.

"ThisresearchwassupportedbyNationalBoronResearchInstitute(BOREN)(Grantnumber:Ç0391)."

Keywords: Iron oxide (Fe2O3) nanoparticles, Microarray Analysis, Toxicogenomics, Human Lung AlveolarEpithelialCells(HPAEpiC)


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PP-9

CYTOTOXICANDANTI-ADHESIVEPROPERTIESOFPOLYLACTIC-CO-GLYCOLICACIDCOATEDPOLYPROPYLENEMESHES

BaşakAru¹'²,VildanSanko³,ÜmranAydemirSezer⁴,SerdarSezer³,GülderenYanıkkaya

Demirel²'5

¹|MolecularMedicineDepartment,InstituteofHealthSciences,YeditepeUniversity,Istanbul

²|ImmunologyDepartment,YeditepeUniversitySchoolofMedicine,Istanbul³|InstituteofChemicalTechnology,TUBİTAKMarmaraResearchCentre,Kocaeli

⁴|MaterialsInstitute,TUBİTAKMarmaraResearchCentre,Kocaeli5|StemCellLaboratory,YeditepeUniversityHospital,Istanbul

Object: Abdominal adhesions are frequently observed complications such as trauma, infection, pain andfunction disorder after intraperitoneal and pelvic operations. These complications aremajor problems forpatients during healing process after such operations.Therefore anti-adhesive membranes are employedduring operations in order to prevent or diminish dangerous complications. Synthetic meshes wereintroducedandusedmore than fortyyears.Polypropylene (PP)meshcouldbeusefulas implantablemeshstructures in surgery due to its high durability and elasticity. On the other hand, it can lead post-surgicaladhesionafterimplantation.Bilayermeshesareusedinclinicalapplicationstopreventadhesion.MaterialandMethod:Inthisstudy,bilayermeshescomposedofelectrospun(polylactic-co-glycolicacid(plga)andplga-chitosan composite) coatingon thePPmeshweredeveloped.After incubatinghumanperipheralmononuclearbloodcellswithmeshesfor72hours;annexinV–propidiumiodidetestforevaluatingcytotoxicpotential ofmesheswas used. Formeasuring anti-adhesive properties ofmeshes, human fibroblastswereseededonmeshcoatedtissuecultureplatewellsandafter72hoursofincubation,LDHtestwasperformed.Testswereperformedastriplicates.Results:After72hoursofincubation,decreasedratesofnecroticcellsforbothbilayermesheswereobservedcomparedtoPPmesh,whereasapoptoticcellrateswerehigherforbilayermeshes.Comparedtononcoatedtissueculturewells,decreasedratesofadheredfibroblastswereobservedforallmeshes.Conclusion:PPmeshisanano-sizedproducthashighsurfaceareaporosityanditisanexcellentreplacementforanaturalextracellularmatrix.Thisdesignwouldpreventpost-surgicaladhesioninadditiontoincreasingtissue-materialintegrity. Keywords:Polypropylene,Polylactic-co-glycolicAcid,Adhesion,Cytotoxicity


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PP-10

CASPASE-8AND-9MEDIATEDAPOPTOSISINMUCOSALDISEASE(MD)FATALVARIANTOFBOVINEVIRALDIARRHEA(BVD)

B.TaylanKoç¹'²,NihatToplu³,E.TuğrulEpikmen³,T.ÇiğdemOğuzoğlu²

¹|AdnanMenderesUniversity,FacultyofVeterinaryMedicine,DepartmentofVirology

²|AnkaraUniversity,FacultyofVeterinaryMedicine,DepartmentofVirology³|AdnanMenderesUniversity,FacultyofVeterinaryMedicine,DepartmentofPathology

Object:Apoptosishavebeendefinedasprogrammablecelldeathinmanyliteratures.Generallyapoptosiscanbe induced by two major regulatory pathways. One of them is extrinsic apoptotic pathway by areceptor/ligand interaction mechanism which involves the recruitment of regulatory caspase-8 to deathreceptor complex. Intrinsic pathway is associated with caspase-9 and the further cleavage of effectorcaspases. Particularly, cancer and many diseases evade cell death mechanism and caspases. Bovine ViralDiarrheaVirus(BVDV)isauniqueagentforhealthofruminantsandpigsthatcauseshigheconomiclossesinbothlivestockanddairyindustry.BVDVhastwobiotypes,oneofthemisnon-cytopathogenic(ncp)blockstoprocess apoptosismechanism and no apparently induces cytopathogenic effects on cell culture. Thus, ncpbiotype causes persistent infection (PI) and infecting animals spreads to healthy animals throughout theirlives. Cytopathogenic (cp) biotype led to acute infection or superinfection in persistent animals named asMucosal Disease (MD), is highly fatal variant of BVDV. It is aimed to examine effects of both biotypesbelonging to MD, have been detected by Polymerase Chain Reaction (PCR) in blood sample of infectedanimal,oncellculture.Material andMethod:Molecular test andvirus isolationwereexamined fromblood sampleof an infectedcalf.Todetecttheexpressionsofcaspase-8and9by4’,6-diamino-2-phenylindole(DAPI)staininginMDBKcellculture.Cellculturefixationprocessareperformedat0.,12.,24.,48.,72.hraftervirusinoculationtodetectexpressionsofcaspase-8and-9 indifferenttimeperiod.Results: Inmicroscopicassessment,caspase-8andcaspase-9werestainedfrom12.hrandtheirexpressionsgraduallyincreasedinparalleltotime.Furthermore,thesefindingsshowedparallelismwithmolecularresults.Conclusion:Inconclusion,obtainedresults indicatedthatbothcaspase-8andcaspase-9pathwayscouldbeactivatedbyMD.ItistemptingtospeculatethatcaspasesactivitiesmaybesimilarinbothMDandacuteBVDinfections. Keywords:Keywords:Caspase-8,Caspase-9,Apoptosis,BVDV/MD,DAPI


4-7

May2

016

PP-11

EFFECTSOFARGANOILONMICRONUCLEUSANDMEGAKARYOCYTICEMPERIPOLESISINRATSEXPOSEDTOACRYLAMIDE

ZülalAtlıŞekeroğlu¹,VedatŞekeroğlu¹,BirsenAydınKılıç²

¹|OrduUniversity

²|AmasyaUniversityObject: Our study aimed to determine the effects of argan oil (AO) on the frequency of megakaryocyticemperipolesis(ME)andmicronucleus(MN)againstacrylamide(AA)-inducedtoxicityinrats.MaterialandMethod:TwentyhealthyratswereobtainedfromtheExperimentalAnimalCenterofUniversityofOndokuzMayıs(Samsun,Turkey).ThestudywasapprovedbytheMedicalResearchEthicsCommitteeofthatuniversity(HADYEK2014/24).Animalsincontrolgroupwereorallygavagedwithaconstantvolumeof1ml/kgbwof0.9%NaClsolution.WhileAAwasadministeredintraperitoneallyinadoseof50mg/kg/day,AOwasadministeredbyoralgavage inadoseof6mg/kg/dayeveryotherday for30days.AnimalswerealsotreatedwithmixtureofAA(50mg/kg/day)andAO(6mg/kg/day)everyotherdayfor30daysuntiltheywereeuthanized.BonemarrowsampleswereanalyzedforthefrequenciesofMEanMN.Results: AA significantly increased the formation ofME andMNanddecreased the ratio of polychromaticerythrocytes(PCEs)inbonemarrow.NosignificantdifferenceswereobservedintheanimalsreceivedtheAOcompared to the control group. Co-treatmentwithAA+AO significantly ameliorated theMN,MEandPCEsvaluesinbonemarrow.Conclusion:ThesefindingssuggestthatAOmayplayabeneficialroleinreducingthecelldamageinducedbyAA.Keywords:Acrylamide,arganoil,micronucleus,megakaryocyticemperipolesis,bonemarrow.


4-7

May2

016

PP-12

ND6INMITOCHONDRIALDNAMIGHTHAVEROLEONBETA-CELLDEATHTARGETINGBYMIR-29A

ZeynepÖztürk¹

¹|BingölÜniversity,FacultyofArtandScience,DepartmentofMolecularBiologyandGenetics,12000,Bingöl,

TURKEY DiabetesMellitus is a commonmetabolic disorder in theWorld. This disorder characterizedbyhighbloodglucoselevelandrelatedwithleakinsulinproductionbythepancreaticbetacells.Alsothebeta-celldeathisimportant forT2DM.Moreover, geneticplayers seem tobe involved in thedevelopmentofT2DMsuchasMiRNAs.MiRNAsarepost-transcriptionalregulatorsofproteinexpressionandtheybindtocomplementarysequences in the 3’UTRs of their target mRNA either performing transcript degradation or translationalinhibition.Itisacceptedthattheyplayaroleinthedefectofβ-cellstosecreteenoughinsulinsotriggertype2diabetesmellitus(T2DM).ThisprojectfocusesonmiR-29athathasbeenshowntobeinvolvedinglucose-inducedbeta-celldysfunction,previously.RecentlytwopublicationshaveshownthatmicroRNAsarepresentinthemitochondria,suggestingthatmicroRNAscanregulatemitochondrialgene-expression.Sointhisstudythe target sequence was searched for miR29-a for mitochondrial gene ND6. ND6 is a vital gene formitochondrialfunctionandifmitochondrialfunctionisdamagedinbeta-cells,betacelldysfunctionanddeathare observed. Asmethod, the potential targets ofmiR-29a inND6 sequencewas identified and the oligosweredesigned. Then theseoligoswere transferred into a vectorwhichhas luc2 gene. These vectorswerecopiedinE.colicellsthentransfectedtoHEKcellsforluciferaseassay.Theprimaryaimoftheluciferaseassaywastodetermine,whethermiR-29ainteractswiththepredictedtargetsiteinND6.Resultsoftheluciferaseassay’sdataandt-testshowedthattherewasnotahighqualitycomplementationinND6betweenmiR-29a.Butforsayacertainanswerthisexperimentmustrepeatwithanothercandidatetargetsites.Forthefuturethe existing connection to T2DMbetweenmiR-29a andND6 because ofmitochondrial functions could bemotivating.Keywords:miR-29a,microRNA,mtDNA,diabetesmellitus,Beta-celldeath.


4-7

May2

016

PP-13

EFFECTOFOZONETREATMENTONRADIATIONCOLITISINRATS

AyhanKutlu¹,EsraErdoğan¹,BülentUysal²,RecepGümüş¹,EsinGündem³,ÖmerSager³,MuratBeyzadeoğlu³,EminÖztaş¹

¹|DepartmentofMedicalHistologyandEmbryology,GulhaneMilitaryMedicalAcademy,Ankara,Turkey

²|DepartmentofPhysiology,GulhaneMilitaryMedicalAcademy,Ankara,Turkey²|DepartmentofRadiationOncology,GulhaneMilitaryMedicalAcademy,Ankara,Turkey

Object:Radiationcolitisoccurringafterradiationtreatmentofcancerandamajorsideeffectsthatreducethepatient'squalityoflife.Theformationofradiationcolitisisthoughttobecausedbyoxidativemechanism.Weaimedtoinvestigatetheefficiencyofintraperitonealozoneapplicationofradiation-inducedcolitisinrats.Material andmethod: 42Wistar albino ratswere divided into 5 groups including Sham, Radiation,Ozone,Radiation+OxygenandRadiation+Ozone.Shamgrouphadnoapplication.MedicalozonewasadministeredtoOzone group intraperitoneally 1 mg/kg/day dose the third day to the seventh day. 25 Gray gamma rayexposurewasapplied to theabdominal-pelvic regionofRadiation,Radiation+OxygenandRadiation+Ozonegroups. All groupsmade no application for two days. Exposure of radiation from day 3th to day 7th at 1mg/kg/daydoseofoxygenwasadministrated toRadiation+Oxygengroupandwasadministratedozone toRadiation+Ozonegroup.TherehasbeennoapplicationuntiltheendoftheexperimenttheRadiationgroup.TUNEL staining was performed to determine apoptosis occurring in the intestinal tissue. The differencebetweengroupswasevaluatedsemi-quantitatively.Percentageofpositivecellstoall thecells inthe imageareawasevaluatedunderalightmicroscope;-(None),+1(little),+2(medium),+3(severe)asscored.Results: The immunohistochemical evaluation; apoptotic cells was not observed in the groups withoutradiation. Apoptotic cells score were severe(+3) in Radiation and Radiation+Oxygen groups, little(+1) inRadiation+Ozonegroup.PercentageofapoptoticcellsreductionintheRadiation+OzonegroupaccordingtotheradiationandRadiation+Oxygengroupswasstatisticallysignificant(p<0,05).Conclusion:Wedemonstratedthatmedicalozoneapplicationreducedseverityofradiation-inducedcolitisintheratintestinalepithelial.Wefurtherdemonstratedthatmedicalozonepossessedbothanti-apoptoticandanti-inflammatory properties after radiation injury. These findings suggested the potential role ofmedicalozoneagainstintestinalepitelialinjuryduringradiotherapy. Keywords:Keywords:Apoptosis,Ozone,RadiationColitis,Radiotherapy


4-7

May2

016

PP-14

EFFECTSOFCURCUMINANDQUERCETINONREACTIVEOXYGENSPECIESANDAPOPTOTICPROCESSINCHRONICMYELOIDLEUKEMIA(K562)CELLS

ErgülMutluAltundağ¹'²,AyşeMineYılmaz¹'²,SemraKoçtürk²'³,YavuzTaga¹'²,A.SühaYalçın¹'²

¹|DepartmentofBiochemistry,SchoolofMedicine,MarmaraUniversity,Maltepe,34854Istanbul,Turkey.²|GeneticandMetabolicDiseaseResearchCenter,MarmaraUniversity,Maltepe,34854Istanbul,Turkey.³|DepartmentofBiochemistry,SchoolofMedicine,DokuzEylülUniversity,Inciralti,35340Izmir,Turkey.

Aim:Wehaveusedquercetinandcurcumin,twonaturalpolyphenols,toinduceapoptosisofchronicmyeloidleukemiacells.Curcuminhaslimitedclinicalusebecauseofitslowbioavailability(<2%).Therefore,wehavetriedsynergisticcombinationofcurcuminwithquercetintoincreaseitsapoptoticeffects.Methods: Cell proliferationwas analyzed byWST-1method and IC50 valueswere determined. Synergisticeffects of the two polyphenols were analyzed by the CalcuSyn combination analysis program. Annexin-Vstainingwasusedfordetectionofapoptosis,DCFDAwasusedfordetectionofreactiveoxygenspeciesandJC-1 dye was used for detection of mitochondrial membrane potential. All of the above analyses wereperformedbyflowcytometry.Fluorometricassaywasusedtomeasureintracellularglutathione.ChromatincondensationwasshownusingHoechst33342dyebyflorescencemicroscope.ProteinexpressionsassociatedwithapoptoticmechanismswereanalyzedusingtheWesternblotmethod.Results:Therateofapoptosiswasenhancedbythesynergisticcombinationofquercetinandcurcumin.Usingthecombinationdoses thepotencyofquercetinandcurcuminwasdecreased1.6 to7.3 fold, respectively.Quercetin and curcumin caused concentration-dependent decrease in cell proliferation and inducedapoptosis (%92.48) inCMLcellsafter48hours.Combinationofquercetinwithcurcuminreducedbothcellviabilityand intracellularGSH (%17.2).However, increasedROSandmitochondrialmembranepotentialaswell as apoptosis rate was noted. The analysis of variance (ANOVA) was used for the comparison of allexperiments.Conclusion: We suggest combined use of the two polyphenols, quercetin and curcumin to increase theirbioavalibility. Our results indicate that quercetin and curcumin exhibit a high level of strong synergism incombination,withenhancedbioactivitytherebyreducingtherequiredeffectivedoseforeachagentKeywords:Chronicmyeloidleukemia(CML),quercetin,curcumin,apoptosis,reactiveoxygenspecies(ROS).


4-7

May2

016

PP-15

CANTAURINEPREVENTIFOSFAMIDENEUROTOXICITY?

ÖzgeBulut²,HafizeSedaVatansever¹'³,FatmaTaneli⁴,YeşimGüvenç⁴,RemziyeKendirci¹,RaziyeYıldız⁴,AykanÖzgüven²

¹|DepartmentofHistologyandEmbryology,FacultyofMedicine,CelalBayarUniversity,Manisa,Turkey

²|CelalBayarÜniversity,FacultyofMedicine,DepartmentofPediatricHematologyandOncology,Manisa,Turkey

³|NearEastUniversity,ExperimentalHealthScienceResearchCenter,Nicosia,NorthCyprus4|CelalBayarÜniversity,FacultyofMedicine,DepartmentofBiochemistry,Manisa,Turkey

Ifosfamide is often used in the treatment of childhood cancer. Ifosfamide have central nervous systemtoxicity such as changes in consciousness, cerebral infarction due to seizures, paralysis, neuropathy,leukoencephalopathyandototoxicity. Inthisstudy,weaimedtohistologicalanalysesofTaurinebecauseofantioxidantpropertiesontheeffectof ifosfamideneurotoxicityandnephrotoxicity.Wistar-Albinoratsweredivided into 4 groups;Group 1was given intraperitoneal injection of 50mg / kg ifosfamide,Group 2wasgiven intraperitoneal injectionof50mg / kg ifosfamide+1g / kgoral taurine (7days),Group3wasgivenintraperitoneal saline (control group), Group 4 was given oral 1 g / kg taurine (7 days). All animals weresacrificed after 8 days of studies and brain tissue were fixed in 10% formalin solution. After paraffinembedding procedure, sections were stained either TUNEL for apoptotic cell detection, or indirectimmunoprexidase staining fordistributionsofBax, cytochrome-C, caspase-3, caspase-8, i-nos,e-nosandn-nos. After histochemical analyses, in ifosfamide given group, chromatin condensation of neurons andneurogliacells,edemaaroundtheneurogliacellsanddemyelinationofoligodendrocytecellswereobserved.In IfosfamideandTaurinetreatedgroup,chromatincondensationofneuronsandneurogliacellswererare.The number of TUNEL positive cells was higher in ifosfamide given group when compared with other.Increased caspase-3 and i-nos immunoreactivity were also detected in ifosfamide given group.While baximmunoreactivitywasnegative inneurons in ifosfamidegivengroup,onlyweakBax immunoreactiivtywasobserved inneurogliacells.CytochromeC,caspase-8,e-nosandn-nos immunoreactivitieswerenegative inall groups. In conclusion; ifosfamide induced apoptotic pathways expressiong of Bax, caspase-3 and i-nos.Taurinemayprotectsifosfamideneurotoxicityinhibitingcellulardamage. Keywords:apoptose,caspase-3,NOS


