cisplatinchemotherapyonhumanovarycancercel

EFFECTOFMIFEPRISTONEUSEDALONEORWITHCISPLATINCHEMOTHERAPYONHUMANOVARIANCANCERCELLGROWTHByELIZABETHMCKAYFREEBURGB.S.,UniversityofNotreDame,NotreDame,Indiana,2002M.A.,UniversityofSouthDakota,Vermillion,SouthDakota,2003ADissertationSubmittedinPartialFulfillmentofTheRequirementsfortheDegreeofDoctorofPhilosophyDivisionofBasicBiomedicalSciencesIntheGraduateSchoolSanfordSchoolofMedicineofTheUniversityofSouthDakotaDecember,2008UMINumber:3351178INFORMATIONTOUSERSThequalityofthisreproductionisdependentuponthequalityofthecopysubmitted.Brokenorindistinctprint,coloredorpoorqualityillustrationsandphotographs,printbleed-through,substandardmargins,andimproperalignmentcanadverselyaffectreproduction.Intheunlikelyeventthattheauthordidnotsendacompletemanuscriptandtherearemissingpages,thesewillbenoted.Also,ifunauthorizedcopyrightmaterialhadtoberemoved,anotewillindicatethedeletion.®UMIUMIMicroform3351178Copyright2009byProQuestLLC.Allrightsreserved.ThismicroformeditionisprotectedagainstunauthorizedcopyingunderTitle17,UnitedStatesCode.ProQuestLLC789E.EisenhowerParkwayPOBox1346AnnArbor,Ml48106-1346ThemembersofthecommitteeappointedtoexaminethedissertationofElizabethMcKayFreeburgfinditsatisfactoryandrecommendthatitbeaccepted.Chairperson,CarlosM.Teller/a,Ph.D&MU,«W\ZTJSL^LKathleenM.Ey&ter,Ph.DKristiA.Egland^h.DDo&glasS.Martin,Ph.D.WKeithMfekimins,Ph.D.%MJM^_^^A//EvelynH^Schlenker,Ph.D.11ABSTRACTOvariancancer(OvCa)causesmoredeathsthananyothergynecologicmalignancy.Itisusuallydiagnosedatlatestagesandpatientsrequirecytoreductivesurgeryfollowedbyplatinum-basedchemotherapy.However,theefficacyofplatinumderivativesishinderedbytheirelevatedtoxicity,thedevelopmentoftumortolerancetothedrugs,andthere-growthorrepopulationoftumorcellsbetweentreatmentintervals.Hence,searchfornewmeasurestoenhancetheefficacyofplatinumderivativesforOvCaisimperative.Preclinicalstudiesinourlaboratoryhavedemonstratedthattheantiprogestinsteroid,mifepristone(MF),iseffectiveasasingleagentinvitroandinvivoinhibitinggrowthofhumanepithelialOvCacells.MFinducesareversibleblockageofthecellcycleattheGl-to-Stransitionandastronginhibitionofcyclindependentkinase2(Cdk2)activity.BasedonthesedataandthefactthatCdk2isfrequentlyup-regulatedinovariantumors,thisprojectwasdevelopedtotestthehypothesisthatMFenhancestheefficacyofplatinum-basedtherapy-utilizingcisplatin(CDDP)asaprototype.First,wesoughttodetermineifthecytostaticconcentrationsofMFpreventsrepopulationofOvCacellsoccurringbetweenCDDPtreatmentintervals.EvidenceispresentedonthegenerationofaninvitromodelofOvCarepopulationbetweenCDDProunds,itsefficientabrogationbyMF,andtheapparentenhancementofCDDP-inducedtoxicitybyMF.Secondly,weaskedwhetherthetoxicityofMFoccurredregardlessofthedegreeofsensitivitydevelopedbytheOvCacellstoCDDP,andofthestatusofthep53tumorsuppressorgene.EvidenceispresentedsuggestingthatMF-inducedcytotoxicityinvolvingcytostasisatlowconcentrations,andcaspase-associatedapoptoticlethalityathighconcentrations,doesnotcorrelatewiththesensitivityofthecellstoCDDP,andappearstobeindependentofthep53geneticstatus.Overall,ourstudiesprovideevidencetosuggestthatMFusedeitheraloneortogetherwithCDDPhasthepotentialtoimproveOvCatreatmentsuccess.UsingMFtherapyinassociationwith,andafterplatinumchemotherapy,maysignificantlyimprovethe5yearsurvivalrateforOvCa,andconceivablyconvertthislethalcancerintoatreatablechronicdisease.CartosM.Telleria,Ph.D.ResearchAdvisorinACKNOWLEGEMENTSFirstandforemost,Ithankmyresearchmentor,Dr.CarlosTelleria,forhisguidance,support,andencouragementasIconductedmystudiesinhislaboratory.Carloshasspentcountlesshoursworkingtoshapemeintoacriticalthinker,aconfidentandconsistentpresenter,andabetterwriter.IthankCarlosforthetimeandenergyhedevotedtomesothatIcouldsucceed.IappreciatethemanycoffeebreaksandlivelyphilosophicalandpoliticaltalksIhadwithhiminthelabovertheyears.Heisagreatfriendandmentor.Iadmirehishard-working,persistentpersonality.ThankstoCarlos.Withouthishelpandguidanceoverthelastfourandahalfyears,thisprojectwouldnothavebeenpossible.Secondly,Ithankmyresearchcolleagueandfriend,Dr.AliciaGoyeneche,forherconstantsupportandhelp.Shetaughtmemanyofthetechnicalaspectsofthelaboratoryandtaughtmetopayattentiontodetails.Aliciawasthereformeconstantly.Ithankherfortheassistanceoncompletingmyrepopulationexperiments.Aliciahasacontagiouspassionforresearchwhichhasmademeenthusiasticaboutresearch.Aliciaisawonderfulmomtoherchildrenandhasbeenagreatexampletomeofbalancingworklifewithfamilylife.Iwillalwaysrememberthestorieswehavesharedaboutlifeandourfamiliesandlookforwardtosharingmorestoriesinthefuture.Together,CarlosandAliciahavecreatedaPh.D.experienceformethatcouldnothavebeenanybetter!Ithankthemforbeingsuchgreatrolemodelstomeandfortakingmeundertheirwingstomakemeasolidresearcher.TheTellerialaboratoryisaspecialplacetobepartofbecauseitfunctionslikeabigfamily.Eachpersonworkstogetherforacommongoalandeveryonebenefitsbecauseofit.Thisterrificenvironmentwouldnothavebeencreatedwithoutthehelpofmycurrentlabmates,StacyRowenhorstandivChelseaTieszen,andmypastlabmates,Eri