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DoctoralCandidatesEntrance

Examinationof

*****UniversityCandidate:******ApplyingMajor:MicrobialandbiochemicalpharmacyOutlineEducationExperience

ResearchExperiencePublicationsMasterResearchDoctoralResearchPlanEducationExperiencePeriodMajordegreeSchool2011.09-2015.07BiologicalEngineeringbachelordegree***2015.09-MicrobialandbiochemicalpharmacyMasterdegree****EducationExperienceTraditionalChineseMedicineBureauofGuangdongProvince(NO.20141065),NaturalScienceFoundationofGuangdongProvince(NO.2017A32217007)NationalUniversityStudentInnovationProgram.ProfessionalThird-classScholarshipfrom2015-2018academicyearsAwardsPublications******,

EnzeYang,LianghuaShen,DeweiNiu,Mason

Breitzig,LeeCharlesTan,FengWang*.Thenoveltruncatedisoformofhumanmanganesesuperoxidedismutasehasadifferentialroleinpromotingmetastasisoflungcancercells.CellBiologyInternational.

Accepted.doi:10.1002/cbin.10972.******,

LianghuaShen,SijiaLei,Mason

Breitzig,LeeCharlesTan,QingZheng,HanxiaoSun,FengWang*.PTBP1enhancesexon11askippinginMenapre-mRNAtopromotemigrationandinvasioninlungcarcinomacells.TheJournalofBiologicalChemistry.Majorrevision.JBC/2018/002627R1.DeweiNiu,******,ShanzeYi,FengWang*.Genecloning,proteinexpressionandfunctionalanalysisofatype3metallothioneingenefromSonneratiaalbawithbiosorptionpotential.PolishJournalofEnvironmentalStudies,Accepted.PJOES-00647-2017-02.MasterResearchPTBP1enhancesexon11askippinginMenapre-mRNAtopromotemigrationandinvasioninlungcarcinomacellsBackgroundObjectivesTechnologyMethodsResultsConclusionsBackgroundSevenalternativesplicing(AS)subgroups:Exonskippingaccountsfornearly40%ofASevents;alternative3′splicesite(3′SS)selection(18.4%)and5′SSselection(7.9%);intronretentionaccountsforlessthan5%;mutuallyexclusiveexons,alternativepromoterusage(3′UTR)andalternativepolyadenylation(5′UTR)arenotfrequent.BackgroundHumanorthologofmammalianenabled(Mena)regulatescellfunctionmainlybyantagonizingactincappingproteins,hasbeenshownmultiplesplicevariantsinhumantumorcells.MenaINVforinvasivefeatureMena11aforepithelial-specificfeatureBackgroundTwomainASregulatorsheterogeneousnuclearribonucleoprotein(hnRNPs)andserine/arginine-rich(SR)proteins.PolypyrimidineTract-BindingProtein1(PTBP1,alsoknownashnRNPI)bindspreferentiallytopyrimidine-richsequences.

ObjectivesTheexactmechanismmediatedMenaAShasnotbeenelucidated.ThepurposeofthisstudyistoexplorethemolecularmechanismofPTBP1inregulatingMenaalternativesplicingandroleofPTBP1inlungcarcinomacellsmetastasis.RegulatedbyPTBP1?TechnologyMethods

ResultsPTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesand95-DcellsandupregulationofPTBP1isassociatedwithEMTprogressPTBP1promotesmigrationandinvasionoflungcancercellsPTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercellsPTBP1enhancesalternativesplicingofexon11athroughitsinteractionwithbothupstreamanddownstreamintronsequencesPTBP1-mediatedmigrationandinvasionof95-DcellsarepartiallydependentonMenaINV

Results1.1.PTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesA.PTBP1expressionwaselevatedinLUADtissues(N=515)comparedwithnormallungtissues(N=59)accordingtoTCGAdatabase(p<0.01);B.Kaplan-MeierplotsofpatientswithLUADaccordingtohigh(N=127)andlow(N=375)PTBP1expressionfromtheTCGAdatabaseandcomparedbypairedt-test,p<0.01.AB

Results1.2.PTBP1ishighlyexpressedinmetastaticlungcancercells(95-D)

Results1.3.UpregulationofPTBP1isassociatedwithTGFβinducedEMTprogress

ResultsPTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesand95-DcellsandupregulationofPTBP1isassociatedwithEMTprogressPTBP1promotesmigrationandinvasionoflungcancercellsPTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercellsPTBP1enhancesalternativesplicingofexon11athroughitsinteractionwithbothupstreamanddownstreamintronsequencesPTBP1-mediatedmigrationandinvasionof95-DcellsarepartiallydependentonMenaINV