4-7

May2

016

PP-16

THEEFFECTSOFZIZIPHUSJUJUBAONSKINCANCERCELLS

VesileDüzgüner¹,AltuğKüçükgül²,M.Mustafaİşgör²,MustafaCellat²,PınarKızılkaya¹

¹|ArdahanUniversity²|MustafaKemalUniversity

Objects:Reactiveoxidantspeciesmayresultinoxidativestressinthecellularandextracellularenvironmentand have been implicated in the etiology and progression of many diseases mainly chronic. Cutaneousmelanoma is one of the most serious skin cancers. It is caused by neural crest-derived melanocytes -pigmentedcellsnormallypresentednormallyintheepidermisand,sometimes,inthedermis.ZiziphusjujubaMill. (ZJ) distribute in the tropical and subtropical regions of Asia and have been employed as essentialoriental folkmedicine for thousandsof years. This studywas carriedout to investigate theanticancerandantioxidanteffectsofZJonmelanomacells.Materials Methods: Cell survival was quantified by colorimetric MTT assay with time and dose response.Melanomacellsweretreatedwith100µmolZiziphusjujubaessentialoilforthreehours.Themorphologyofcells were monitored and pictured. Then, the cell homogenates were taken after treatment period.Glutathione (GSH), Total oxidant capacity and total antioxidant capacity (TOC, TAC) and nitric oxide levelswereestimatedusinghighlyspecificspectrophotometricmethods.Results: Ziziphus jujuba inhibited growth and proliferation, and increased total antioxidant capacity. MTTassays indicated that Ziziphus jujuba significantly decreased cell viability. The results demonstrated thatZiziphusjujubapreventeddecreaseinantioxidant levelsandnitricoxidelevels inmelanomacells.AlsoGSHlevelswereimprovedafterZJtreatment.Conclusion: Ziziphus jujuba demonstrated potent antiproliferative and antioxidative effects in melanomacells.ThestudyprovidesascientificandethnopharmacologicalrationaleforthetherapeuticuseofZJfruitKeywords:Keywords:Ziziphusjujuba,Melanoma,Oxidativestress


4-7

May2

016

PP-17

INVITROEFFECTSOFOLEUROPEINONMELONOMACELLS

AltuğKüçükgül¹,M.Mustafaİşgör¹,VesileDüzgüner²

¹|MustafaKemalUniversity²|ArdahanUniversity

Objects:Oxidativestressisconsideredtobeinvolvedinthepathophysiologyofallcancers.Melanomaisthemaincauseofdeathinpatientswithskincancer.Itiscausedbyneuralcrest-derivedmelanocytes-pigmentedcells normally presentednormally in theepidermis and, sometimes, in thedermis.Oleuropein is themainphenoliccompoundofolivetreeandisresponsibleforthecharacteristicbitternessofolivefruits.Oleuropeinisaheterosidicesterofelenolicacidandhydroxytyrosolandpossessesbeneficialeffectsonhumanhealth.Theaimofthisstudywastodetermineinvitroeffectsofoleuropeinonmelanomacells.MaterialsMethods:ViabilityofthecellswasquantifiedbyMTTassayinatimeanddoseresponsemanner.Melanomacellsweretreatedwith100µmolforthreehours.Themorphologyofcellswasmonitored.Then,thecellhomogenatesweretakenaftertreatmentperiod.Glutathione(GSH),Totaloxidantcapacityandtotalantioxidantcapacity(TOC,TAC)andnitricoxidelevelswereidentifiedusingspecificcolorimetricmethods.Results:Oleuropeintreatmentincreasedcellviabilityinmelanomacelllineinadose-dependentmanner.Thecellstreatedwitholeuropeinwithoptimumdoseandtime.OleuropeininhibitstheactivationofnitricoxideandpreventeddecreasedlevelsofGSH.Alsototaloxidantcapacitydecreasedsignificantlyaftertreatment.Conclusion: Oleuropein decreased cell death and triggerred antioxidant system positively. These findingssuggested that oleuropein has potent anticancer and antioxidant properties onmelanoma cells. However,furtherinvivostudiesarerequiredtodeterminetheexactpotentialofthisagent.Keywords:Keywords:Oleuropein,Melanoma,Oxidativestress


4-7

May2

016

PP-18

PROTECTIVEEFFECTSOFPLANTAGOHOLOSTEUMSCOP.EXTRACTONHYDROGENPEROXIDE-INDUCEDDAMAGEINL929FIBROBLASTINRELATIONTOTHEANTIOXIDANT

ACTIVITY

YasinGenç¹,Ü.ŞebnemHarput¹

¹|HacettepeUniversity ThegenusPlantago(Plantaginaceae)isrepresentedby21speciesinTurkey.Severaleffectsaredescribedforthe genus Plantago such as antitumoral, anti-inflammatory, antifungal, antibacterial, analgesic,antispasmodic,antiviralandhepatoprotective.Plantagospeciesareknownnotonlyasafoodplant,butalsoanoldmedicinalplantthathasbeenusedexternallytotreatmentofwound,abscessandacnes,internallytotreatment of diabetes, urinary infections and cancer as a decoction, common cold and viral infections asinfusion inAnatolia.Earlier investigationsperformedonPlantagospeciesresulted inthe isolationofmainlyiridoid glucosides, phenylethanoid and flavonoid glycosides, caffeic acid derivatives, polysaccharides andlipids.Woundhealingconsistsofdifferentstagesofinflammation,proliferationandremodelingstage.Inthisstudy; antioxidant, proliferative and protective effect of extract were investigated against H2O2 damagedL929murine fibroblasts to understand wound healing properties of the extract. As a result of our study,water extract of P. holosteum showed radical scavenging activity against DPPH,NO, SO andABTS radicalscomparabletothatofknownantioxidantsBHAandascorbicacidandwaterextractofP.holosteumdidnotincrease the proliferation of the fibroblast in the concentration range of 10-200 mg/mL. In the case ofprotective effect of the extract against H2O2 injury, cytotoxicity of hydrogen peroxide was found dose-dependent in L929 fibroblasts.While pre-incubationwith the extract protects fibroblasts against oxidativedamageofH2O2,incubationwiththeextractafterH2O2applicationdidnotrepairtheoxidativeinjury.Thisresult correlateswith the antioxidant potential of the extract.Our study on P. holosteumwill continue toinvestigateotherparametersofwoundhealingindifferentcelllines.*U.SebnemHarputhasbeensupportedTUBA-GEBIP/2013awardKeywords:Plantagoholosteum,hydrogenperoxide,woundhealing,antiinflammatory


4-7

May2

016

PP-19

THENEUROTOXIC,CYTOTOXIC,APOPTOTICANDANTIPROLIFERATIVEACTIVITIESOFTHEEXTRACTSOFSOMEMARINEALGAEONNA2BCELLLINE

OğuzKurt²,FeyzanÖzdalKurt²,CelalMertAkçora²,MahmudÖzkut¹ ,İbrahimTuğlu¹

¹|CelalBayarUniversity,MedicalFacultyDepartmentofHistology-Embryology²|CelalBayarUniversity,FacultyofSciences&LettersDepartmentofBiology

Marine algae are natural compounds and their cytotoxic, antiproliferative and apoptotic effect has beenshownformanycancertypes.Anticancereffectofthesealgaemayalsoimportantnervoussystemtumours.Therefore,theseeffectsofthealgaeextractsonmouseneuroblastomacellline(NA2B)wereinvestigatedinculture.ExtractsfromPetaloniafascia,JanialongifurcaandHalimedatunawereharvestedintheAegeanSeashoresofTurkey.15000cell/ml/wellweretreatedbyalgaeextractsat1to0,00007µg/mldilutionratesforNA2B cell survival and proliferation by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT).Cytotoxiceffectof algaeextractson theNA2Bcellswerealso investigated foroxidative stresswithNitricOxideSynthase(NOS)immunocytochemistryandapoptosiswithterminaldeoxynucleotidyltransferasedUTPnickendlabeling(TUNEL).Moderatetoxiceffectwasexaminedbyneuriteinhibitionwithneurotoxicityscreeningtest(NST)attheIC50dilutionsofextracts.ItwasobservedbyMTTresultsthatJ.longifurcaextractsweremore toxic compared to P. fascia and H. tuna extracts. Therewas clear increase of endothelial andinducibleNOSimmunostainingforoxidativestressandTUNELforapoptosisafterextractsapplication.Therewas inhibition of neurite outgrowthwith a statisticalmeaning due tomoderate toxic effect of algae. Thisstudysuggestedthatthecompoundsofalgaecouldbeusefulfortheanticancertreatmentofnervoussystemtumour.Meanwhile,theirspreadonthemarineenvironmentmaydangerousforaquaticanimalsandalsoforhumanhealthbecauseofthefoodchain. Keywords:Cytotoxicity,Marinealgae,NA2Bcellline,Neuriteinhibition,Neurotoxicityscreeningtest,TUNEL


4-7

May2

016

PP-20

SERUMHYPOXIAINDUCIBLEFACTOR-1ALPHALEVELSINPATIENTSWITHPOLYCYSTICOVARYSYNDROME

ElifPolat¹,YaşarNuriŞahin¹,EsraÇınarTanrıverdi²,FatmaBetülÖzgeriş¹

¹|DepartmentofBiochemistry,AtaturkUniversityMedicalSchool

²|DepartmentofGynecologyandObstetrics,NenehatunMaternityHospital,ThePolycysticOvarySyndrome(PCOS),unknownetyology, ismajorhealthyproblem inwomenworldwide.Resently some studys have shown that hypoxia inducible factor (HIF)-1alpha, the oxygen-sensitivetranscriptionalactivator,playsakeyrole in thedevelopmentofmammalianovarian folliculardevelopmentandovulation.Inthisstudy,weaimedtoinvestigatetothelevelsofHIF-1alphainindividualwithPCOS.Thestudywascarriedouton48womenconsistedof28patientswithPCOSand20healthyonesascontrol. Inserumsobtainedfrombloodsamplestakenfrompatientandcontrolgroups,theHIF-1alphaconcentrationsweremeasuredbyELISAkit,aspecificenzyme-linkedimmunosorbentassay.Therewasnodifferentbetwenthe groups inmeanof agedistrubution, p>0.05. Itwas found that serumHIF-1 alpha concentrationswerehigherinpatientwithPCOSthanincontrolgroup(50.92±34.73,41.16±40.98,respectively)buttherewasnostatistically significantdifference,p>0.05.Becauseof there isno statistically significantdifferencebetweentwogroupsinourstudy,wesuggestthatarestudiedwithmorethannumberofsamples.


4-7

May2

016

PP-21

THEEFFECTSOFNMDARECEPTORSUBUNITSANDCHOLESTEROLONAMYLOIDBETATOXICITYINSHSY-5YCELLS

PınarAkan¹'²,G.ÖzlemÇalan¹,D.AyçaErsen3,UğurBora²,SemraKoçtürk¹

¹|DokuzEylulUniversity,Medicalfaculty,MedicalBiochemistryDepartment²|DokuzEylulUniversity,HealthScienceInstitute,NeuroscienceDepartment

3|DokuzEylulUniversity,Medicalfaculty,PathologyDepartment

Objective: There is an increased sensitivity to N Methyl D aspartate (NMDA) receptor and irreversibleneuronal cell death inAlzheimer’sDisease (AD). Increaseof intracellular calcium levels,whichoccurs afterNMDA receptor induction, can lead to neuronal cell death. It has been suggested that amyloid beta (AB)peptidesattoxicconcentrationsmayincreasepregnenolonesulfate(PS)levelinthepresenceofcholesterol.It’sknownthatmicromolarlevelofPShasanagonisticeffectonNMDAreceptor,whichhasaheterogeneousstructure.FortheeffectofPSonNMDAreceptor,acombinationofreceptorsubunits(NR2A,NR2BandNR1)isrequired.However,theroleofNMDAsubunitsandtheeffectofPSonABtoxicityinthepresenceofhighcholesterol levels has not to be revealed yet. The aim of this study is to determine the effects of NMDAreceptorsubtypesonABtoxicityandtoexaminepossibleconnectionwiththechangeofPSlevel.MaterialandMethod:SH-SY5Ycellsweretreatedwith10µMAB1-42afterthepeptideswere incubatedat37ºCforthreedaysandcholesterol(1.2-9mM).ToexaminetheeffectsofNMDAreceptorinhibitors,thecellswere also treated with NVP-AAM077 (NR2A inhibitor, 12nm), ifenprodil (NR2B inhibitor, 0.5 µM) andMK801/memantin (total inhibitor, 5µM) for 24, 48 and 72 h. Changes in cellular cholesterol and PS levelsweredeterminedsimultaneouslyinadose-andtime-dependentmanner.Thecellviabilitywasalsoevaluated.Results and Conclusion: The treatment together with cholesterol (9mM) and AB peptides decreased cellviabilitynearly50%ofcontrolandsignificantlyincreasedPSlevels(p<0.05).Thetreatmentofifenprodilandmemantin significantly protected SHSY-5Y cells from the toxicity induced by the treatment together withcholesterolandABpeptide(p<0.05).NMDAreceptorNR2BsubunitmayplayakeyroleinneuronalcelldeathinducedbyABpeptides. The changes in PS levels and itsNMDA receptor activitymay affect neuronal cellsurvival.

Keywords:NMDAreceptor,amyloidbetatoxicity,neuronalcelldeath,cholesterol,pregnenolonesulfate,NR2receptorsubunit


4-7

May2

016

PP-22

NUCLEICACIDDAMAGEINWOMENWITHPOLYCISTICOVARYSYNDROME:8-HIDROKSI2’-DEOKSIGUANOZINLEVELS

ElifPolat¹,YaşarNuriŞahin¹,EsraÇınarTanrıverdi²,NezahatKurt¹

¹|DepartmentofBiochemistry,AtaturkUniversityMedicalSchool

²|DepartmentofGynecologyandObstetrics,NenehatunMaternityHospitalPolycystic ovary syndrome (PCOS) is themost common gynecological endocrine disorder in reproductive-agedwomen.TheetiologyofPCOSisnotfullyunderstood.Freeradicalscausesdamageordeathtothecells.Hydroxyl radical (OH-) reactswithDNApurinebasesandgenetatespurine radicals.OxidativeDNAdamageusesasameasureofthedamageindicatorandusuallymeasuresas8-Hydroxy-2ʹ-deoxyguanosine(8-OHDG)nucleosides.Inthisstudy,weaimedtoinvestigateforthelevelsof8-OHDGinpatientswithPCOS.Thelevelsof8-OHDG,FSH,LHwereassessedin103patientswithPCOSand47healtycontrolgroup.IsolationofDNAfrom all blood samples was performed used by manifacture kit. 8-OHDG levels were studied with highperformance liquid chromatography (HPLC-UV and -EC). Because variables showed abnormal distribution,Manny Whitney U test, nonparametric test, was applied for staistical analysis in the present study. Asstatistical, p< 0.05 values were considered significant. The levels of FSH and LH in PCOS group weresignificantlyhigher than control group,p<0.05. Therewas significantlydifferent in8-OHDG levelsbetweenPCOSandcontrolgroups(1.96±0.83,0.45±0.15respectively),p<0.001.Accordingtotheresults,ourstudywasshownthat8-OHDGlevelswereincreasedinpatientwithPCOS.Wesuggestthatlevelsof8-OHDGmaybeevaluetedasanindicatorofoxidativestressinPCOSKeywords:8-OHDG,PCOS


4-7

May2

016

PP-23

THEEFFECTSOFLIQUIDAMBARORIENTALISOILONHUMANGLIOSBLASTOMACELLSINH2O2INDUCEDOXIDATIVESTRESS

M.Mustafaİşgör¹,AltuğKüçükgül¹,VesileDüzgüner²,AydınAltop³,MeryemNurAtabay¹,

ZeynepYiğit¹,AzimeKüçükgül4

¹|MustafaKemalÜniversitesi²|ArdahanÜniversitesi

³|OndokuzMayısÜniversitesi4|TunceliÜniversitesi

Objects:Glioblastomas, themostmalignant form,arecharacterizedby increasedproliferationand invasion into thesurroundingnormalbraintissue.TheessentialoilofLiquidambarorientalisvar.orientalis,anendemictreespeciesinTurkey, has medicinal and cosmetic properties; and its antioxidant properties were investigated in H2O2 inducedoxidative stress in human glioblastoma cells. Materials Methods: Cell survival was quantified by colorimetric MTTassaywithtimeanddoseresponse.U87humangliblastomacellswerepretreatedwithH2O2100µMafter30minutes,100 nM Liquidambar orientalis essential oil was added to the cells for three hours. Themorphology of cells weremonitored and pictured. Then, the cell homogenates were taken after treatment period. Glutathione (GSH), Totaloxidantcapacity(TOC)andtotalantioxidantcapacity(TAC)andnitricoxidelevelswereestimatedusinghighlyspecificspectrophotometric methods. Results: We demonstrated that the treatment of glioma cells with LiquidambarorientalisantagonizedH2O2inducedoxidativestress. InU87cellsexposedtoH2O2,GSHwassignificantlydepleted.The results suggest that Liquidambarorientalis regulatednitric oxide andTOC levelswhichwas increasedbyH2O2induction.TACwasincreasedaftertreatmentofoil.Conclusion:Liquidambarorientalistriggersantioxidantsysteminhumangliomacells.Thepresentfindingsmayhaveusefulimplicationsforthepotentialuseoftheseoilinthemedicalfieldaswellasinthefoodindustry


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PP-24

EVALUATIONOFVITAMIND,VITAMINB12,IRONLEVELSINASMALLGROUPOFMEDICALFACULTYSTUDENTS

SalihaAksun¹,Alperenİhtiyar¹,HasanOrhanÇetin,GamzeKıvrak²,FidanBulut²,HakkıMert

Keklik²,CanHepduman²,BülentÖzkan³,RecepSütcü¹

¹|İzmirKatipCelebiUnivercityMedicalFaculty,DepartmentofMedicalBiochemistry²|İzmirKatipCelebiUnivercityMedicalFaculty,3rdClassStudents

³|İzmirKatipCelebiUnivercityMedicalFaculty,DepartmentofBıostatisticsAim:HumansgetvitaminDfromexposuretosunlightanffromdiet.InyoungpeoplevitaminDisimportantfor calciumabsorptionandbonegrowth. Low levelsof vitaminD is found tobe related to somediseases.Cancer,skeletaldiseases,diabetesmellitus;multiplesclerosisandalsoitisaboutmenthaldevelopment.WewonderedwhatisvitaminDlevelsinourmedicalfacultystudents,howoftendotheybenefitfromdaylight.Andwealsoevaluatedtheirsomeotherbiochemicalparameters.Methods: Initially 60medical faculty students joined to work. They were divided according to living withfamily,onceagaindividedgirlsandboys.Results:Inmalestudentslivingwiththeirfamily,VitaminDlevelshigherthanothers.B12,ıron,hematologicparametersfoundsimilar inallgroups.VitaminDcutoffvalueisknownas20ng/ml.Whendividedtoonlycuttoff, inhighvitaminDgroup(34,48ng/ml) , it isfoundthatB12(372,4pg/ml), ferritin(56,95ng/ml)andiron(88,70 µg/dl) average levelswere higher than low vitaminD groups (12,50 ng/ml) respectively 349,71pg/ml,49.85ng/ml,79,5µg/dl.İnhighvitaminDgroupthedurationofexposuretodirectsunlightis2,7hourwhen compared others 2.22 hours in day. we also measured Superoxid dismutase, glutatyon peroxidase,totalantioxidant(TAS)andtotaloxidant(TOS)capacity.Conclusion: It is important tobeawareofbenefitofvitamin forhealthy lifeandsoeveryoneshouldknowtheirownbiochemicalstate.Keywords:Keywords:Liquidambarorientalisvar.orientalis,U87-MG,Oxidativestress


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016

PP-25

ANTICANCEREFFECTOFSOMESILVER(I)-N-HETEROCYLICCARBENECOMPLEXESONDIFFERENTCANCERCELLLINES

IşılYıldırım1,AydınAktaş2,TürkanKutlu1,YetkinGök2

¹|InonuUniversity,DepartmentofChemistry/Biochemistry,44280,Malatya/Turkey

²|InonuUniversity,DepartmentofChemistry/Organicchemistry,44280,Malatya/Turkey

Metal-baseddrugshavemanyeffectiveinthetreatmentofavarietyofdiseasesespeciallyforcancer.So,themetallo-organicdiagnosticagents’attentionhasbeenincreaseinrecently.Althoughmetal-containingdrugswhichsuchascisplatin,fluorouracilhaveknownaseffectivetherapeutics,Agareoneofthemetallo-organiccompoundsthatseeinterestinlastfiveyears.ThisworkwastoinformofanticanceractivityofsomeAg(I)-N-Heterocycliccarbene(NHC)complexesagainstdifferentcancercells.

