突變體能夠通過內(nèi)源性的促進胰腺癌的轉移演示文稿

突變體能夠通過內(nèi)源性的促進胰腺癌的轉移演示文稿現(xiàn)在是1頁\一共有49頁\編輯于星期日（優(yōu)選）突變體能夠通過內(nèi)源性的促進胰腺癌的轉移現(xiàn)在是2頁\一共有49頁\編輯于星期日BackgroundpreventsenescencePromotemetastasismutantp53drugresistanceInduceapoptosisp53antineoplasticpreventangiogenesis現(xiàn)在是3頁\一共有49頁\編輯于星期日Background75%p53mutanthighlymetastaticpoorprognosisdrugresistance現(xiàn)在是4頁\一共有49頁\編輯于星期日BackgroundKPCcell:stablelyexpresssmallhaipinRNA,lostremaningp53wide-typealle KPflCcell:ap53nullcelllineKPCmice:develophighlymetastaticpancreaticcancerthatfaithfullymimicsthehumandisease現(xiàn)在是5頁\一共有49頁\編輯于星期日IdeasMutantp53maintainmetastaticphenotypetherelationshipbetweenMutantp53andPDGFRbonmetastasisPDGFRbmadiatesmetastasiswhenMutantp53wasdepletedimatinibcaninhibitmetastasisbytargetingPDGFRbPDGFRbcorrelateswithdisease-freesurvival

PDGFRbasaDownstreamMediatorofMutantp53現(xiàn)在是6頁\一共有49頁\編輯于星期日Methods1.WoundHealingandInvasionAssays2.RNASequencingandDataAnalysis3.ImmunostainingandMicroscopy4.qRT-PCR5.PDGFRbLuciferaseReporterAssay6.CoimmunoprecipitationandChromatinImmunoprecipitation7.ImmunohistochemistryandImmunofluorescence8.MouseStudies現(xiàn)在是7頁\一共有49頁\編輯于星期日Questionwhether

mutantp53isneededtosustainthemetastaticphenotypeand

howitisregulated？現(xiàn)在是8頁\一共有49頁\編輯于星期日RESULTS現(xiàn)在是9頁\一共有49頁\編輯于星期日KPC+sh.Ctrlcellexpressmutantp53劃痕愈合率侵襲細胞數(shù)The

invasivenessdependon

mutantp53.Result1：SustainedExpressionofMutantp53IsRequiredforthe

InvasivePhenotypeofPancreaticCancerCells現(xiàn)在是10頁\一共有49頁\編輯于星期日轉移瘤數(shù)量whethermutantp53expressionwasrequiredto

sustainthemetastaticpotentialofKPCcells?轉移瘤熒光體視成像現(xiàn)在是11頁\一共有49頁\編輯于星期日Summary1theseresultsdemonstrate

that

mutantp53

cancontributeto

PDACinvasionandmetastasisandthatinhibitingitsactivitycanhaveanantimetastaticeffect現(xiàn)在是12頁\一共有49頁\編輯于星期日Questionhowmutantp53mediatestheinvasive

phenotypeofPDAC?現(xiàn)在是13頁\一共有49頁\編輯于星期日mutant

p53canaffecttheinvasivephenotypeofpancreaticcancer

cellsRNA測序，基因變化IPA分析Result2：TranscriptionalProfilingandFunctionalScreening

IdentifyPDGFRbasaDownstreamMediatorofMutant

p53inMurinePancreaticCancer現(xiàn)在是14頁\一共有49頁\編輯于星期日1：SLC40A12：SNED13：PDGFRbhypothesis：PDGFRbcouldaffectcellinvasionPDGFRb轉錄水平蛋白表達mutantp53敲除現(xiàn)在是15頁\一共有49頁\編輯于星期日細胞增殖不同亞型對侵襲的影響不同亞型蛋白表達情況現(xiàn)在是16頁\一共有49頁\編輯于星期日紅色熒光蛋白綠色熒光蛋白兩種細胞比值increasedPDGFRbdidnotconferaselectiveadvantagetotumorcellproliferation

現(xiàn)在是17頁\一共有49頁\編輯于星期日Summary2PDGFRbisnotrequired

fortheproliferationandtumorigenicpotentialofp53mutant

murinecancercellsbutspecificallyimpactstheirinvasive

potential.現(xiàn)在是18頁\一共有49頁\編輯于星期日QuestionIfthemutantp53-PDGFRbsignalingaxisacts

inhumancancercells？現(xiàn)在是19頁\一共有49頁\編輯于星期日PDGFRbmRNA水平四種人胰腺癌細胞（Mutp53敲除）不同種類腫瘤中Mutp53敲除Result3：PDGFRbMediates