Results2.1.OverexpressedPTBP1promoteslevelsofEMT-relatedproteinsinlungcancercells

Results2.2.OverexpressedofPTBP1promotesmigrationoflungcancercells

Results2.3.KnockdownofPTBP1inhibitslevelsofEMT-relatedproteinsinlungcancercells

Results2.4.KnockdownofPTBP1inhibitsmigrationoflungcancercells

Results2.5.PTBP1promotesinvasionoflungcancercells

ResultsPTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesand95-DcellsandupregulationofPTBP1isassociatedwithEMTprogressPTBP1promotesmigrationandinvasionoflungcancercellsPTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercellsPTBP1enhancesalternativesplicingofexon11athroughitsinteractionwithbothupstreamanddownstreamintronsequencesPTBP1-mediatedmigrationandinvasionof95-DcellsarepartiallydependentonMenaINV

Results3.1.MenaINV

iselevatedinthehighlymetastaticlungcancercellsAB

Results3.2.PTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercells.

ResultsPTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesand95-DcellsandupregulationofPTBP1isassociatedwithEMTprogressPTBP1promotesmigrationandinvasionoflungcancercellsPTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercellsPTBP1enhancesalternativesplicingofexon11athroughitsinteractionwithbothupstreamanddownstreamintronsequencesPTBP1-mediatedmigrationandinvasionof95-DcellsarepartiallydependentonMenaINV

Results4.1.DiagramoftheMenapre-mRNAminigenespanningfromexon10toexon12B

Results4.2.PTBP1enhancesexon11askippinginMenaminigene

Results4.3.PTBP1bindstobothupstreamanddownstreamintronsequencesinMenapre-mRNABCA.SchemeofwildMenaminigeneandmutantMenaminigene.TheunderlinedsequencesareconsensusPTBP1bindingsites.B.RT-PCRassayswereperformedtodetectthechangesofexon11askippingofwildandmutantMenaminigenesinPTBP1overexpressedA549cells.

C.WesternblotanalysisofRNA-pulldownassaysshowedthatthechangesofinteractionsbetweenPTBP1andbiotin-RNAoligomerscontainingpotentialPTBP1bindingsites.

ResultsPTBP1ishighlyexpressedinlungadenocarcinoma(LUAD)tissuesand95-DcellsandupregulationofPTBP1isassociatedwithEMTprogressPTBP1promotesmigrationandinvasionoflungcancercellsPTBP1enhancesexon11askippingofMenapre-mRNAinlungcancercellsPTBP1enhancesalternativesplicingofexon11athroughitsinteractionwithbothupstreamanddownstreamintronsequencesPTBP1-mediatedmigrationandinvasionof95-DcellsarepartiallydependentonMenaINV

Results5.1.PTBP1-mediatedmigrationof95-DcellsispartiallydependentonMenaINV

Results5.3.ThefilopodiaformationwasregulatedbyPTBP1anddependedonchangeofMenaINV.Thefieldsofviewswerecapturedat10×40magnification(Scalebars:10μm)

ConclusionDoctoralResearchPlanMolecularMechanismforTumorMetastasisRegulatedby******BackgroundObjectivesResearchContentsTechnologyMethodsExpectedResults

Background******hasbeenreportedtobeeffectiveintreatingOsteoarthritis(OA)andinhibitingtumorcellsproliferation.Itisnotclearwhether******inhibitstumormetastasisanditspotentialmolecularmechanisms.ObjectivesTostudytheeffect******ontheregulating

tumormetastasisinseveralcancercells.Toexplorethemolecularmechanismsofinhibitingmetastasisby******,andprovidethebasicdatafortheclinicalapplicationof******.ResearchContentsThemetastaticmodeinvitro

isinducedbytransforminggrowthfactor-β(TGF-β).Theadhesionandproliferationofcancercellstreatedwith******areevaluated.Wound-healingandtranswellassaysareperformedtostudytheeffect******

ontheregulating

tumormigrationandinvasion.Thetumormetastasismodeinvivoisestablishedinnudemouse.TranscriptomeandProteomicsareusedtodetecttheexpressionsoftranscriptionfactorsorpathway-relatedproteins.Inhibitorsareusedtodemonstratethemolecularmechanismsofmetastasisinhibitedby******.TechnologyMethodsMolecularMechanismforTumorMetastasisRegulatedbyXanthanGumRegulationofTumorMetastasisStudyofthemolecularmechanismInvitroInvivoCellviabilityCellcycleCelladhesionMigrationInvasionTumorgrowthTumormetastasisImmunohistochemistryVivoimagingTranscriptomeProteomicsWesternblotqPCRInhibitorImmunofluorescence******ExpectedResults

******caneffectivelyinhibitthe