Liuandcoworker identified in theirwork thatsomeAg(I)-NHCcomplexesanticancereffects againstbreastcancers (MCF-7 and MDA-MB-231) and colon carcinoma (HT-29) cells. While, the 4-hydroxyl substitutedcompoundexhibited antiproliferative activitieswith IC50 9.2-16.2µM, the fluoro andmethoxy-substitutedcompoundswereanticanceractivitiesinthethreecancercelllinewithIC50valuesunder10µM(1).

Explained that Ag-(I)-NHC complexes induced HL60 colon cancer cell death independent of the caspasecascadeviathemitochondrialAIFpathway(2).

According to a study published in 2003; the researchers identified that a Ag (I)-NHC complex showedsignificantantiproliferativeactivitywithinhibitedthioredoxinreductase.Theinhibitionofthisselenoenzymeidentifiedwithalterationofthecellularredoxenvironment(3).

Inawork;determinedthattheapoptosismechanismofnewAg(I)-NHCcomplexesonlungcancercell(A549).In the result identified thatwhileAg complexeshaving the IC50 level7.1±0.78, cisplatinwas IC50valueof6.59±0.63.AgcomplexesinhibitedthegrowthofA549cancercellsbyinducingG2/Mphasecellcyclearrestand apoptosis. Ag complexes induced apoptosiswith the levels of intracellular ROS. Also these complexesdisruptedthemitochondrialmembranepotential. Further; inducedthecaspase-3activationand ledtothetranslocationofapoptosisinducingfactorandendonucleaseGtothenucleus(4).

Reported that 4-Alkylated Ag-NHC complex exhibited cytotoxic effects in leukemia cells with IC50 valuesof27μM(5).


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PP-26

CYTOTOXICEFFECTSOFNON-HEMOLYTICSKIN-PAROTOIDGLANDSECRETIONSOFTHEBUFONIDTOADSFROMTURKEY

AyşeNalbantsoy¹,MertKarış²,BayramGöçmen²

¹|EgeUniversity,FacultyofEngineering,DepartmentofBioengineering,35100Bornova,İzmir,Turkey.

²|EgeUniversity,FacultyofScience,DepartmentofBiology,ZoologySection,35100Bornova,İzmir,Turkey.Object: Skin-parotoid secretions of amphibians contain a large number of biologically active compoundswhicharethoughttoplayseveralroles,eitherintheregulationofthephysiologicalfunctionsoftheskinorindefense mechanisms against predators or microorganisms. Especially, the biodiversity of biochemicalcompoundsintheauricularandskinglandsoftoadsmakesthemuniquesourcesfornewtherapeuticagents.Material andMethod: Common Toad-Bufo bufo, Caucasian Toad-Bufo verrucosissimus andVariableGreenToad-Bufotes variabiliswere collected in field, then skin secretions obtained by stimulator,while parotoidglandsecretionsobtainedbymanualcompressing.Eachindividualwasrinsedwithultra-purewaterintothetubes, then snap-frozen by liquid nitrogen and then lyophilized. Protein content was determined by BCAassaykit.CytotoxiceffectswasdeterminedagainstHeLa,A549,Caco-2,MPanc-96,PC-3,MDA-MB-231cancercellsandHEK-293asanon-cancerouscell linebyMTTassay.Parthenolidewasusedasapositivecytotoxiccontrol agent. Percentagesof surviving cells and IC50values ineach cellswere calculatedafter incubationwithsecretionsusingGraphPadPrism5.Hemolyticactivityofsecretionswasalsodeterminedonrabbitred-bloodcells.Results:ProteinconcentrationsofB.bufo,B.verrucosissimusandB.variabilissecretionswerecalculatedas3100 µg/ml, 3300 µg/ml, 3480 µg/ml, respectively. Crude skin-parotoid gland secretions of all taxa wereshowed strong cytotoxiceffectonall cancerandnon-cancerous cellswithan IC50values varyingbetween<0.1-6.02μg/ml.Nohemolyticactivitiesatconcentrationsbetween0.5-50μg/mlwereobserved.Conclusion:Furtherinvestigationsneedtofocusontopurifytheactivecomponentsfromtheseskin-parotoidsecretions and determine the possiblemodeof action of secretion-induced cytotoxicity to obtain a betterunderstandingoftheirpotentialuseasanticanceragents. Keywords:Toad,skin-parotoidglandsecretion,cytotoxicity,hemolyticactivity.


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016

PP-27

MICRORNA376FAMILYANDCANCER

YunusAkkoç¹,KumsalAyseTekirdağ¹,AsiyeIşınDoğanEkici²,DevrimGözüaçık¹

¹|DepartmentofMolecularBiologyGeneticsandBioengineering,FacultyofEngineeringandNaturalScience,SabancıUniversity,34956İstanbul,Turkey

²|DepartmentofPathology,YeditepeUniversitySchoolofMedicine,Atasehir,34755Istanbul,Turkey Object: Autophagy, is one of the most well known catabolic processes whose activation can degradeaccumulated proteins as well as damaged organelles for maintaining cellular homeostasis. Beside this,autophagywasfoundtobeassociatedwithseveralabnormalitiesincludingcancerandmetabolicdiseases.Inaddition,miRNAshavebeen implicated inseveral fundamentalbiologicalprocesses includingdevelopment,differentiation,apoptosisandstemcellmaintenance.Moreover,evidencealsosuggeststhatmiRNAsplayarole in cellular transformation and carcinogenesis. Thus, understanding the regulation of autophagicmechanismsthroughmiRNAsmighthavetremendousimportanceinthefieldofcancer.MaterialandMethodOverexpressionofMIR376BinMCF-7cellshasbeenutilizedandseveralmonoclonecellspickedandculturedunderselectioncondition.Forfurtheranalysis,monocloneswereevaluatedbytheirautophagiccapacityviaLC3shift,p62accumulationandmiR-376btargetproteinstatus.Afterthecharacterizationofclones,severalgrowthanalyseswereperformedeithershortorlongtermassaysinvitro.ClonogenicpotentialofMIR376Bstablyoverexpressingandcontrolcellswereanalyzedbytheirabilitytocreatecoloniesbycolonyformationassay.On theotherhand,Gamma-H2AX foci analysis andROSmeasurementbyDCFDAwas carriedout toidentifytheDNAdamageandoxidativestress,respectively.Result:Asaconsequenceofautophagyderegulation,accumulationofp62wasobservedinmiR-376bstablecells. Intriguingly, intracellular ROS levelwas also increased and accumulationof ROS localized around themitochondria. Inaddition tosusceptibilityofoxidativestress, lossofautophagymakescellsmoreprone toDNAdamage.Although inshort termassays,growthattenuationofmiR-376bstablecellswasobserved; incolonyformationassay,thosecellsformedmoreandbiggercolonies.Conclusion: We identified for the first time that MIR376B as a key miRNA which might has a role intumorigenesisinbreastcancer.*Thisworkwas supportedbyTheScientific andTechnologicalResearchCouncilofTurkey (TUBITAK)1001-114Z982GrantandSabanciUniversity.Keywords:Macroautophagy,mammalianautophagyregulation,microRNA,MIR376B,cancer


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016

PP-28

THECYTOTOXICEFFECTSOFETOPOSIDE,PYCNOGENOL®ANDTHEIRCOMBINATIONONA549CELLLINE

A.CansuKilit¹,EsraAydemir¹

¹|AkdenizUniversity,ScienceFaculty,BiologyDepartment,Konyaaltı,Antalya

Object: Chemotherapy is the most common method used in treating cancers and directly targets ateliminatingcancercellsusingchemicalswithcytotoxiceffect.Becausethetargetofchemotherapyistumorcell,theagentisusedinhighdosesduringapplicationandthiscreatestoxicresponsescausingsideeffectsonnormal cells. Hence chemotherapy is combinedwith some cancer treatments to increase the therapeuticefficacy by reducing side effects. Etoposide belongs to the class of drugs known as podophyllotoxinderivatives.AtopoisomeraseIIinhibitorwithinetaposidepromotesapoptosisincancercellsandithasbeenusedforthetreatmentof lungcancer.Pycnogenol® isakindof flavonoidand itcanbeusedasthedietarysupplement.It isrichofmanyphytochemicalsofmedicalvalue, isextractedfromtheFrenchmaritimepinebarkPinuspinaster.InthisstudythecytotoxiceffectsofEtoposideandPycnogenolcombinationonA549cellswasresearched.MaterialandMethod:A549cellswereculturedinRPMI1640mediumcontaining10%fetalbovineserum,10µg/ml gentamicin and 5% sodium pyruvate. The cells were incubated in 5% CO2 at 37 °C. The cells weretreatedwithvariousconcentrations (500-12,5µg/mL)ofEtoposide,Pycnogenolandtheircombination.Cellviabilitywasmeasured after 24 h treatments. Cytotoxic effects of drugswere investigated usingMTT cellproliferationkit.Results: Etoposide alone caused 34.20%, 27.87% and 24.28% cell death at 500, 400 and 300 µg/mLconcentrations,respectively.IC50valuesforPycnogenolandPycnogenol+Etoposideare485and195µg/mL,respectively.Conclusion:Attheendof24hPycnogenolandEtoposidecombinationsatdifferentdosesexertedsynergisticeffectonA549celldeathKeywords:Etoposide,Pycnogenol®,Cytotoxicity,Combinetherapy


4-7

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016

PP-29

THEAPOPTOTICEFFECTOFBONGARDIACHRYSOGONUMONHUMANGASTRICCANCERCELLLINEHGC-27

EbruTemiz¹,AbdullahTuncayDemiryürek²,SerdarÖztuzcu¹,AhmetSaracaloğlu²,Şeniz

Demiryürek³,BilgeŞener⁴,BeyhanCengiz⁵,MustafaUlaşlı¹,CelaletdinCamcı6

¹|DepartmentofMedicalBiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey²|DepartmentofMedicalPharmacology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,

Turkey³|DepartmentofPhysiology,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

⁴|DepartmentofPharmacognosy,FacultyofPharmacy,GaziUniversity,06330Ankara,Turkey⁵|DepartmentofMedicalGenetics,FacultyofMedicine,GaziUniversity,06560Ankara,Turkey

6|DivisionofMedicalOncology,DepartmentofInternalMedicine,FacultyofMedicine,UniversityofGaziantep,27310Gaziantep,Turkey

Objective:Despitethegreatprogressinthetreatmentofgastriccancer(GC),itisstillthethirdleadingcauseofcancerdeathworldwide.GCstillcarriesapoorprognosis.Therefore,itisimperativetoidentifynovel,lowtoxicandeffectiveanticancerdrugs.Inthisstudy,weinvestigatedtheapoptoticactivityofhexaneextractofthetuberskinofBongardiachrysogonumonthehumanGCcelllineHGC-27.Material andMethod:HGC-27 cellswere cultured inDMEMmedium supplementedwith10% fetal bovineserum.Whencellsreached70%ofconfluence,theyweretreatedwithhexaneextractorDMSOduring24h.ApoptoticcellswerequantifiedbyAnnexin-V/7AAD-positivestaining,usinganAnnexin-V-FITC/7AADkitfromBeckmanCoulter.Positivecellswereanalyzedonaflowcytometer(NAVIOSBeckmanCoulter,USA).Forthecellcycleassay,BDCycletest™PlusDNAReagentkit(Biosciences,Germany)wasused.Cellcycleanalysiswasperformed by using flow cytometer. For gene expression study, mRNA was isolated from cells by usingmiRNeasyMini Kit (Qiagen GmbH, Germany). Then cDNA was produced with the Ipsogen RT Kit (QiagenGmbH).qRT-PCRwasperformedbyRotorGene6000(QiagenGmbH,Hilden,Germany).Westernblotanalysiswasperformedforproteinexpression.Results: Apoptosis of the HGC-27 cells was stimulatedwith hexane extract of the tuber skin (2.8%)whencompared to DMSO (0.5%). Nomarked changes was observed in cell cycle assay (hexane extract: G0/G158.9%,G20.1%,andS41.0%;DMSO:G0/G160.0%,G20.3%,andS39.7%).WefoundthatNFKB,p27,andp53geneandproteinexpressionswerenotsignificantlymodifiedwithhexaneextracttreatment(p>0.05).Conclusion:OurresultsshowedthathexaneextractofthetuberskinofBongardiachrysogonumexhibiteditsanti-tumoractivityagainstHGC-27cellsthroughpromotingapoptosis.ThisstudywassupportedbyaGaziantepUniversityproject(TF.13.10).Keywords:Apoptosis,cellcycle,flowcytometer,gastriccancer,geneexpression


4-7

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016

PP-30

ANALYSISOFCELLDEATHINHCT-116UNDERHYPOXICCONDITION

SevtapGökalp¹,FeyzanÖzdalKurt²,TunaÖnal¹,CananTürkoğlu²,ElginTürközUluer¹,HSedaVatansever1,3

¹|CelalBayarUniversity,FacultyofMedicine,DepartmentofHistologyandEmbryology,Manisa,Turkey

²|CelalBayarUniversity,FacultyofSciences&Letters,DepartmentofBiology,Manisa,Turkey3|NearEastUniversity,ExperimentalHealthScienceResearchCenter,Nicosia,NorthCyprus

Object: Colorectal cancer is a common malignant neoplasm prevalent in both developed and developingcountries.Thisdiseaserankssecondamongcausesofcancer-relateddeathsworldwide,comprising10%–15%of all forms of cancer. Hypoxia is a common feature of the microenvironment of solid tumors. Hypoxia-regulated genes are known to be involved in multiple biological processes, such as proliferation,angiogenesis, metabolism, immortalization, migration, carcinogenesis and apoptosis expression. Apoptosisplayan important role in the involvementofhypoxia. In this study,weaim to identify theanalysesof celldeathinhypoxiaconditiononhumancoloncarcinomacellline.MaterialandMethod:Humancoloncarcinomacellline(HCT-116)waspurchasedfromATCC.Thecellswerecultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine and 1%penicilin-streptomycine 37◦C in a 5% CO2 humidified atmosphere. The cells were passaged after reaching80%monolayer confluencyand separated in twogroups.Group1 cellswere cultured innormal condition,Group2cellswereculturedinhypoxiachamberwhichhaveagasmixtureof5%CO2,3%O2and92%N2for36h.Theywerethenfixedwith4%paraformaldydeandcelldeathanalyseswereperformedwithTUNEL.Thedistribution of anti-Bax, anti-Bcl2, anti-caspase 3 and anti-cytochrome-c were investigated using indirectimmunoperoxidasestaining.ResultsTUNELpositivecellsweredetectedinbothgroup1andgroup2.Ingroup1, thedistributionofcaspase3,Bcl2andcytochrome-cwereobservedasaweakstaining, theyweremoredetectableingroup2.Inaddition,positiveimmunoreactivityofBaxwasdetectedonlygroup2.Conclusion:ThesensitivityofHCT-116cells isassociatedwiththeupregulationofBcl-2,Bax,caspase-3andcytochrome-cactivitiesinhypoxiccondition.Thehypoxiamaytriggertheexpressionofanti-apoptoticproteinbcl-2topreventthecelldeath,andalso,intrinsicapoptoticpathwaywasinducedtocontrollingofcelldeathKeywords:Coloncarcinoma,hypoxia,apoptosis


4-7

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PP-31

INVESTIGATIONOFTHEMDM2GENEPOLYMORPHISMINPATIENTSWITHACUTEMYELOIDLEUKEMIA

CerenTekin,SibelBayılOğuzkan,MehmetÖzaslan,HandanHaydaroğluBaş,SelinBüdeyri,

MustafaPehlivan

Aim : Leukemia ismalign diseasewhich originated from bonemarrow lymphatic and hematopoietic stemcells.Acutemyeloid leukemia isaclassof leukemiawhich is shownthatbothofphenotypicandgenotypicheterogeneity.Thisdiseaseisdescriptedoverall100cytogeneticaberattionandvarygenesmutation.MDM2 (Murinedoubleminute 2) gen is a proto-oncogen and MDM2 gen polymorphisms which studiedsomedifferentcancertypesandthisisassociatedwithlotsofcancertypes.Method:This study is researchof the relationshipbetweenoccuringMDM2gen354A/Gand (MboII) -410T\G(SNP309)singlenucleotidpolymorphismsandAML.MDM2genpolymorphismsoneofthemis354A\GregionwhichAdeninenucleotideconverttoguaninenucleotideandtheotheroneis-410T/GregionwhichTiminnucleotideconverttoguaninenucleotidearedeterminedinthisstudy.Result: For that reason80AMLpatientswhohavediagnostedAMLand20healthypeopleareused in thisstudy.DNAisisolatedfromAMLpatientsandcontrolgroup’sblood.BothoftheseregionareworkedbyRT-PCR.Dataanalysesof354dA/GpolymorphismisdeterminethatallofthepeoplewhoareincluededinthisstudyhaveAAgenotype.Conclusion:ComparedofMDM2gen354A/Gpolymorphismsbetweenpatientandcontrolgroupshavenotfoundmeaningfulby istatistically (p<0,005). -410T\Gregionpolymorphismresult is foundthat the19wildtype (TT) of 80 AML patients, 25 heterozygote genotype (TG) AND 36 patients are mutant type (GG)genotype.AttheendofthestatisticalanalysisofMDM2gen-410T\Gpolymorphismshasfoundstatisticallysignificantbetweentwogroups(p˂0,05). Keywords:MDM2,AcuteMyeloidLeukemia,RT-PCR


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PP-32

ACTIVATIONOFENDOPLASMICRETICULUMSTRESSMIGHTNOTBESUFFICIENTTOINDUCEAPOPTOSISINMETASTATICBREASTCANCER:POSSIBLEROLECYTOPLASMIC

EXPRESSIONOFCHOP.

