1EpigeneticsilencingofmiRNA-338-5pandmiRNA-421drivesSPINK1-positiveprostatecancerVipulBhatia1,5,AnjaliYadav1,5,RitikaTiwari1,ShivanshNigam1,SakshiGoel1,ShannonCarskadon2,NileshGupta3,ApulGoel4,NallasivamPalanisamy2,BushraAteeq1*1MolecularOncologyLab,DepartmentofBiologicalSciencesandBioengineering,IndianInstituteofTechnologyKanpur,Kanpur,208016,U.P.,INDIA2VattikutiUrologyInstitute,DepartmentofUrology,HenryFordHealthSystem,Detroit,MI48202,USA3DepartmentofPathology,HenryFordHealthSystem,Detroit,MI48202,USA4DepartmentofUrology,KingGeorge'sMedicalUniversity,Lucknow,226003,U.P.,INDIA5Theseauthorscontributedequallytothiswork.RunningTitle:SilencingofmiR-338-5p/-421drivesSPINK1-positivecancer.Financialsupport:ThisworkissupportedbytheWellcomeTrust/DBTIndiaAlliancegrant(IA/I(S)/12/2/500635toBA).Conflictofinterest:Theauthorsdeclarenoconflictsofinterestordisclosures.*CorrespondingAuthorandLeadContact:BushraAteeq,MolecularOncologyLab,DepartmentofBiologicalSciencesandBioengineering,IndianInstituteofTechnologyKanpur,Kanpur,208016,U.P.,INDIAPhone:+915122594083,Fax:+915122594010Email:bushra@iitk.ac.in(B.Ateeq)CancerResearch.onDecember26,2018.©2018AmericanAssociationforclincancerres.aacrjournals.orgDownloadedfromAuthormanuscriptshavebeenpeerreviewedandacceptedforpublicationbuthavenotyetbeenedited.AuthorManuscriptPublishedOnlineFirstonDecember26,2018;DOI:10.1158/1078-0432.CCR-18-32302AbstractPurpose:SerinePeptidaseInhibitor,Kazaltype-1(SPINK1)overexpressiondefinesthesecondmostrecurrentandaggressiveprostatecancer(PCa)subtype.However,theunderlyingmolecularmechanismandpathobiologyofSPINK1inPCaremainslargelyunknown.ExperimentalDesign:MicroRNA-predictiontoolswereemployedtoexaminetheSPINK1-3’UTRformiRNAsbinding.LuciferasereporterassayswereperformedtoconfirmtheSPINK1-3’UTRbindingofshortlistedmiR-338-5p/miR-421.Further,miR-338-5p/-421overexpressingcancercells(SPINK1-positive)wereevaluatedforoncogenicpropertiesusingcell-basedfunctionalassaysandmicexenograftmodel.Globalgeneexpressionprofil...