SayraDilmaç¹,NurayErin²,GamzeTanrıöver¹'²

¹|AkdenizUniversity,FacultyofMedicine,DepartmentofHistologyandEmbryology²|AkdenizUniversity,FacultyofMedicine,DepartmentofMedicalPharmacology

Object: Endoplasmic reticulum (ER) is the principal organelle responsible for multiple cellular functionsincludingproteinfolding,maturationandthemaintenanceofcellularhomeostasis(1).ERstressispartoftheunfolded protein response (UPR) which is involved in cell death(2).Markers of ER stres include GRP78, aproteinkinase,CHOPandp-PERKwhichundernormalconditionsactivatesapoptoticpathwayandmayhaveanti-tumoraleffects(2-3).Therefore,thegoalofthepresentstudyistodeterminetheexpressionpatternsofERstressproteinsinmetastaticbreastcancer.Material and Method: We previously isolated liver (4TLM) and heart (4THM) metastatic as well as non-metastatic67NRcellsformedby4THMmurinebreastcarcinomausinganorthotopicmodel(4).4TLM,4THM(100.000 cells/mouse) and 67NR (1.000.000 cells /mouse) cells were inoculated into the right uppermammarypadof8-10weeksoldfemaleBalb-cmice.Necropsieswereperformed25-27daysafterinjection.p-PERK, GRP78, CHOP, Bcl-2 immunoreactivities were examined in primary tumors by usingimmunohistochemistry.ImmunoreactionintensitywasquantifiedusingimageJsoftware.Results:WeobservedthesignsofERstresssuchthatp-PERK,GRP78expressionsweresignificantlyincreasedinhighlymetastatic4TLMprimary tumor tissues compared tonon-metastatic tumors.DifferentlyER stressprotein CHOP which supposed to be nuclear under stress conditions were localized in cytoplasm(5). It isknown that cytoplasmicCHOP in thepresenceofBcl-2expressionpreventsapoptotic cell deathdue toERstress.InterestinglyCHOPandBcl-2immunoreactivitywashigherinnon-metastatictumorgroups.Conclusion: These results demonstrated that ER stress increased in more aggressive metastatic tumors(4TLM) and expression of cytoplasmic CHOP might counteract these apoptotic signals contributing toaggressivenessof4TLMtumors.Keywords:ERstress,breastcancer,CHOP,Bcl2,immunohistochemistry


4-7

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016

PP-33

CHARACTERIZATIONOFTHEEFFECTOFSEVERALETODOLACDERIVATIVESONCANCERCELLLINES

SevgiKoçyiğit¹,PınarMegaTiber¹,PelinÇıkla-Süzgün²,Ş.GünizKüçükgüzel²,OyaOrun¹

¹|DepartmentofBiophysics,FacultyofMedicine,MarmaraUniversity

²|DepartmentofPharmaceuticalChemistry,FacultyofPharmacy,MarmaraUniversityObject Etodolac is a nonsteroidal anti-inflammatory drug with analgesic and antipyretic properties. It hasinhibitoryeffectsoncyclooxygenase-2(COX-2)activationandsimilartootherCOX-2inhibitors,itshowsanti-tumorigenic effects.We studied anti-proliferative effects of hydrazone and triazole derivatives of etodolac(SGK206andSGK242,respectively)synthesizedbyDr.Kucukguzel[1]ontwobreastcancercelllines,MCF-7andMDA-MB-231.MaterialandMethodsUsing theMTTcolorimetricmethod,derivativesofetodolacwereevaluated invitroagainstthebreastcancercelllines,forcellviabilityandgrowthinhibitionatdifferentdosesfollowing24,48,72 and 96 hours incubations. Apoptosis was also evaluated in both cell lines after treatment in variousconcentrationsusingtheTaliimage-basedcytometer.Inaddition,PGE2concentrationsweremeasuredintheconditionedmediausingPGE2ELISAkit(Enzo).Results: We first checked Cox-inhibiting activity of the drugs with PGE2 assay. All drugs inhibited PGE2release,butderivativesaremoreefficientinlowconcentrations.MTTresultsindicatedthatetodolacdoesn’thavetoxicityonthecells,evenafterlongincubationperiodsin100µMconcentration.Ourderivatives,ontheotherhand,hadsubstantialeffectinmuchlowerdoses(10µM).EspeciallySGK-206showedaregulardose-response curve, with an IC50 value of 32 µM against the MCF-7 and 38 µM against the MDA-MB-231.CorrespondingvaluesforSGK-242were12µMand19µM,respectively.Higherconcentrationsshowedtoxiceffectsinbothdrugs(>25µM).Apoptosisalsoincreasedinbothcelllinesinadose-dependentmanner.Conclusion:Our results show thatderivativesofetodolacweremoreeffectiveboth inPGE2 inhibitionandcytotoxicity.Similarlyapoptoticeffectofetodolacwasverylowcomparedtotheothercompounds.Wearegoingtoextendourstudiesonthesearchoflow-doseeffects(<25µM)ofthesedrugsindifferentcancercelllines.Keywords:etadolac,breastcancer,apoptosis,cellcytotoxicity.


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PP-34

ANTI-CYTOTOXICANDANTI-GENOTOXICEFFECTSOFCONJUGATEDLINOLEICACIDANDWHEYPROTEININACROLEIN-TREATEDRATS

VedatŞekeroğlu¹,ZülalAtlıŞekeroğlu¹,BirsenAydınKılıç²

¹|OrduUniversity

²|AmasyaUniversity

Object:Ourstudyaimedtodeterminetheeffectsofconjugatedlinoleicacid(CLA)orwheyprotein(WP)onthefrequencyofmicronucleus(MN)andtheratioofpolychromaticerythrocytes(PCEs)againstacrolein(AC)-inducedtoxicityinrats.Material and Method: Thirty-six healthy rats were obtained from the Experimental Animal Center ofUniversity of OndokuzMayıs in Samsun, Turkey. The studywas approved by theMedical Research EthicsCommittee(HADYEK2014/24)oftheuniversity.AnimalswereorallygavagedwithCLA(200mg/kg/day),WP(200mg/kg/day),AC(5mg/kg/day),CLA+AC(200+5mg/kg/day)andWP+AC(200+5mg/kg/day)sixdaysperweekfor30days.Results:ItwasfoundthatACsignificantlyincreasedtheformationofMNinbonemarrow.ItalsosignificantlydecreasedtheratioofPCEs.NosignificantdifferencesinMNandPCEsvalueswereobservedintheanimalsreceived the CLA or WP alone compared to the control group. Co-treatment with CLA+AC or WP+ACsignificantly increased the ratioofPCEs.Although the co-treatmentwithCLA+ACorWP+ACdecreased theformationofMN,thedecreaseswerenotfoundsignificantwhencomparedtocontrolgroup.Conclusion:OurresultsindicatethatCLAandWHmayplayabeneficialroleinreducingthecytotoxicityandgenotoxicityinducedbyACinrats.

Keywords:Acrolein;Wheyprotein;Conjugatedlinoleicacid;Cytotoxicity;Genotoxicity


4-7

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PP-35

PISTACIAEURYCARPAINHIBITSCELLPROLIFERATIONANDINDUCESAPOPTOSISINCOLORECTALCANCERBYMODULATINGAPOPTOTICPATHWAYGENES

MehmetKadirErdoğan¹,CanAliAğca²,HakanAşkın²

¹|BingölUniversity

²|AtatürkUniversityObjectColorectalcancer(CRC)isthethirdmostcommoncancer.Itsglobalincidenceandmortalityhavebeenon the rise. Recent strategy of therapies has involved the use of non-steroid anti-inflammatory drugs andcyclooxygenase-selective inhibitors [1]. The genus Pistacia consists of small trees of the cashewnut familyAnacardiaceaeandisnativetotropicalandsubtropicalAsiawhereitsmembershavelongbeencultivatedfora variety of uses. The trunk of Pistacia species produces a characteristic exudate called mastic gum. Themastic gum and oil are medicinally used against rabies, snake bites, baldness, scabies, as well as inprescriptions for stomach, intestine, bladder, and liver inflammations, oral and dental diseases [2]. In thisstudy, growth-inhibitingandpro-apoptoticeffectsofhexane, chloroformandmethanolextractsofPistaciaeurycarpainHT-29colorectalcancercelllinewereinvestigated.MaterialandMethodAerialpartsofPistaciaeurycarpawerecollectedinBingölprovince.Hexane,chloroformandmethanolextractionweredonebySoxhletextractor.DoseandtimedependentcytotoxicandapoptoticeffectsofPistaciaeurycarpawereevaluatedbyMTTCellProliferationKitandCellDeathDetectionElisaKit,respectively(RocheDiagnostics,Germany).Manufecturer’sprotocolwasfollowedforanalyses.Combinationof 5-FU andPistacia eurycarpawere also applied toHT-29 colorectal cell line for detecting the synergism.pTEN,AKT,MAPK,mTOR,VEGFReceptor2,p53andβ-actingeneexpression levelsweremeasuredbyRT-PCR.WesternblotanalyzewereusedtodeterminepTEN,AKT,MAPK,mTOR,VEGFReceptor2,p53andβ-actinproteinlevels.Results According to Cell viability rates, gene and protein expression levels results, there is a synergismbetweenPistaciaeurycarpaand5-FU.ConclusionInconclusion,Pistaciaeurycarpaextractsrepresentsapotentialsourceforanti-proliferativeandapoptoticagentsincombatingCRC.Keywords:Colorectalcancer,Pistacia,extract,MTT,p53


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016

PP-36

ANTI-CANCERACITIVITYOFAHYDRAZONEDERIVATIVEOFETODOLACINK562LEUKEMIACELLLINES

PelinYonarKaplan¹,SevgiKoçyiğit¹,PınarMegaTiber¹,PelinÇıklaSüzgün²,Ş.GünizKüçükgüzel²,

OyaOrun¹

¹|DepartmentOfBiophysics,SchoolOfMedicine,MarmaraUniversity,İstanbul,Turkey²|DepartmentOfPharmaceuticalChemistry,SchoolOfMedicine,MarmaraUniversity,İstanbul,Turkey

Object: Etodolac is a nonsteroidal anti-inflammatory drug that selectively inhibits Cox-2 enzyme. Cox-2inhibitors have been reported to have anti-tumorigenic effects on cancer cells through inhibition ofproliferation and induction of apoptosis. Hydrazones constitute an important class of compounds for newdrugdevelopment.Studiesimpliesthathydrazide-hydrazonederivativesofanti-cancerdrugscouldbemoreeffectiveandlesstoxic.Here,westudiedanti-proliferativeeffectsofahydrazonederivativeofetodolac(SGK205)on leukemiacellline.HerewereportitseffectsonmyelogenousleukemiacelllineK562.MaterialandMethods:Anti-proliferativeeffectsofthedrugwasdeterminedusingMTTcolorimetricmethod,atdifferentdosesfollowing24and48hourincubations.Apoptosiswasevaluatedbytwodifferentmethods,wherecellswereeitherstainedwithAnnexin-V/PIandanalyzedusingtheTaliimage-basedcytometerorwithJC-1probe,whichwasusedtodetectmitochondrialmembranepotentialchanges.Results:Therewasadose-dependentdecreaseinproliferationdetectedbyMTTassayandIC50valueswere21.3and17.7µMattheendof24and48hourincubations,respectively.Apoptosisassaysalsodisplayedadose-dependentincreaseuntil40µMofthedrug.Afterthisconcentrationdrugwashighlytoxicforthecells.Etodolacdidn’tshowneitheranti-proliferativenorapoptoticeffectsatindicateddosesandincubationtimes.Conclusion: Several etodolachydrazonederivativeswere synthesizedbyDr.Küçükgüzel et al. and cytotoxiceffect of compound SGK-205was tested at a concentration of 10 µM against the full panel of 60 humancancercelllinesbytheNationalCancerInstitute(NCI).Thesepreliminaryresultsdisplayedastrongeffectonleukemiacelllines.Ourfurthercharacterizationofthecompound,thisstudyshowedthatSGK-205hadindeedanti-proliferativeand apoptotic effects on K562 cells in a dose-dependent manner, but concentrations higher than 40 µMresultsintoxicity.


4-7

May2

016

PP-37

MYCINHIBITIONMAYBEAPROMISINGTHERAPEUTICSTRATEGYFORASIGNIFICANTFRACTIONOFSCLC

OnurTokgün¹,PervinElvanTokgün¹,HakanAkça¹,GülserenBağcı¹

¹|PamukkaleUniversity,SchoolofMedicine,DepartmentofMedicalBiology

Object:Smallcelllungcancer(SCLC)isthemostaggressivetypeoflungcancerwithhighmortality.OneoftheMYCfamilygenes,MYC,MYCL1orMYCN,isamplifiedandoverexpressedin~20%ofSCLC;therefore,wehaveselectedtheseMYCfamilygenesasstrongcandidatesoftargetsfortherapyforSCLC,andare investigatingthebiologicalsignificanceofMYCfamilygeneamplificationforSCLCcellsbyeitherdown-regulationofMYCfamilygenesorsuppressionoftheiractivitieswithinducibleshRNAsandsiRNAs.Material andMethod: SCLC cell lines were used in this study. To check the effect of Myc expression oncellular proliferation we designed constitutive Myc expression vector by using pCDH-CMV vector as abackbone.After constitutivevector construction,weplanned to constructa vector containingan induciblesystemfortheexpressionofc-MycbytransferingMYCcDNAclonedintopCDHvectorintopTRIPZvector.Inthesameway,aconstitutivesystemforknockdownofc-Mycbyshorthairpinexpressionisnotsuitablefortheevaluationtheeffectofc-Mycexpressionbyexpressionprofiling.Therefore,weplannedtoconstructaninduciblesystemforknockdownofMYCusingpTRIPZ.TheeffectofMycexpressiononcellularproliferationandgeneexpressionprofilingwereevaluatedinMycoverexpressedandknocdownSCLCcells.Results: Constitutive and inducible expression of Myc induced cellular proliferation in Myc non-amplifiedSCLC cells. Importantly, c-Myc downregulationmarkedly reduced the enhanced proliferation of SCLC cellswithectopicexpressionofc-myc.AndinMYCoverexpressedcellsweevaluatedinhibitoryeffectsofshRNAsoncellularproliferation.DuetochangesinMycexpression,weevaluatedthechangesofsomeproliferationandapoptosismarkersbywesternblotandqRT-PCR.Conclusion:TheresultsindicatethatSCLCcellsareaddictedtoMycproteinsfortheirgrowthand,therefore,arehighlysensitivetoMYCinhibition. Keywords:SCLC,MYC,shRNA,apoptosis,proliferation


4-7

May2

016

PP-38

EFFECTSOFBOROTUNGSTATEAGENTONCELLSURVIVALANDMORPHOLOGYINNONSMALLLUNGCANCERCELLLINE

PınarÖztopçuVatan¹,HiraUğurlugüngör¹,EbruKanar²,AsımOlgun²,AlperTolgaÇolak²

¹|EskisehirOsmangaziUniversity

²|DumlupınarUniversity

Object:Despiteofclinicaltreatments,lungcarcinomaisoneofthemostcommoncausesofdeathbywayofcancer. Recent studies demonstrated that polyoxometalates (POMs) exhibit potent antitumor activity.Borotungstate(K16[Ni(H2O)6]2[BW12O40]4.48H2O)isaPOM.It’sacompoundcontainingboronatomwhichwas previously synthesized in our lab. Since the function of borotungstate has not been defined yet, wedecided to test the possible proliferative andmorphological effect on human. non small cell lung (H460)carcinoma.Material and Method: The cells were inoculated at a concentration of 1x104 cells/well. Then they weretreatedwith10,25,50,75and100µMborotungstatefor24or48hr.Attheendofincubationperiods,cellsurvivalwasmeasuredwithMTTassay.Inthisstudyweused10,25,50,75and100µMdosesofcarboplatinaspositive control for24and48hr. Furthermore, the cellswere treatedwith10 to100µMborotungstatedosesfor24hrandmorphologicalchangesweredetectedunderaninvertedlightmicroscope.Results:BorotungstatedecreasedthesurvivalofH460cellsinadoseandtimedependentmanner.Thehalfmaximalinhibitoryconcentrationvaluesofborotungstateweredeterminedas51µMfor24hrand31µMfor48 hr respectively. Borotungstate treatment of 25 µM decreased cell counts 50, 75, and 100 µM doseschangedthecellmorphologyanddeclinedthecellcounts.Conclusion: In thisstudyanewsyntheticborotungstateagent,examined for the first time inahumannonsmalllungcancercellsandobtainedsomepreliminaryresultsaboutthecytotoxicandmorphologicaleffectsofthisagent

Keywords:Borotungstate,MTT,cellmorphology,lungcarcinoma


4-7

May2

016

PP-39

APOPTOTICEFFECTOFFERULICACIDONDU145PROSTATECANCERCELLS

ÖnderYumrutaş²,EsraBozgeyik¹,İbrahimBozgeyik²,EbruTemiz¹,PınarYumrutaş¹,SerdarÖztuzcu¹

¹|GaziantepÜniversitesi²|AdıyamanÜniversitesi

Object: It is known that phenolic compounds are important phtochemicals to have the strong activities.However, information about their anticancer activity including cell cytotoxicity, antiproliferation andapoptotic effects on cancer cells is limited. Hence, in present study, we aim to determine of anticanceractivityofferulicacid(FA),whichissynthesizednaturallyinplants,onprostatecancercellsDU145.Material andmethod:MTT testwasused todetermine theantiproliferativeandcytotoxiceffectsof FAonDU145cells.0.2,1,5,25µg/mldosesofFAwereusedforapplication.AftertheapplicationofdosesofFA,apoptosis were determined by using a FACS with staining Annexine V and Propodium iodide (PI). Then,apoptotic genes; caspase 9, caspase3 and Bax, and antiapoptotic genes; Bcl-2 and Bcl-Xl expression levelsweredeterminedbyRealTimePCR.Results:Accordingtoourresults,cytotoxiceffectonDU145cellwasonlyobserved in25µg/mldoseofFA.However,anyoftheapplicationdosesshowedanapoptoticeffectonDU145cells.Theyexhibitedanecroticeffectratherthantheapoptoticeffect.Althoughalterationsinexpressionlevelsofcaspase9,caspase3,Bax,Bcl-2andBcl-Xlgenesweredetermined,thisalterationswasstatisticallyinsignificant.Conclusion:Consequently,wecansaythatFAhavenotaapoptoticeffectonDU145prostatecancercelllines.Keywords:Ferulicacid,antiproliferation,apoptosis,Caspas,Bcl-2,


4-7

May2

016

PP-40

CANARZANOL,APOTENTMPGES-1INHIBITOR,USEASATARGETFORPRIMARYORMETASTATICBREASTCANCERCELLS?

MertPak¹,RemziyeKendirci¹,HilalKabadayı¹,H.SedaVatansever¹'²

¹|CelalBayarUniversity,FacutyofMedicine,DepartmentofHistologyandEmbryology,Manisa,Turkey

²|NearEastUniversity,ExperimentalHealthScienceResearchCenter,Nicosia,NorthCyprus

Arzanol is a prenylated heterodimeric phloroglucinyl α-pyrone chemically known as 3-(3-acetyl-2,4,6-trihydroxy-5-(3-methylbut-2-en-1-yl)benzyl)-6-ethyl-4-hydroxy-5-methyl-2H-pyran-2-one. Prostaglandin E2(PGE2)isabioactivelipidthatcanelicitawiderangeofbiologicaleffectsassociatedwithinflammationandcancer. Inourstudy,theeffectsofArzanolonprimaryandmetastaticbreastcancercell linesviacaspase-3expressionswere investigated.Humanprimary (MCF-7)andmetastatic (M4A4)breastadenocarcinomacelllinewere cultured in RMPI-1640 containing 10% FCS, 1% L-glutamine and 1% penicillin-streptomycin until80%ofconfluence.Cellswereseparatedintwogroups;controlandArzanoltreatedgroups.ControlgroupofMCF-7andM4A4cellswereculturedwithnormalculturemedium. InArzanol treatedgroupofMCF-7andM4A4wereculturedwithdifferentdosageofArzanol (0.5µg/ml,10µg/ml,20µg/mland40µg/ml) for96hours.CellcultureswerecollectedforLDHassay.Afterfixationofcellswith4%paraformaldeyde,distributionof caspase-3 was detected with indirect immunoperoxidase techniques. After LDH analyses, Arzanol wasmorecytotoxic inMCF-7 cellswhencomparewithM4A4cells. Inaddition, toxicitywashigher in40µg/mldosage of Arzanol.Moderate or strong caspase-3 immunoreactivitywas detected all dosage of Arzanol inM4A4 cells. In control group ofMCF-7 andM4A4, caspase-3 immunoreactivitywas negative. Arzanolwasinduce caspase-3 secretion in M4A4 cells, metastatic breast adenocarcinoma cells, than MCF-7, primaryadenocarcinoma cell line. In addition, Arzanol was cytotoxic for MCF-7 cells, therefore weak caspase-3immunoreactivitywasobserved.Keywords:Breastcarcinoma,Arnazol,caspase-3


4-7

May2

016

PP-41

THEEFFECTEDGENESANDPATHWAYSBYUSNICACIDINSIDETHECELL

ZehraDilşadÇoban¹,HalideDemir,TunaKaraer,BüşraAtmaca,ŞefikGüran¹

¹|GülhaneAskeriTıpAkademisiTıbbiBiyolojiAD.

Object:Usnicacid(UA)isadibenzofuranderivativemetabolitefoundinlichen.Ithasanticancerpropertiesinmanycancerssuchascolon,lung,breastcancerandleukemia.Butthemechanismsofanticancereffectisstillunknown. Singh at all showed that UA stopped the growth of A549 lung cancer cells andmade them toundergo apoptosis. In our study,we tried to find outwhichmoleculeswere effected during apopitosis byusing UA on A549 cells. So we could explain the intracellular mechanisms of UA; thought to be an idealantitumoralagent,onmolecularbasisforthefirsttimeintheliterature.MaterialsandMethods:A549cellswereobtainedfromATCC.ToxicdoseoftheUAisdeterminedusingthe"MTT cell proliferation assay" method. The cultured cells were used for RNA isolation and than cDNAsynthesis.ThegeneexpressionanalysesweredoneforProcaspase3,Bcl-xLandBAXgenesbyRT-PCR.Results:AccordingtoMTTresults,thedoseofUAthatweusedinourstudy(50-100µM)wasdeterminedtobenon-toxic.Inourstudy,Procaspase3geneexpressionwasfoundas0,003inthecontrolgroupanditwasfound same as 0.003 in the 50 µM concentration group and 100 µM concentration group. The geneexpressionlevelsofBcl-XLwerefoundas0.19incontrolgroup,0.55inthe50µMconcentrationgroupand0.2 in100µMconcentrationgroup. IncontrolgroupBAXgeneexpressionwasfoundas0.98 inthecontrolgroupwhileitwasfound0.64inthe50µMconcentrationgroupand1.57inthe100µMconcentrationgroup.Conclusion:Accordingtotheresults,thedosesofUAthatweusedinourstudy,hasnotoxiceffectonA549cells.Accordingtothegeneexpressionanalysis,nochangewasshowedingeneexpressionofProcaspase3between three groups. Bcl-XL gene expression increased in the 50 µM concentration group. The geneexpression levels were found similar in the 100 µM concentration group and control group. BAX geneexpression decreased in the 50 µM concentration group compared to the control group and increased inthe100µMconcentrationgroup.Consequently,whileUAwaseffectiveonthegenesofthefinalpathwayofapoptosis (Bcl-XL and BAX genes), was not effective on Procaspase 3 gene known as involved in themitochondrialpathwayofapoptosis.Our findings indicate thatUA iseffective throughapoptoticpathwayswithinthecell. Keywords:Usnikasit,akciğerkanseri,apopitoz,genekspresyonu


4-7

May2

016

PP-42

THERAPEUTICAPPROACHESFORPANCREATICCANCERSTEMCELLS

GülinnazErcan¹'²,AyferKarlıtepe¹,BülentÖzpolat3

¹|EgeUniversity,MedicalFacultyDepartmentofMedicalBiochemistry,Izmir,Turkey²|EgeUniversity,InstituteofHealthSciencesDepartmentofStemCell,Izmir,Turkey

3|UniversityofTexas,MDAndersonCancerCenter,Houston,Texas,USA

Pancreatic cancer (PaCa) is cur¬rently the fourth most frequent cause of can¬cer-related deaths in theUS.Pancreatic Ductal Adenocarcinoma (PDAC), the major histological subtype comprising 90% of allpancreatic cancers, displays local invasion andmetastasis during early stages of the diseasevia developingintrinsicresistancetomostterapeutics,contributingtoitsnotoriouslypoorprognosis(~6monhtsofmediansurvival with 1-5% 5-year survival rates). PDAC is a highly complex malignancy due to many molecularalterations, including Mutated KRAS (~90% of the cases ), TP53, TGF-β, hedgehog and Wnt signalingpathways.Giventhatcancerstemcells(CSCs)haveacrucialrolenotonlyintumorinitiationandprogressionbutalsorecurrenceofthedisease,theyareproposedtobeexcellenttargetsforeffectivenoveltherapeuticapproaches. Here, we presented the recent therapeutic strategies targeting pancreas CSCs such aschemotherapeuticdrugs,miRNAs,immunotherapyandparticularlynaturalcompounds


4-7

May2

016

PP-43

THEEFFECTSOFALPHALIPOICACIDASAPOWERFULANTIOXIDANTINBUSULFANTREATMENTFORCHRONICMYELOIDLEUKEMIA

MerveAksoy¹,PınarErçetin²,ÇetinPekçetin¹,AyçaPamukoğluKaynar²,SafiyeAktaş²

¹|DokuzEylülÜniversitesiTıpFakültesiTemelTıpBilimleriHistolojiveEmbriyolojiAnabilimDalı

²|DokuzEylulUniversityInstituteofOncologyDepartmentofBasicOncology

Purpose:Chronicmyeloid leukemia(CML)isamyeloproliferativediseasecausing leukocytosis inpheriphericbloodwhich is characterizedbyuncontrolledclonalproliferationanddecreasedapoptosisofmyeloid stemcells.BusulfanhasbeenusedforCMLasaconventionalchemotherapy.Alphalipoicasid(ALA);istheuniqueantioxidantthatissolublebothinlipidandwater.ALAisnamedasaglobalantioxidantbecauseofitsroleinpreventingofradicalmetaboliteproduction,clearingoffreeradicals,repairingofdamagecells,inhibitionofchain reactionswhichproduce secondary radicals and increaseendogenousantioxidant capacity.Busulfan,hasseveralsideeffectssuchastestisdamageandALAisacandidateprotectiveagentforthissideeffect.TheaimofthisstudyistosearchtheeffectofALA,bothaloneandincombinationwithbusulfanonK562CMLcellline,inordertoevaluatewhetherALAwouldinteracttheantitumoraleffectofbusulfan.Method: CCL-243 (K562) human cells were cultured at 37°C 5% CO2 with RPMI. Then ALA alone (0.05-1miliM) and Busulfan (20-200microM) were applied. After that optimized dosage for ALA and busulfan isdeterminedtheywereappliedasacombination in theseconcentrations (0,5mMALAve40μMBusulfan).After 24 hours, cell viability, necrosis and apoptosis wasmeasured by using Annexin-V-PI and differencesbetweengroupswereanalyzedbyMann-Whithney-UtestwithSPSS15.0program.Results:BusulfanandALAdecreasedviabilityofcellsinadosedependentmanner(p<0.05).CombinationofALA and Busulfan did not change the viability of cells compared with busulfan. Conclusion: In this study;singleALAapplicationtotumorcells,didnotproliferativeCMLcells.Besidesitshowedcytotoxiceffect.Whenit was combined with busulfan, it did not affect the cytotoxic effect of busulfan. Our next step will beevaluatingsideeffectprotectivecapacityofALAagainstbusulfantestisdamageinanimalmodelsKeywords:Chronicmyeloidleukemia,Busulfan,Alphalipoicacid


4-7

May2

016

PP-44

DIFFERENTIALEXPRESSIONSOFSIRTUINFAMILYOFGENESINBREASTCANCER

Mehriİğci²,MehmetEminKalender³,ErsinBorazan⁴ ,İbrahimBozgeyik¹,RecepBayraktar²,EsraBozgeyik²,CelaletdinCamcı³,AhmetArslan²

¹|AdiyamanUniversity,FacultyofMedicine,DepartmentofMedicalBiology,Adiyaman,Turkey²|GaziantepUniversity,FacultyofMedicine,DepartmentofMedicalBiology,Gaziantep,Turkey

³|GaziantepUniversity,FacultyofMedicine,DepartmentofOncology,Gaziantep,Turkey⁴|GaziantepUniversity,FacultyofMedicine,DepartmentofGeneralSurgery,Gaziantep,Turkey

Object:Mammaliansirtuinshavebeenshowntoperformdistinctcellularfunctionsandplaycrucialrolesinavarietyof cellularprocesses including regulationof celldeath.Also,deregulatedexpressionof thesegeneswasreportedtobe involved indevelopmentofvariousmalignancies includingbreastcancer.An increasingnumber of evidence indicates that sirtuins have both tumor promoter and tumor suppressor functions.However,therolesofsirtuinshavenotbeenwell-reportedinbreastcancer.Accordingly,inthepresentwork,weaimedtorevealtheroleofSirtuinfamilyofgenesinbreastcancer.Material and Method: In the present study, quantitative expression levels of sirtuins (SIRT1-7) in breastcancerpatientsandbreastcancercelllines(MCF-7andSKBR3)andcontrolcellline(CRL-4010)wereassessedbyusingahigh-throughputreal-timePCRmethod.Results:Asaresult,sirtuinswasfoundtobedifferentiallyexpressedinbreastcancertissuesandcancercelllines. Particularly, expressions of SIRT1 and SIRT4were found to be significantly down-regulated in breastcancertissuesandSKBR3breastcancercells.Incontrast,SIRT2,SIRT3,andSIRT5geneswereshowntobeup-regulated in our study. Although SIRT6 and SIRT7 were also up-regulated in breast cancer tissues, theseexpression changes were statistically insignificant. Additionally, SIRT2, SIRT3, SIRT5, SIRT6 and SIRT7werefoundtobedifferentiallyexpressed inbreastcancercell lines.Yet, thesechangeswerenotwell-correlatedwithtissueexpressionlevels.Conclusion:Inconclusion,sirtuinsfamilyofgenesshowsdifferentialexpressionsinbreastcancertissuesandcellsandSIRT1andSIRT4seemstoplaykeytumorsuppressorrolesinbreastcancerdevelopment.Keywords:BreastCancer;CancerCells;Expression;Sirtuins;SIRT;SIRT1


4-7

May2

016

PP-45

PAPERBASEDMOLECULARDIAGNOSISOFHUMANPAPILLOMAVIRUSATTHEPOC

MelikeKarakaya¹,²

¹|BiomedicalEngineering,BostonUniversity,Boston,MA,02215,USA,2|EngineeringSciences,IzmirKatipCelebiUniversity,Izmir,35620,Turkey

Object: Cervical cancer is a major problem for the developing world and low-resource settings wherestandard screening techniques are not accessible. The current detection methods require complex andexpensiveequipmentwhicharenotaccessibleinlow-resourcesettings.Earlydetectioniscriticalforcervicalcancer, as it is oneof the fewcancers that canbe successfully treatedwhen caughtearly.Wepropose tocreateapoint-of-care(POC)papermicrofluidicchipthatprovidesisolation,non-enzymaticamplificationanddetectionofhumanpapillomavirus(HPV),theetiologicalagentofcervicalcancer.MaterialandMethod:First,cervicalcellslysed,releasingHPVDNAthatwasextractedviapapermatrices.Thechipdoesnotrequireenzymaticamplification;electrochemicaldetectionwasusedinsteadofthetraditionalpolymerasechain reaction (PCR) that requiressilvernanoparticlesandmagneticmicrobeads.Lastly, theAgmodified DNA were detected with a hallow channel included paper device, generating electrochemicalreadout.Theentireprocesshasbeencompletedinlessthan30minutes,allowingdoctorstodiagnose,adviceandpotentiallytreatpatientsinthesamevisit.ResultandConclusion:InConclusion,Ihavereportedanewpaperextractionsupportmaterial,anovelpaperbasedelectrochemicaldetectiondevicewhich includesmodificationofDNA forPOCapplicationsofHPV16and18.IalsoappliedthemethodtothePatientsampleswhichareprovidedfromBethIsraelHospital.Itcanextractatleast10copiesperµLDNAfrompatientsamplesapproximatelyin10minutesandcandetectlowlabelconcentrationsaslowas530FMinonly4.6min.Thisdeviceis inexpensive,compatibletosimpleandrapidfabricationtechniques,userfriendly,sensitive,quantitativeandrobust.AsymptomaticpatientspositiveforHPV16and18couldbescreenedmoreclosely,thusallocatingpreciousresourcestothosemostatrisk,abeneficialuseinbothlowresourcessettingsandintheworldwideKeywords:HumanPapillomavirus(HPV),Paperfluidic,AnalyticalDetection


4-7

May2

016

PP-46

CYTOTOXICITYOFALOEVERALEAFLECTIN(ALOCTIN)COMBINEDWITHIMATINIBONAGSHUMANGASTRICTUMORCELLLINEANDSAOS-2HUMANOSTEOSARCOMACELL

LINES

EdaCandöken¹,NuriyeAkev¹,NurtenÖzsoy¹,MedihaSüleymanoğlu²,SerapErdemKuruca³

¹|DepartmentofBiochemistry,FacultyofPharmacy,IstanbulUniversity,Istanbul,TURKEY.²|DepartmentofMolecularMedicine,InstituteofExperimentalMedicine,IstanbulFacultyofMedicine,

IstanbulUniversity,Istanbul,TURKEY.³|DepartmentofPhysiology,IstanbulFacultyofMedicine,IstanbulUniversity,Istanbul,TURKEY

Object: ThebiologicaleffectsofA. veraaredue tovarious chemical compounds includinganthraquinones,glycoproteins,polysaccharides, vitaminsandenzymes. Ithasbeen reported that the lectinofA. veraplaysroleinmanybiologicalactivitiesofA.vera.Furthermore,theanticancerpotentialofAloeisalsoavailableonmanypublications.Although,severalinvitro,invivoassays,andclinicalstudieswereundertakenonA.vera,themechanismswhich accompanies the pharmacological effects of the plant have not been clarified. ThepresentstudywasdesignedtoevaluatepossiblechemopreventiveeffectsoftherecentlypurifiedAloctinanditscombinationwithimatinibmesylate(IM)invitro.MaterialandMethod:ThegelportionofleavesofA.vera(L.)Burm.f.wasseparatedfromtheleafskinanddiscarded,a single lectin (namedAloctin)was isolated from leaf skinbyammoniumsulphateprecipitation,affinitychromatography.ThecellproliferationwasdetectedbyMTTassay.AGShumangastrictumorcelllineandSaos-2humanosteosarcomacelllinewereusedtoevaluatetheeffectsofIM(25µM),Aloctin(0.5µg/ml)and their combination on cell proliferation. The concentrations of Aloctin (0,3 µg/ml – 3 µg/ml) and IM(0,1µM – 100 µM) at which concentration causing 50% inhibition of the cell proliferation (IC50) werecalculated.Results:TheresultsindicatedsignificantcytotoxicactivityofAloctinalonewithanIC50oflessthan0.3µg/ml,and of IM alone with an IC50 of 65 µM and 108 µM for AGS and Saos-2 cells respectively, while thecombinationgroupinducedhighestdecreaseincellproliferation.Conclusions:Inthisstudy,weshowedthatthecombinationofAloctinwithIMwasabletoenhancecytotoxiceffect invitro.OurresultssuggestthattheAloctincouldbeconsideredasachemotherapeuticagentwhichmay help to reveal important findings for the future use of Aloctin in cancer research by the drugcombination.

Keywords:Aloeveralectin(Aloctin),Imatinibmesylate,Cytotoxicity,AGShumangastrictumorcelllines,Saos-2humanosteosarcomacelllines.


4-7

May2

016

PP-47

INVITROEVALUATIONOFHPVSTATUSANDITSRELATIONWITHTREATMENTSTRATEGIESINHEADANDNECKCANCERS

ErsoyDoğan²,PınarErçetin¹,BarbarosAydın³,AhmetÖmerİkiz²,ZekiyeAltun¹,İlhanÖztop4,

FadimeAkman³,SafiyeAktaş¹

¹|DokuzEylulUniversitySchoolofMedicineDepartmentofBasicOncology²|DokuzEylulUniversitySchoolofMedicineDepartmentofOtorhinolaryngologyHeadandNeckSurgery

³|DokuzEylulUniversitySchoolofMedicineDepartmentofRadiationOncology4|DokuzEylulUniversitySchoolofMedicineDepartmentofClinicalOncology

Purpose:Tocompare the treatment responsesofprimarycell cultureHPVpositiveandHPVnegativeheadand neck cancers to radiotheraphy and chemotheraphy and to determine the optimal treatmentmodalitydependingontheHPVstatusofthetumor.

Method: Tumor resection sampleswere obtained from34 patientswith head and neck cancer. Single cellsuspensionswere acquired from tumor samples and cultivated in RPMI 1640medium supplementedwith10%FBS,%1penicillin/streptomycin,1%L-glutamineand1%fungisomel.8subgroupswereplannedforeachpatient’sprimarycellcultureascontrol,cisplatin,cetuximab,combinationofcisplatin+cetuximabandtheir15Gyradiotherapycombinations.Cellproliferation(BrdU),DNAdamage(H2AX)andapoptosis(PARP)levelswereanalyzedafter24hoursbyflowcytometry.ExpressionofHPV16and18wasalsodetectedbyRealtimePCR.Datawasstatisticallyanalyzedconsideringclinicaloutcomesofpatients.

Results:Therewere24maleand10femalepatientswithameanageof58.44(23-80).Tumorlocalisationwasoralcavityin10patients,oropharynxin4patients,larynxin14patientsandhypopharynxinsixpatients.HPV16waspositive in5cases(14.7%). Invitroevaluationofcytotoxic,apoptoticandDNAdamageamongHPV16 positive and negative cases did not show statistically significant difference in cisplatin, cetuximab,combination of cisplatin and cetuximab and their 15 Gy radiotherapy combinations (p:>0.05) . There wasstatistically significant difference for cell proliferetion index, apoptosis andDNAdamaged cell percentagesamongHPV16positiveandnegativecases in 15Gyradiotherapyalonegroup.Radiotherapyalonecausedmorecelldeath,apoptosisandDNAdamageintumorcellsofHPV16negativegroup.

Conclusion:Inthisinvitrostudy,chemotherapyandradiotherapysensitivityofprimarycancercellcultureofheadandneckcancerswereanalysedandcomparedamongHPVpositiveandnegativecasesbyproliferation(BrdU),DNAdamage(H2AX)andapoptosis(PARP)methods.Ourresultsindicatethatcisplatinandcetuximabdidnot have an additional effect in both groups and radiotherapywas found tobemore effective inHPVnegativecases.

Keywords:Headandneckcancer,HPV,treatmentstrategies


4-7

May2

016

PP-48

EFFECTSOFTESTOSTERONEPROPIONATEONBONEMARROWDERIVEDMESENCHYMALSTEMCELLS’PROLIFERATIONANDVIABILITY

BaşakAru¹'²,FatmaTubaAkdeniz¹'²,HüsniyeDağdeviren²'³,GülderenYanıkkayaDemirel²'³

¹|MolecularMedicineDepartment,InstituteofHealthSciences,YeditepeUniversity,Istanbul

²|ImmunologyDepartment,YeditepeUniversitySchoolofMedicine,Istanbul³|StemCellLaboratory,YeditepeUniversityHospital,Istanbul

Object:Mesenchymalstemcells(MSCs)canbefoundinvarioustissuessuchasadiposetissue,bonemarrowand umblical cord; they have CD73, CD90 and CD105 surface markers whereas they lack hematopoieticmarkers;theyareabletoattachplasticsurfaces;theycandifferentiateandhaveselfrenewalcapacity.Therearevariousstudies,indicatingeffectsoftestosteroneondifferentcelltypes(bothhealthyandcancercells).But currently effects of testosteroneonmesenchymal stem cells are unknown. In this study,we aimed toshow effects of testosterone on human bonemarrow derivedmesenchymal stem cells after 24 hours ofincubation.MaterialandMethods:WeexpandedcommerciallyavailableMSCsinvitroandaddedashort-actingformoftestosterone;testosteronepropionate(TP)atspecificconcentrations.WeusedAnnexinV–PropidiumIodideviabilityandapoptosistestfordetectingviabilityandCFSEstainingfordetectingproliferation.Measurementswereperformedonflowcytometry.Results: This study shows that testosteronepromotesproliferation in adosedependentmanner.Whereas10-8 M TP showed highest proliferation promotion and highest increase at viability. Doses over thisconcentrationwerefoundtobetoxicforMSCs.Conclusion: Our study is the first to show effects of testosterone on MSCs' viability and proliferation.TestosteroneasaculturemediasupplementmayhelptoshortenthecultureperiodforMSCs.Ourfindingsmayalsohelptofuturetreatmentstudiesofdiseaseswhicharethoughttobelinkedwithmesenchymalstemcells.Keywords:MesenchymalStemCells,TestosteronePropionate,CellCulture,Proliferation,CellDeath


4-7

May2

016

PP-49

THEEFFECTOFBONEMARROWMESENCHYMALSTEMCELLAPPLICATIONONCASPASE3ENZYMEINRATSWITHCRUSHEDMUSCLEDAMAGE.

RukiyeDemir¹,EmineDıraman¹,ErdalKaraöz²

¹|OndokuzMayısÜniversitesi,

²|CenterforRegenerativeMedicineandStemCellResearchandManufacturing(LivMedCell)

Aim:Inthisstudy,theeffectofmesenchymalstemcellapplicationonCaspase3Enzymeinratswithcrushedmuscledamagewasresearched.MaterialandMethods:Inthisstudy,therightsoleusmusclesof56femaleWistarAlbinoratswereused.Theanimalswere randomly divided as two experimental groups (aMuscleDamageGroup (MD) and aMuscleDamage+MesenchymalStemCellGroup(MD+MSC))andacontrolgroup(C).Theexperimentalgroupsweredividedintothesub-groupsofthe7th,14thand21stday(eachsub-groupn=8).Thegroupfromwhichtissuesamples were obtained without generating muscle damage was used as the control group (n=8). Tissuesamples were taken from each experimental group on the 7th, 14th and 21st day and their Caspase-3activitiesweremeasured.Thetissuesampleswereexaminedbymeansofimmunohistochemicaldyeing.Results: When the Caspase 3 activities values of the groups were compared with the control group, theincrease in theCaspase-3activities in theMD+MSCon the14thdayand21stdayswere foundstatisticallysignificant.TherewasastatisticallysignificantincreaseintheCaspase-3activitiesoftheMDonthe21stdaywhen compared toMD+MSCon the 21st day. In all theMD+MSC sub-groups, the existence ofMSCswithgreenfluorescentprotein(GFP)andthattheytransformedintomusclecellswereobserved.Conclusions:Inaddition,thefactthattheCaspase-3activityincreasedintheMSCgrouponthe14thand21stdayscanbeinterpretedasthefactthatthestemcellsreplacedthecrushedcellsandtransformedintomusclecells;andthatthecrushedcellsturnedintoapoptosis. Keywords:StemCell,Caspase-3,CrushedMuscleDamage


4-7

May2

016

PP-50

EFFECTOFTGF-Β1OVEREXPRESSIONONBIOLOGICALCHARACTERISTICSOFHUMANDENTALPULPDERIVEDMESENCHYMALSTEMCELLS

HasanSalkın¹'²,ZeynepBurçinGönen¹,ErgülErgen¹,DilekBahar¹

¹|ErciyesUniversityBetül-ZiyaErenGenomeandStemCellCenter(GENKÖK),Kayseri,Turkey

²|DepartmentofHistology-Embryology,FacultyofMedicine,ErciyesUniversity,Kayseri,Turkey

Object: In our studywere investigated how the effected the surfacemarkers,multilineage differentiation,viability, apoptosis, cell cyle, DNA damage and senesence of TGF-β1 gene therapy on human dental pulpderivedmesenchymalstromalcells(hDPSC).MaterialandMethods:hDPSCswere isolated fromhumanteethandculturedwith%20FBS inα-MEM.TheplasmidTGF-β1(SinoBiologicalInc,China)wasamplifiedinEscherichiacolihoststrainDH5αandpurifiedbyplasmid isolationwith theEndo-FreeMaxiprepPlasmid IsolationKit. TGF-β1gene transferwasperformedinto hDPSCs by electroporation method after the plasmid was prepared. For transfection efficiency wereperformed western blot and flow cytometry analyses and GFP transfection. MSC markers, multilineagedifferentiation,cellproliferation,apoptosis,cellcycle,DNAdamageandcellularsenescencewereperformedto compared transfected and non-transfected cells. Statistical analyses were performed using GraphPadPrismversion6.00forwindows(GraphPadSoftware,SanDiegoCaliforniaUSA).Results:FlowcytometryanalysisdetectedstrongexpressionofTGF-β1inpCMV-TGF-β1-transfectedhDPSCs.TGF-β1transfectionefficiencywasmeasuredas95%(Fig1D).Also,westernblotanalysisshownthatTGF-β1proteinlevelsincreasedatthirdandsixthdaysinpCMV-TGF-β1-transfectedhDPSCs.ThecontinuousTGF-β1overexpressioninhDPSCsdidnot influencetheimmunophenotypeandsurfacemarkerexpressionofMSCs.Our results showed that increased osteogenic and chondrogenic differentiation of TGF-β1 but reduced toadipogenic differentiation. Overexpression of TGF-β1 was increased proliferation and decreased totalapoptosis in hDPSCs (p<0,05). TGFB1 transfectionwas increased the number of cells at ‘S’phase (p<0,05).There was no significant difference at DNA damage. Cellular senescence decreased in TGFB1 transfectedgroup(p<0,05).Conclusion:Theseresults reflect thatTGF-β1hasmajor impactonMSCdifferentiation.TGF-β1transfectionhas no effect on cell surface markers but differentiation. TGF-β1 transfection has positive effect onproliferation,cellcycleandpreventscellularsenescenceandapoptosis.Keywords:DentalpulpderivedMesenchymalStemCells,TGF-β1Transfection,Apoptosis,DNADamage,CellularSenescence


4-7

May2

016

PP-51

MONOSODIUMGLUTAMATEHASNOCYTOTOXICEFFECTONMOUSEMESENCHYMALSTEMCELLS

SinemDal¹,SümeyyeArslan²,NejatKaanNurol³,TutkuGöktepe³,ZehraDilşadÇoban⁴,

ErtanAltaylı⁴,ŞefikGüran⁴

¹|ErzurumTechnicalUniversity,DepartmentofMolecularBiologyandGenetics²|GümüşhaneUniversity,DepartmentofGeneticsandBioengineering

³|BartınÜniversity,DepartmentofMolecularBiologyandGenetics⁴|GülhaneMilitaryMedicalAcademy,DepartmentofMedicalBiology

Aim/Backround:Monosodiumglutamateasafoodadditiveisasubstanceaddedtofoodtopreserveflavororenhance its taste and appearance.Monosodiumglutamate is the sodium salt of glutamic acid, one of themost abundant naturally-occurringnon-essential amino acids.U.S. Food andDrugAdministration reportedthatmonosodiumglutamate issafe foruse in food.However, theexcessiveuseofmonosodiumglutamatecausesdizziness,nauseaandvomiting.Duetoreports,itcausesobesity.Weaimedtofindoutthecytotoxiciyofmonosodiumglutamateandtheroleontheselectedgenes,whichhaveroleonobesity.Methods:Inourstudy,weusedmicewhichareknowntobethemostsensitiveintermsoftoxicity.Wewerestudiedthecytotoxiceffectsofmonosodiumglutamateonmousemesenchymalstemcellsatmonosodiumglutamate doses below. Also we analyzed leptin-lep and ghrelin / obestatin in prepropeptide-GHRL geneexpressionsinordertofindouttheroleofmonosodiumglutamateinobesity.Result: Monosodium glutamate below the toxic dose does not have a cytotoxic effect on mousemesenchymal stem cells. Also no expression change observed in geneswhich are known to be associatedwithobesity.Conclusion: Our results support that monosodium glutamate has no toxic effect on stem cells in uses incertain doses.Keywords: Monosodium glutamate, mouse mesenchymal stem cells, leptin gene, ghrelin/obestatinprepropeptidegene.


4-7

May2

016

PP-52

ALTEREDEXPRESSIONOFAPOPTOSIS-RELATEDGENESINCANCERSTEMCELLSINAPROSTATECANCERMODEL

HadiRouhrazi¹,NevbharTurgan¹,ÜmmüGüven²,AyşegülUysal³,GülperiÖktem³

¹|EgeUniversityInstituteofHealthSciences,DepartmentofMedicalBiochemistry,Izmir,Turkey

²|EgeUniversityInstituteofHealthSciences,DepartmentofStemCell,Izmir,Turkey³|EgeUniversityFacultyofMedicine,DepartmentofHistologyandEmbryology,Izmir,Turkey

Objective:Incontrasttolow-tumorigenicbulktumorcells(non-CSCs),cancerstemcells(CSCs)areasubsetoftumorcellswiththepotentialtoself-renewanddifferentiateintodifferentcancersubtypes.CSCsarethoughtto be responsible for tumor recurrence, distant metastasis, angiogenesis, and drug/radiation resistance.UnderstandingthepropertiesandcharacteristicsofCSCs iskeytofuturestudyoncancerresearch,suchastheisolationandidentificationofCSCs,cancerdiagnosis,andcancertherapy.Thepresentstudyfocusesonthedifferencesofexpression levelsofapoptosis-relatedgenesbetweenCSCsandnon-CSCs,usingprostateCSCsasamodelsystem.

MaterialandMethod:Clusterofdifferentiation(CD)133+high/CD44+highprostateCSCswereisolatedfromtheDU145humanprostatecancercelllinebyflowcytometrycellsorting.TotalRNAfrombothCSCsandnon-CSCswas isolatedandextracted followedby cDNAsynthesis. TheqRT-PCRarraywasused tomeasure theexpressionlevelsofapoptosis-relatedgenes.

Results:Significantup-ordown-regulationingeneexpressionwasdetectedin18genesintheprostateCSCscomparedwithnon-CSCs.ThestudiedgenescouldbeclassifiedasCaspase,BCL-2,IAP,CARD,TNFRfamiliesand genes involved in P53 signaling, NF-Kb signaling and FAS signaling pathways. By comparing the geneexpression levels between the two groups,more significant differences weremainly detected in the FAS,BIRC3, BCL2L11, FADD, TRAF5,HSP90B,HMGB1and FAM96Agenes. In addition, itwas found that severalanti-apoptoticgenesincludingBIRC3andFASweregreatlyup-regulatedinCSCs,revealinganassociationwiththe high survival rate of these cells. Conclusion: There are some differences in the expression profile ofapoptosisrelatedgenesinCSCscomparedtonon-CSCs.Thesefindingsmayprovidekeyinformationformoreeffectivetherapeuticapproaches.

Keywords:Cancerstemcell;Apoptosis;Geneexpression;Prostatecancer


4-7

May2

016

PP-53

THEPOTENTIALEFFECTSOFPROPOLISONAPOPTOSISANDCELLCYCLEPATHWAYSINHER2-POSITIVEBREASTCANCERCELLS

ÖzlemTimirciKahraman¹,MehmetFatihSeyhan¹,NeslihanSaygılı¹,HalilİbrahimKısakesen²,

AllisonPinarEronat¹,HülyaYılmazAydoğan¹,SemaBilgic3,OğuzÖztürk¹

¹|IstanbulUniversity,TheInstituteofExperimentalMedicine,DepartmentofMolecularMedicine²|IstanbulTechnicalUniversity,DepartmentofMolecularBiologyandGenetics

3|IstanbulUniversity,TheInstituteofExperimentalMedicine,DepartmentofImmunology

Object:Inthisstudy,weaimtoinvestigatetheeffectsofpropolisonER/PR-,HER2/neu+humanbreastcancercelllineSKBR3withintendtoclarifytheefficiencyofpropolisinHER2+breastcancersinoverviewofwholegenomeexpressionforthefirsttime.Material And Method: Eight different samples were collected from different regions, including China,ArgentinaandTurkey.Propolis samplesweredissolved inethanolandwater solutions.TheSKBR3 (HER2+breastcancercell)andMCF10A(normalbreastepithelialcell)wereincubatedfor24,48and72hoursafterthe addition of different concentrations of propolis extracts. Cell growth and cytotoxicity levels weremeasuredbyWST-1assay.ApoptoticcelldeathratesweredeterminedbyflowcytometryusingAnnexinV-FITC. Microarray analysis was performed to examine the gene expression profiles by using Gene-Springsoftware.Results:PropolisextractfromTurkey,50μg/mland48thhourdeterminedasthemosteffectiveextract,doseand time conditionswhichwere selected according to decrease in proliferation and increase in apoptoticeffectsofSKBR3cancercells.Atthispointforthewholegenome,wedeterminedmorerestrictedcutoff(p<0.01), totally 248 genes were significantly regulated, among which 91 were up-regulated and 157 down-regulatedinSKBR3cellsatthe48thhour.Wedetectedthatalteredexpressionlevelsof7genesoncellcyclepathwayand3genesonapoptosispathwayhada statistically significantdifference. Furthermore,no suchalterationwasdetectedinMCF10Acelllineunderthesameconditions.Conclusion:WeexhibitedthatHER2-positivebreastcancercellsarereducedproliferationchangingexpressionlevelsofcellcyclemarkersandincreasedapoptosiswithalteredexpressionlevelsofapoptoticandantiapoptoticmarkersbyİstanbul-Kartalpropolis.Thesedataprovideapotentialmechanisticbasisforoneoftheoldestnaturalagentsusedinmedicineandcancertreatment.Keywords:Propolis,HER2-PositiveBreastCancer,Apoptosis,CellCycle,Microarray


4-7

May2

016

PP-54

FERULICACIDDECREASESCELLVIABILITY,INVASION,MIGRATIONANDCOLONYFORMATIONINMIAPACA-2HUMANPANCREATICCANCERCELLSINVITRO

UmutFahrioğlu² ,YavuzDodurga¹,LeventElmas¹ ,MücahitSeçme¹

¹|PamukkaleUniversity,FacultyofMedicine,DepartmentofMedicalBiology,Denizli,Turkey

²|NearEastUniversity,FacultyofMedicine,DepartmentofMedicalBiology,Nicosia,NorthCyprus

Objects:Novelandcombinatorialtreatmentmethodsarebecomingsoughtafterentitiesincancertreatmentandthesetreatmentsareevenmorevaluableforpancreaticcancer.Thescientistsarealwaysonthelookoutfornewchemicalstohelpthemintheirfightagainstcancer.Inthisstudy,weexaminetheeffectsofferulicacid (FA),aphenolic compound, found invariousnutrients,ongeneexpression,viability, colony formationandmigration/invasionintheculturedMIAPaCa-2humanpancreaticcancercell.Material andMethod: Theeffect of FAon cell viabilitywasdeterminedbyCellTiter-Glo® LuminescentCellViabilityAssay.ExpressionprofilesofcertaincellcycleandapoptosisgenessuchasCCND1,CDK4,CDK6,RB,p21,p16,p53,caspase-3,caspase-9,caspase-8,caspase-10,Bcl-2,BCL-XL,BID,DR4,DR5,FADD,TRADD,PARP,APAF, Bax, Akt, PTEN, PUMA,NOXA,MMP2,MMP9, TIMP1 and TIMP2were determinedby real-time PCR.Furthermore, effects of FA in MIA PaCa-2 cells on invasion, colony formation and cell migration weredeterminedbymatrigelchamber,wound-healingtestandcolonyformationassay,respectively.Results:IC50doseinMIAPaCa-2cellswasdetectedas500μM/mlatthe72ndhour.Additionally,effectsofFA on colony formation and invasionwere also investigated. It was observed that FA caused a significantdecrease in the expression of CCND1, CDK4/6, Bcl2 and caspase 8 and 10 in the MIA PaCa-2 cells whilecausinganincreaseintheexpressionofp53,Bax,PTENcaspase-3and9.FAwasobservedtodecreasecolonyformation,invasionandmigrationofMIAPaCa-2pancreaticcancercells.Conclusion:FA is thought tobehaveasananti-canceragentbyaffectingcell viability, cell cycle,apoptotic,invasionandcolonyformationbehaviourofMIAPaCa-2cells.Therefore,FA isplacedasastrongcandidateforfurtherstudiesaimedatfindingabetter,moreeffectivetreatmentapproachforpancreaticcancer Keywords:Ferulicacid,Pancreaticcancer,Apoptoticgenes,Cellcyclegenes


4-7

May2

016

PP-55

ANALYSINGTHECYTOTOXICEFFECTSOFCERANIB-2ONHEPATOCELLULARCARCINOMACELLS

DjananVejselova¹,HatIceMehtapKutlu¹

¹|DepartmentofBIology,FacultyofScIence,AnadoluUnIversIty,YunusemreCampus,26470,EskIşehIr,

Turkey

Object: CeramIde Induces cell death In cancer cells on the contrary sphIngosIne-1-phosphate leads to cellprolIferatIon.CeramIde/sphIngosIne-1-phosphateratIoIscrItIcalforthevIabIlItyofthecellsandthIslevelIsregulated by ceramIdase enzymes. CeranIb-2, an antIcancer agent Is an InhIbItor of human ceramIdases.InhIbItIngofceramIdasescauseImbalanceInceramIde/sphIngosIne-1-phosphateratIo.HereIn,ceranIb-2wasInvestIgatedforItscytotoxIceffectsInhumanhepatocarcInomacellsHep3B2.MaterIal and Method: Stock solutIon of ceranIb-2 was prepared In dImethyl sulphoxIde (DMSO; SIgmaAldrIch,UK)andfurtherdIlutIonsweremadewIthfreshculturemedIumIntherangeof5-110µM.MTT(3-(4,5-dImethylthIazol-2-yl)-2,5-dIphenyl-2H-tetrazolIumbromIde)colorImetrIcassaywasusedtofIndthecellvIabIlItyfor24hours.IC50valuewasdetermInedaccordIngtotheELISAreader(ELx808)results.Results: The vIabIlIty of Hep3B2 cells reduced In dose-dependentmanner. In our results It Is shown thatceranIb-2exhIbItedhIghantIprolIferatIveeffectsonHep3B2cellsbeIngcytotoxIcInlowconcentratIons.ConclusIon: On the basIs of our fIndIngs ceranIb-2 Is found to be a good InhIbItör of cell prolIferatIon ofHep3B2cellsandwesuggestthIsagentforfurtherInvestIgatIonfordrugdesIgnIngforcancertherapy. Keywords:HumanhepatocarcInoma,LIvercancer,CytotoxIcIty


4-7

May2

016

PP-56

CYTOTOXICEFFECTOFMORUSRUBRAEXTRACTONHUMANCOLONCANCERCELLSTHROUGHINCREASEDENDOPLASMICRETICULUMSTRESSANDSUPPRESSED

TELOMERASE

SelimDemir1,İbrahimTuran2,3,KağanKılınç2,SemaMısır4,5,SerapÖzerYaman4,ElifAyazoğluDemir6,AyşeArslan7,AhmetMenteşe4,YükselAliyazıcıoğlu4,OrhanDeğer4

¹|DepartmentofNutritionandDietetics,FacultyofHealthSciences,KaradenizTechnicalUniversity,61080,

Trabzon,²|DepartmentofGeneticandBioengineering,FacultyofEngineeringandNaturalSciences,Gumushane

University,29100,Gumushane,³|MedicinalPlants,TraditionalMedicinePracticeandResearchCenter,GumushaneUniversity,29100,

Gumushane,⁴|DepartmentofMedicalBiochemistry,FacultyofMedicine,KaradenizTechnicalUniversity,61080,Trabzon,

⁵|DepartmentofBiochemistry,FacultyofPharmacy,CumhuriyetUniversity,58140,Sivas,6|DepartmentofChemistry,FacultyofSciences,KaradenizTechnicalUniversity,61080,Trabzon,

7|DepartmentofBiotechnology,InstituteofScience,GumushaneUniversity,29100,Gumushane,Turkey.

ABSTRACTObject:Cancercellshavesomecharacteristicfeatures,suchasevasionofapoptosis,dysregulationofcellcyclecontrol,self-sufficiencyingrowthsignalling,andtelomeraseactivation.Telomerasethereforehasanimportantroleinunlimitedproliferationofcancercells,soinhibitionoftelomeraseactivityisapotentiallyhighlyselectivetargetforcancertreatment.Inrecentyears,inductionofendoplasmicreticulum(ER)stress-mediated apoptosis by CCAAT/enhancer-binding protein homologous protein (CHOP) in cancer cells isrepresentedasanalternativeapproachforcancertherapy.Morusrubra,knownas"redmulberry",belongstofamilyofMoraceaeandgenusofMorus.Itisrichinpolyphenolsanditsbiologicalfunctionsareattributedtothesecompounds.ManystudieshaveevaluatedthecytotoxiceffectsofdifferentMorusspecies,butthereisnostudyaboutcytotoxiceffectofM.rubraoncoloncancercells.TheaimofthisstudywastoinvestigatethecytotoxiceffectoftheM.rubraextractoncolon(WiDr)cancercellsintermsofERstressandtelomeraseexpressionforthefirsttime.MaterialandMethod:ThecytotoxicactivityofextractwasdeterminedbyMTTassay.TheeffectofM.rubraextracton telomeraseandCHOPexpressionwasdemonstratedbyRT-PCRtechnique.Results:Morus rubraextract exhibited selective cytotoxicity on colon cancer cells compared tonormal fibroblast cells.M. rubraextractsignificantly repressedthe telomeraseexpressionatall treatment time (24,48,and72h) inadosedependentmanner in colon cancer cells.Moreover, treatmentwithM. rubra extract significantly inducedCHOPexpressionat72hinthesecells.Conclusion:OurresultsdemonstratedthatM.rubraextractinducescytotoxicityincoloncancercellsthrough,atleastinpart,increasingERstress-relatedsignallingandrepressingtelomeraseexpression.


4-7

May2

016

PP-57

MORUSRUBRAEXTRACTINDUCESAPOPTOSISANDINHIBITSPROLIFERATIONINHUMANPROSTATECANCERCELLS

İbrahimTuran1,2,SelimDemir3,KağanKılınç1,YükselAliyazıcıoğlu4,AhmetAlver4,SemaMısır4,5,

SerapÖzerYaman4,KübraAkbulut4,OrhanDeğer4

¹|Department of Genetic and Bioengineering, Faculty of Engineering and Natural Sciences, Gumushane

University,29100,Gumushane,

²|Medicinal Plants, Traditional Medicine Practice and Research Center, Gumushane University, 29100,

Gumushane,

³|DepartmentofNutritionandDietetics,FacultyofHealthSciences,KaradenizTechnicalUniversity,61080,

Trabzon,

⁴|DepartmentofMedicalBiochemistry,FacultyofMedicine,KaradenizTechnicalUniversity,61080,Trabzon,

⁵|DepartmentofBiochemistry,FacultyofPharmacy,CumhuriyetUniversity,58140,Sivas,Turkey.

ABSTRACTObject:Cancerisaheterogeneousdisease,twoofwhosecharacteristicfeaturesareuncontrollablecell proliferation and insufficient apoptosis. Morus rubra, known as "red mulberry" belongs to family ofMoraceaeandgenusofMorus.Infolkmedicines,thefruitsofMorusspeciesareusedtotreatmanydiseasesand their biological effects are attributed to their phenolic contents. Many studies have evaluated thecytotoxiceffectsofdifferentMorusspecies,butthereisnostudyaboutcytotoxiceffectofM.rubra.Inthisstudy,weaimedtoevaluatephenoliccontent,phenoliccomposition,andcytotoxiceffectofM.rubraextract.Material and Method: Total polyphenol content, phenolic composition, and cytotoxic effect of M. rubraextract were determined using Folin-Ciocalteu method, HPLC, and MTT assay, respectively. Then,mechanismsofcytotoxiceffectofM.rubraextractonhumanprostate(PC-3)cancercellswereexaminedinregardtocellcycle,andapoptosisusingflowcytometricmethods.Results:TotalpolyphenolcontentofacidifieddimethylsulfoxideextractofM.rubrawasfoundas11.9±0.1mg GAE per to g sample. Ascorbic acid, gallic acid, and epigallocatechin gallate were detected as majorantioxidant compounds in the extract. M. rubra extract exhibited selective cytotoxicity against prostatecancer cells compared tonormal fibroblast cells.Wedetermined thatM. rubraextract increasedcell cyclearrestatG1phaseandexhibitedapoptoticfeaturesinprostatecancercells.Conclusion:Our results showed thatM. rubraextracthasapoptoticandantiproliferativeeffect inprostatecancer cells. Further studies are needed to better understand the molecular mechanisms underlying thiseffectofM.rubraextractKeywords:Apoptosis,CellCycle,Cytotoxicity,Morusrubra,ProstateNeoplasms


4-7

May2

016

PP-58

THEEFFECTSOFRESVERATROLGAINIMPORTANCEACCORDINGTOP53MUTATIONINHCT116COLONCANCERCELLLINES

GüneşÖzen¹'²,BelginSertSerdar²,HalilAteş³,SemraKoçtürk²

¹|DokuzEylülUniversity,InstituteofHealthSciences,DepartmentofMolecularMedicine,35340,Inciraltı,

Izmir²|DokuzEylülUniversity,FacultyofMedicine,DepartmentofBiochemistry,35340,Inciraltı,Izmir

³|DokuzEylülUniversity,FacultyofMedicine,DepartmentofHematology,Izmir,Turkey

Cancer is a disease that includes heterogenic and complexmolecular changes. Anti-carcinogenic effects ofResveratrol, anaturalpolyphenol,havebeenproved inavarietyof cancer cells.Considering theeffectsofResveratrol, the influenceof the signal transductionpathways in thepresenceor absenceof p53of coloncancercells isgaining importance.Ouraimwas to investigate theeffectsofResveratrol in thepresenceorabsence of p53 on cell viability, apoptotic cell death ratio and fold changes of proliferative or anti-proliferative gene expressions, which may have important effects on colon cancer, in HCT116 coloncarcinomacells.IC50dosesofResveratrolweredeterminedbyWST-1assay.Theapoptoticcelldeathratiosintreatments of ResveratrolweredeterminedbyAnnexin-V-FITC/PI assay for flow cytomety. The changes ofCCND1,FRA1,PPARD,EGFR,BIRC5,PCNA,MCL1,STAT3,FOS,JUN,P27,ATF4,TRAIL,PUMA,GADD45A,RB1,FASLG, TNF, SOCS3, STAT1 gene expressions were evaluated by Real Time PCR. All data were statisticallyanalysed by Student’s t test. Our research has revealed that Resveratrol (60μM) causes decrease in cellviability and increase in apoptotic cell death in HCT116p53(+/+) and HCT116p53(-/-) cells significantly(p<0.05).ThefoldchangesofthegeneexpressionshaveshownthatResveratrolhassignificant(p<0.05)anddifferenteffectsontheexpressionsofthegenesrelatedwiththeexistenceofp53inHCT116celllines.Hencewe proposed that Resveratrol might show proliferative or apoptotic effects related with p53mutation ofcolon cancer cells andwe predicted that unconscious consumption of Resveratrol in colon cancer patientmightcauseadverseeffects.ThisstudywassupportedwithScientificResearchProjectsCoordinationUnitofDokuzEylulUniversitywith2013.KB.SAG.055projectnumber Keywords:Resveratrol,p53,ColonCancer,SignalTransductionPathways


4-7

May2

016

PP-59

THEEFFECTSOFMIRTAZAPINEONDIABETICNEPHROPATHY

EzgiBektur¹,ErhanŞahin¹,CengizBayçu¹

¹|EskişehirOsmangaziÜniversitesiDiabetesmellitusisaserioushealthproblemandisassociatedwithsevereacuteandchroniccomplicationsthat influencequalityof life.Diabeticnephropathyhasbecamethemostcommoncauseofend-stagerenaldisease. Hyperglycemia has been shown to induce in vitro apoptosis of several cells and apoptosis inendothelialcells indiabeticvascularcomplications.Mirtazapineisanoradrenergicandspecificserotonergicantidepressant. It is also known that mirtazapine is shown to inhibit the production of enzymatic/ non-enzymaticoxidantparameters.Weaimthathowmirtazapineeffectondiabeticnephropathy.A totalof21maleSpragueDawleyrats,8weeksoldandweighting180-200g,werepurchasedfromEskisehirOsmangaziUniversity Medical and Surgical Experimental Research Center (Eskisehir, Turkey). 14 rats were randomlyselectedand injectedonce intraperitoneallywith streptozotocin (55mg/kg).After72h,blood-glucose levelsweremeasuredbyglucometerandlevelshigherthan300mg/dlwereacceptedfordiabetes.After4weeks,7outofthe14ratswasadministered20mg/kgfor14daysbyintragastricgavage.Controlgroupwasinjectedwithanequalvolumeofcitratebuffer.Incontrolgrouphealthycortexlayer,negativeCaspase1,highNLRP3andnegativeTUNELreactionswereseen.IndiabetgroupClassIIbdiabeticnephropathy,diffuseexpansionofmesangium,tubularatrophy,highexpressionofCaspase-1andNLRP3inglomerularendothelialandtubulesand TUNEL positive reaction in glomerul and tubular nucleus were seen. In Mirtazapine group healthyglomerulus, low expression of Caspase 1,mildNLRP3 expression ofNLRP3 and TUNEL positive reaction inglomerulusand tubularnucleus (Figure1).Parallel reactionswere seen inwesternblot too (Figure2).OurresultsshowthatdiabetesmellitusinducedapoptosisbutnotbyNLRP3inflamationpathwayandmirtazapineadministrationtothediabeticnephropathymaybeworthconsideringasanewcandidateforpreventingcelldeath.Keywords:Diabeticnephropathy,mirtazapine,NLRP3


4-7

May2

016

PP-60

TUBULARCELLAPOPTOSISINPATHOGENESISOFTYPE1CARDIORENALSYNDROMEANDEFFECTOFERYTHROPOIETIN

AyselGüvenBağla¹,MeltemİçkinGülen²,ErtuğrulErcan³,FatihAşgün⁴

¹|SANKOUniversity,SchoolofMedicine,DepartmentofHistologyandEmbryology,Gaziantep

²|ÇanakkaleOnsekizMartUniversity,SchoolofMedicine,DepartmentofHistologyandEmbryology,Çanakkale

³|İzmirUniversity,SchoolofMedicine,DepartmentofCardiology,MedicalParkHospital,Izmir⁴|ÇanakkaleOnsekizMartUniversity,SchoolofMedicine,DepartmentofCardiovascularSurgery,Çanakkale

Objective:Condition of renal failure occurring in heart failure is termed cardiorenal syndrome. This isattributed to hemodynamic derangement, with reduced renal perfusion and increased venous pressure.Erythropoietin(EPO) isacytokine,withsignificanthomologytomediatorsofgrowthandinflammation.WeaimedtoresearchearlychangesinrenaltissueaftermyocardialinfarctionandeffectofEPOonthistissue.

MaterialandMethods:CoronaryarteryligationwasperformedtoinducemyocardialinfarctioninWistarrats.Rats (n=36)weredivided into five groups;Group1: Sham-operated control rats.Group2:Rats receivingasingleintraperitonealinjectionofsalineimmediatelyafterligationandsacrificed6hoursaftersurgery;Group3: Rats receiving a single intraperitoneal injection EPO (5000 U kg−1) (Eprex 4000 IU/0.4 mL)immediatelyafterligationandsacrificed6hoursaftersurgery;Group4:RatsreceivingasingleintraperitonealinjectionEPO(10000Ukg−1)immediatelyafterligationandsacrificed6hoursaftersurgery;Group5:Rats without treatment sacrificed 1 hour after ligation. We investigated changes with light microscopy,immunohistochemistry(caspase3).

Results:We observed renal morphology was regular in Group 1 and Group 5. However in Group 2, weobserved tubular infiltration (p=0,007), tubular necrosis (p=0,000), tubular dilatation (p=0,015), luminalcongestion (p=0,020)with loss of brush border (p=0,000) and spillage of epithelial cells into tubule lümen(p=0,000).SignificantlylesstubulardamagewasobservedinEPO-treatedgroups(Group3andGroup4).

Caspase3 immunostainingwasobservedat6hours in renal cortex (p=0,002) (Group2),at1hour in renalmedulla (p=0,016) (Group 5). Caspase-3 immunostaining decreased inthe medulla of EPO-treated groups(p=0,016)(Group3andGroup4).

Conclusion:Cardiacreasonsthatcancauseacutekidney injury influencesparticularlythemedulla inaveryshort time.EPOtreatmentpromotes aprotective effecton thecardio-renal axis,whichmightbe attributedtoitsanti-apoptoticproperties.

Keywords:Erythropoietin,caspase3,cardiorenalsyndrome


4-7

May2

016

PP-61

ANOVELREGULATOROFMITOPHAGYISAPUTATIVEAUTOPHAGY-UPSCOORDINATORPROTEIN

NurM.Kocaturk¹,KarinEberhart¹,DevrimGözüaçık¹

¹|SabancıUniversity

Autophagyandubiquitinproteasomesystemsarethemajordegredationsytemsinmammaliancellsfortherecycling of cellular contents ranging from soluble proteins to intracellular organelles. Previous studiesdemonstrated these two independentdegredationpathwayshave compansatory effetoneachotherwhenoneofthemisinhibitedinordertopreventcellsfromtoxicityoftheproteinaggregatestheotherpathwayisactivated. Additionally, recent findings suggest that mitophagy and UPS work coordinatively in order toeliminatedysfunctionalmitochondria.HowevernodirectconnectionbetweenautophagyandUPShasbeenidentified untill now. Due to the role ofmitochondrial function in aging and age-related diseases such asParkinson’s disease, Alzheimer’s disease and Huntington’s disease, understanding the nature ofmitochondrialdynamicshasgreatimportance.ThepurposeofthestudytointroduceforthefirsttimedirectlinkbetweenautophagyandUPSintheregulationofmitochondrialhomeostasis.Protein-proteininteractionsevaluatedthroughcolocalizationstudies,immunoprecipitationandgelfiltrationtechnique.Totalcell lysateswereanalysedforinvitrodeterminationofthemitochondrialproteindegredation.Subcellularfractionationexperimentsperformed for thedeterminationof thestress-induced recruitmentof cytoplasmicproteins tomitochondria. In this study, we show that proteasome shows direct physical interaction with autophagicmachinery under mitochondrial stress conditions. The interaction between autophagic machinery andproteasomesubunitisimportantforretranslocationofcytoplasmicproteinstomitochondria.Takentogether,ourstudyintroducedaregulatorysubunitofproteasomeasanovelregulatorofmitophagyandaninteractorofATG5,anddescribedasadirectcoordinatorofmitochondrialclearanceandUPS.Thesefindingsmighthaveimplicationsforproteinopathiesinvolvingmitophagydefects,includingParkinson'sDisease.ACKNOWLEDGEMENT: THIS WORK WAS SUPPORTED BY THE SCIENTIFIC AND TECHNOLOGICAL RESEARCHCOUNCILOFTURKEY(TUBITAK)1001GRANT(PROJECTNO.114Z241)ANDN.M.K.ISRECIPIENTOFTUBITAK-2210DOCTORALSCHOLARSHIP.Keywords:Autophagy,Proteasome,UPS,MItochondrIa,MItochondrIalDynamIcs


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PP-62

AUTOPHAGYTARGETSANIBMPFD-RELATEDVCP/P97MUTANTPROTEIN

ÖznurBayraktar¹,ÖzlemOral¹,NurMehpareKocatürk¹,KarinEberhart¹,AliKoşar¹,DevrimGözüaçık¹

¹|SabanciUniversity,Turkey

Here,westudIedtheroleofanIBMPFD-relatedmutantofVCP/P97(P137Lmutant)Inautophagy.IncontrastwIthwIld-typeVCP/P97orcommonlystudIedmutants(R155orR191),theP137mutantwasaggregate-prone.Weshowedthat,unlIkeothermutants,theP137mutantproteInstImulatedbothautophagosomeandautolysosomeformatIon.Moreover,P137mutantproteInItselfwasasubstrateofautophagy.StarvatIon-andmTORInhIbItIon-InducedautophagyledtothedegradatIonoftheP137mutant,whIlepreservIngwIld-typeandfunctIonalVCP/P97.OurresultsIndIcatethatcellularmechanIsmsleadIngtoIBMPFDdIseasemaybevarIous,andunderlInetheImportanceofstudyIngdIfferentdIsease-assocIatedmutatIonstobetterunderstandhumanpathologIesandtaIlormutatIon-specIfIctreatmentstrategIes.Keywords:Autophagy, VCP, IBMPFD, autophagosomematuratIon, autophagy target, ubIquItIn-proteasomesystem,mItophagy


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PP-63

OXADIAZOLE-BASEDPYRAZOLINEDERIVATIVESASNEWANTITUMORAGENTS

AhmetÖzdemir¹,MehlikaDilekAltıntop¹,BelginSever¹,ÖzlemAtlı¹,MerveBaysal¹,ZaferAsımKaplancıklı¹

¹|AnadoluÜniversitesi

Inrecentyears, thedevelopmentofresIstancetoantIcancerdrugshasemergedasamajorobstacle InthefIghtagaInstcancerandthereforeaconsIderableamountofresearchhasbeencarrIedoutonthedIscoveryofnewselectIveantIcanceragentswIthenhancedactIvItyand lImItedtoxIcIty[1,2]. InanefforttodeveloppotentantIcanceragents,someoxadIazole-basedpyrazolInederIvatIvesweresynthesIzedvIathereactIonof1-(chloroacetyl)-3-(2-furyl)-5-aryl-2-pyrazolIneswIth5-substItuted-1,3,4-oxadIazole-2(3H)-thIones.MTTassaywascarrIedouttodetermInecytotoxIceffectsofthecompoundsonMCF-7humanbreastadenocarcInoma,A549 human lung adenocarcInoma and NIH/3T3 mouse embryonIc fIbroblast cell lInes. 1-[(5-Benzhydryl-1,3,4-oxadIazol-2-yl)thIoacetyl]-3-(2-furyl)-5-phenyl-4,5-dIhydro-1H-pyrazole was found to be the mosteffectIveagent InthIsserIesagaInstMCF-7cell lInewIthan IC50valueof125µg/mLwhencomparedwIthcIsplatIn(IC50=35.31µg/mL)andlowtoxIcItytoNIH/3T3celllIne(IC50>500µg/mL).Ontheotherhand,thIscompound showed low antIcancer actIvIty agaInst A549 cell lIne (IC50= 339 µg/mL)when comparedwIthcIsplatIn(IC50=42.57µg/mL).Keywords:OxadIazole,PyrazolIne,Cancer,DrugDesIgn


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PP-64

DNASYNTHESISINHIBITORYEFFECTSOFNEWTHIAZOLEDERIVATIVES

MehlikaDilekAltıntop¹,AhmetÖzdemIr¹,GülşenAkalınÇiftçi2

¹|DepartmentofPharmaceuticalChemistry,FacultyofPharmacy,AnadoluUniversity,26470Eskişehir,Turkey²|DepartmentofBiochemistry,FacultyofPharmacy,AnadoluUniversity,26470Eskişehir,Turkey

CancerIsoneoftheleadIngcausesofdeaththroughouttheworldandthereforeextensIveeffortshavebeendevotedtothedIscoveryofnewpotentandselectIveantIcanceragents.TheclInIcaleffIcacyoftIazofurInandItsanalogues,bleomycInsanddasatInIbpoIntedouttheImportanceofthIazolescaffoldInthefIeldofcancertreatment.1 In the currentwork, new thIazolyl hydrazone derIvatIveswere synthesIzed and evaluated fortheIrcytotoxIceffectsonA549human lungadenocarcInoma,C6ratglIomaandNIH/3T3mouseembryonIcfIbroblastcelllInesusIngMTTassay.Furthermore,CellProlIferatIonELISA,BrdU(colorImetrIc)kItwasusedtodetermIne DNA synthesIs InhIbItory effects of the most effectIve compounds. 2-[2-(4-(1H-1,2,4-trIazol-1-yl)benzylIdene)hydrazInyl]-4-(4-nItrophenyl)thIazolewas themost promIsIng agent In thIs serIes due to ItssIgnIfIcant InhIbItory effect on C6 cell lIne wIth an IC50 value of 13.00±1.00 µg/mLwhen comparedwIthcIsplatIn (IC50=12.67±3.06 µg/mL) and low cytotoxIcIty agaInst NIH/3T3 cell lIne (IC50= 733.33±256.58µg/mL).DNAsynthesIsInhIbItIonpercentofthIscompoundwas62.2%atItsIC50value(13.00±1.00µg/mL),whereastheInhIbItIonpercentofcIsplatInwas53.95%atIC50value(12.67±3.06µg/mL).Ontheotherhand,thIscompounddIdnotshowsIgnIfIcantcytotoxIcactIvItyagaInstA549celllIne(IC50=453.33±80.83µg/mL). Keywords:ThIazole,Cancer,DNASynthesIs


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PP-65

SYNTHESISANDEVALUATIONOFDNAINHIBITORYEFFECTSOFNOVELTHIOSEMICARBAZONEDERIVATIVES

BelginSever¹,MehlikaDilekAltıntop¹,GülşenAkalınÇiftçi¹,ZaferAsımKaplancıklı¹

¹|AnadoluÜniversitesi

CancerIsaverycommoncondItIonwhIchaffectspeopleallovertheworld.LotsofdIseasesoralteratIonscancausecancersoIt IsnotasIngledIsease.AchangeoccursInthebodythencellsstarttogrowandmultIplyuncontrollably1. ThIosemIcarbazones possess a wIde range of bIologIcal effects IncludIng antIneoplastIc,antIbacterIal, antIfungal and antIvIral effects and they gIve rIse to a great varIety of coordInatIonmodesdependIngontheIrchemIcalstructure2.TrIapIne,athIosemIcarbazonederIvatIve,IsapromIsIngantIcanceragent InhIbItIngrIbonucleotIdereductaseenzyme,whIch Is Important forcellprolIferatIon3. In thecurrentwork, 4-substItuted benzaldehyde N-(1,3-benzodIoxol-5-yl)thIosemIcarbazones were synthesIzed andevaluatedfortheIrcytotoxIceffectsonC6ratglIomaandNIH/3T3mouseembryonIcfIbroblast(healthy)celllInes usIng MTT assay. Cell ProlIferatIon ELISA, BrdU (colorImetrIc) kIt was also used to determIne DNAsynthesIs InhIbItory effects of the most effectIve compounds. BIphenyl substItuted compound can beIdentIfIedasthemostpromIsIngantIcanceragentagaInstC6celllIne(IC50=32.67±6.43μg/mL)comparedtocIsplatIn(IC50=14.33±2.31μg/mL).Furthermore,thIscompoundshowednosIgnIfIcantcytotoxIcItyagaInstNIH/3T3 cell lIne (IC50= 143.33±32.14). On the other hand, DNA synthesIs InhIbItIon percent of thecompoundwasfoundas54.99%,whereastheInhIbItIonpercentofcIsplatInwasfoundas53.95%.

Keywords:ThIosemIcarbazone,Cancer,DNASynthesIsInhIbItoryEffect


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PP-66

INVESTIGATIONOFCARMOFURINDUCEDULTRASTRUCTURALCHANGESONHUMANLUNGADENOCARCINOMACELLS

EmreÇömlekçi¹,DjananVejselova²,HaticeMehtapKutlu²

¹|DepartmentofMolecularBIologyandGenetIcs,GraduateSchoolofScIences,BIlecIkŞeyhEdebalI

UnIversIty,BIlecIk,Turkey²|DepartmentofBIology,FacultyofScIence,AnadoluUnIversIty,EskIşehIr,Turkey

Object: The IncIdence of cancer has been experIencIng arIse In recent years. Lung cancer Is one of typecommoncasesIntheworld.LungcancerIsoneoftypecommoncasesIntheWorld. InTurkey, lungcancerIncIdanceInmansIsatabout%62and%5Inwomans.ThefrequentcaseIncancertherapyIsdevelopIngaresIstance to antI-cancer agent. Thus, there Is a need for new antI-cancer agents. Carmofur, a prImIdIneanalogue, has been used In colorectal cancer therapy. In the present study we InvestIgated the cell theeffectsofcarmofurontheA549cellsultrastructure.MaterIalandMethod:ForultrastructuralevaluatIonsastocksolutIonofcarmofurwaspreparedIndImethylsulfoxIde(DMSO)andtheIC50concentratIondetectedInourprevIousstudywasapplIedfor24hourstotheA549cells and fIxed InglutaraldehydeovernIghtat+4 °C. FollowIng fIxatIon,A549cellswerepost-fIxed InosmIumtetroxIde,dehydratedInethanolthenembeddedInEPON812epoxyresIn.SapmlesweresectIonedand staIned In uranyl acetate and lead cItrate then observed under a transmIssIon electron mIcroscope(TEM).Results: Inour resultswedemonstrated that carmofurwashIghly cytotoxIc In lowdoses InA549cellsandcaused damages In the cell ultrastructure namely ondulatIons and blebbIngs on the cell membrane,condensatIon and fragmentatIons of the nucleI, shrInkage of the cells that IndIcate apoptosIs that Is thepreferedcelldeathIncancercelllInes.ConclusIon:Fromourresults ItcanbeconcludedthatcarmofurhasabIgpotentIal InkIllIngtheA549cells.AfterfurtherInvestIgatIonscarmofurmaybeclaImedasanusefulagentforantI-cancerdrugdesIgnIng.

Keywords:Carmofur,lungadenocarcInoma,cytotoxIcIty,TEM


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INDEXAAçıkyıldızM.,74,76AggarwalBB.,24AghaDM.,40AğcaCA.,58,103AhmadovU.,43AkanP.,89AkbabaE.,50AkbulutK.,125AkçalıS.,63AkçaH.,105AkçoraCM.,87AkdenizFT.,116AkevN.,114AkgülB.,43,62,64AkınMN.,50AkkoçY.,95AklH.,40AkmanF.,115AksoyM.,111AksunS.,92AktaşA.,93AktaşS.,66,111,115AlaşehirliB.,72,73AlcıgırME.,37AliyazıcıoğluY.,124,125AllmerJ.,43AltaylıE.,119AltıntopMD.,131,132,133AltopA.,91AltunK.,66AltunZ.,66,115AltundağEM.,45,52,60,82AlverA.,125ArısanED.,21Armağanİ.,71ArslanA.,55,112,124ArslanME.,74,76ArslanS.,119AruB.,116AşgünF.,128AşkınH.,58,103AruB.,77AtabayMN.,91AteşH.,54,57,126AtlıÖ.,131

AtmacaB.,109AvcıÇB.,59AydemirE.,96AydınB.,115AydoğanHY.,51,121BBağcıC.,43BağcıG.,105BağlaAG.,128BaharD.,118BarthND.30BaşHH.,99BatırelHF.,60BatırelS.,60BayHH.,42BayçuC.,127BayraktarÖ.,41,130BayraktarR.,112BaysalM.,131BekturE.,127BerehabM.,40BeresteinGL.,32BergquistJ.,42BerrakÖ.,21BeyzadeogluM.,81BilgiçS.,121BilirA.,47BoraU.,89BorazanE.,112BozaykutP.,45BozgeyikE.,55,107,112Bozgeyikİ.,55,107,112BronD.,40BulutF.,92BulutÖ.,83BurnyA.,40BütünerBD.,36BüdeyriS.,99CCamcıC.,53,97,112CanA.,23CanacankatanN.,39CanbolatMF.,70CandökenE.,114

CecconiF.,17CellatM.,84CengizB.,53,97CengizH.,57ChaoMW.,38ÇÇağlarÖ.,39ÇalanGÖ.,89ÇalışırM.,50ÇelebiS.,57ÇelenÇ.,75ÇetinHO.,92ÇevikMÖ.,55ÇiçekM.,61ÇiftçiGA.,132,133ÇobanZD.,109,119ÇolakAT.,106ÇömlekçiE.,134DDağdevirenH.,116DalS.,119DeğerO.,124,125DemirEA.,124DemirH.,109DemirR.,117DemirS.,124,125DemirelGY.,77,116DemiryürekAT.,53,72,73,97DemiryürekŞ.,53,72,73,97DengjelJ.,69DereticV.,14DıramanE.,117DikmenBY.,37DilmaçS.,100DirenA.,51DodurgaY.,59,122DoğanE.,115DoğanHO.,37DoğanK.,37DoğantürkZ.,73DökmeciS.,41DoranF.,39DransfieldI.,30DüzgünerV.,84,85,91


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016

EEberhartK.,129,130EkiciAID.,95EkinciA.,47EkinciC.,47ElmasL.,59,122EngelwardBP.,38EpikmenET.,78ErbaykentBT.,66ErbilS.,69ErcanE.,128ErcanG.,49,56,110ErçetinP.,111,115ErdoğanE.,81ErdoğanMK.,58,103ErgenE.,118ErinN.,100ErkekoğluP.,38EroğluC.,59EronatAP.,121ErsenDA.,89FFahrioğluU.,122GGençY.,86GeyikoğluF.,74GhanemG.,40GhanitabeN.,57GöçmenB.,75,94GökY.,93GökalpS.,63,98GökçeG.,69GöktepeT.,119GönenZB.,118GörgünC.,44,46GözüaçıkD.,15,41,69,95,129,130GruneT.,42GülaçtıF.,69GülenMİ.,128GülerS.,64GültekinF.,70GüranŞ.,109,119GümüşR.,81GümüşelBK.,38

GümüşkaptanÇ.,21GündemE.,81GündüzR.,72,73GürkanAÇ.,21GüvenÜ.,120,135GüvençY.,83HHarputÜŞ.,86HeebMJ.,30HepdumanC.,92HepokurC.,61İİğciM.,112İhtiyarA.,92İkizAÖ.,115İnanS.,67İşgörMM.,84,85,91İşgörenA.,37JJourneF.,40JungT.,42KKabadayıH.,49,108KahramanN.,56KahramanÖT.,51,121KalenderME.,112KanarE.,106KarademirB.,42,45,52KaraerT.,109KarakayaM.,113KaraözE.,117KarışM.,94KarlıtepeA.,56,110KaraoğluA.,57KaramanM.,57KaplanGS.,42KaplanPY.,104KaplancıklıZA.,131,133KasapB.,50KasapŞ.,50KayaS.,47KaynarAP.,111KeklikHM.,92KendirciR.,48,50,83,108

KılıçBA.,79,102KılıçE.,47KılınçK.,124,125KilitAC.,96KısakesenHİ.,121KıvrakG.,92KızılkayaP.,84KigC.,69KoçanM.,63KocatürkNM.,129,130KoçdorH.,57KoçdorMA.,57KoçBT.,78KoçtürkS.,45,52,54,82,89,126KoçyiğitS.,101,104KorkmazKS.,36KorkmazM.,66KoşarA.,130KurnazIA.,20KurtE.,60KurtFÖ.,48,87,98KurtN.,90KurtO.,87KurucaSE.,114KuşG.,65KutluA.,81KutluHM.,65,123,134KutluT.,93KüçükgülA.,84,85,91KüçükgüzelŞG.,101,104LLemkeG.,30LewED.,30LewalleP.,40MMaiorovEG.,69MansoubNH.,49MarwickJA.,30MenteşeA.,124MerimiM.,40MısırS.,61,124,125MiJ.,42MitouG.,69MocciaS.,28MusunuriS.,42


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NNagyA.,22NalbantA.,31,43,62,64NalbantsoyA.,75,94NurolNK.,119OOğuzE.,72,73OğuzoğluTÇ.,78OlgunA.,106OralÖ.,41,69,130OrunO.,101,104OğuzkanSB.,99ÖÖktemG.,120,135ÖnalT.,44,46,98ÖzaslanM.,99ÖzdemirA.,131,132ÖzdemirÖ.,76ÖzenB.,66ÖzenG.,54,126ÖzerNK.,45,60ÖzgerişFB.,88ÖzgülM.,67ÖzgüvenA.,83ÖzkanB.,92ÖzkutM.,87ÖzpolatB.,29,56,110ÖzsoyN.,114ÖztaşE.,81Öztopİ.,115ÖztuzcuS.,53,55,72,73,97,107ÖztürkO.,51,121ÖztürkZ.,80PPakM.,108PehlivanM.,55,99PekçetinÇ.,111PiacentiniM.,13PolatE.,88,90RRossiAG.,30

RouasR.,40RouhraziH.,120,135Russo,GL.,28RussoM.,28SSaatçioğluF.,19SagerÖ.,81SalkınH.,118SankoV.,77SaracaloğluA.,53,97SavaşHB.,70,71SaygılıN.,121SeçmeM.,59,122SerdarBS.,54,126SerinanE.,66SeverB.,131,133SeyhanMF.,51,121SeyrantepeV.,34SezerS.,77SezerÜA.,77SezermanOU.,69ShirwanH.,33SöğütMS.,20SökmenM.,55SönmezE.,74,76SözenE.,45SpagnuoloC.,28SüleymanoğluM.,114SütcüR.,92SüzgünPÇ.,101,104SweefO.,43ŞŞahinE.,20,127ŞahinYN.,88,90ŞanlıdağT.,63ŞekeroğluV.,79,102ŞekeroğluZA.,79,102ŞenerB.,53,97ŞentürkŞ.,18TTagaY.,52,82TaneliF.,83TanerF.,63TannenbaumSR.,38TanrıöverG.,67,100

TanrıverdiEÇ.,88,90TatarA.,74,76TedescoI.,28TekinC.,99TekirdağKA.,95TemizE.,53,55,97,107TiberPM.,101,104TimuçinED.,69TokgünO.,105TokgünPE.,105TopluN.,78TsengCY.,38TufanNLŞ.,59Tuğluİ.,87TuğluMİ.,44Turanİ.,124,125TurganN.,120,135TurhanNÖ.,50TürkçüÜÖ.,50TürkezH.,74,76TürkoğluC.,48,98UUğurMG.,72,73UğurlugüngörH.,106UlaşlıM.,53,97UluerET.,44,46,67,98UslusoyF.,71UysalA.,120,135UysalB.,81ÜÜnsalNP.,21VVandenabeeleP.,12VatanPÖ.,106VatanseverHS.,44,46,48,49,50,63,83,98,108VejselovaD.,65,123,134VuralSA.,37WWicherG.,42WoganGN.,38Y


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YalçınAS.,52,82YamanSÖ.,124,125YazıhanN.,47YerlikayaPO.,21YıldırımI.,93YıldızR.,83

YılmazAM.,45,52,82YılmazFM.,37YılmazO.,50YiğitA.,71YiğitB.,71YiğitZ.,91

YüceA.,41YumrutaşÖ.,55,107YumrutaşP.,55,107ZZhivotovskyB.,16

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