Mutantp53ActioninHumanCancer

Cells現(xiàn)在是20頁\一共有49頁\編輯于星期日mutp53/PDGFRb分別敲除，A2.1細胞侵襲情況p53/PDGFRb敲除p53-/-細胞過表達PDGFRbinvasionp53-/-人胰腺癌細胞現(xiàn)在是21頁\一共有49頁\編輯于星期日Summary3upregulationofthePDGFRb

isimportantfortheactionofmutantp53inPDACandpossibly

othertumortypes.現(xiàn)在是22頁\一共有49頁\編輯于星期日p73can

repressthetranscriptionof

PDGFRbourKPCcells

expressedp73,butnotp63whetherthephysicalinteractionofmutantp53withp73mightimpairtheabilityofp73to

negativelyregulatetheexpressionofPDGFRb?mutantp53caninhibitp73andp63Question現(xiàn)在是23頁\一共有49頁\編輯于星期日KPflCPDGFRb熒光素酶報告基因免疫共沉淀Result4：

Mutantp53Disruptsthep73/NF-YComplextoMediate

PDGFRbExpressionandTumorCellInvasion現(xiàn)在是24頁\一共有49頁\編輯于星期日howp73repressesPDGFRb

transcription?Question現(xiàn)在是25頁\一共有49頁\編輯于星期日p73NF-YBPDGFRb序列×√NF-Y

bindingto

thePDGFRBpromoterp73/NF-Y

interactionhampersNF-YtobindandactivatethePDGFRB

promoterp73bindingwithNF-YBmutantp53canaffectp73/NF-YBPDGFRb結合定量×NF-YBNF-YANF-YC現(xiàn)在是26頁\一共有49頁\編輯于星期日whethertherepressiveactionofp73on

PDGFRbtranscriptionwasmediatedbyNF-Yandmodulated

bymutantp53?Question現(xiàn)在是27頁\一共有49頁\編輯于星期日KPflC細胞侵襲能力PDGFRb熒光素酶報告基因現(xiàn)在是28頁\一共有49頁\編輯于星期日KPflC細胞KPC侵襲能力修復現(xiàn)在是29頁\一共有49頁\編輯于星期日Summary4mutantp53promotesinvasion,inpart,dependsonitsabilityto

enhancePDGFRbexpressionthroughthedisruptionoftheinhibitoryp73/NF-Ycomplex現(xiàn)在是30頁\一共有49頁\編輯于星期日whetherPDGFRblevels

regulatemetastaticbehaviorofPDAC

cellsinmice?Question現(xiàn)在是31頁\一共有49頁\編輯于星期日肺轉移肺組織病理切片Result5：

ModulationofPDGFRbExpressionLevelsMediatesthe

PhenotypicEffectsofMutantp53DepletionInVivo現(xiàn)在是32頁\一共有49頁\編輯于星期日轉移灶大小免疫組化whetherpharmacologicinhibition

ofthePDGFRbpathwayrecapitulatestheeffectsofPDGFRbor

mutantp53depletion?現(xiàn)在是33頁\一共有49頁\編輯于星期日時效關系實驗量效關系實驗Crenolanib兩種細胞IC50現(xiàn)在是34頁\一共有49頁\編輯于星期日whethercrenolanibcansuppressmetastasis?Crenolanib處理與DMSO處理熒光和免疫組化細胞凋亡現(xiàn)在是35頁\一共有49頁\編輯于星期日KPflC細胞pancreaticcancercellscanprovideasourceofPDGFligandthatcould

triggerautocrineactivationofPDGFRb條件培養(yǎng)現(xiàn)在是36頁\一共有49頁\編輯于星期日Summary5abrogationof

autocrineactivationsignalingofPDGFRb

leadstoasignificantreductionofinvasionandmetastasisdrivenbymutantp53.現(xiàn)在是37頁\一共有49頁\編輯于星期日whetherPDGFRbinhibitionpreventsthedevelopmentofmetastasisinKPCmice?

Question現(xiàn)在是38頁\一共有49頁\編輯于星期日量效關系抑制細胞增殖所需IC50體外KPC細胞侵襲實驗Imatinib處理KPC的肺轉移Result5：

ImatinibInhibitstheDevelopmentofMetastasesina

PDACMouseModelbyTargetingPDGFRb現(xiàn)在是39頁\一共有49頁\編輯于星期日thehighdiseaseburdeninthepancreaswastheprimarycauseofdeath腫瘤體積總體存活率現(xiàn)在是40頁\一共有49頁\編輯于星期日92%15%其他器官轉移情況不同器官HE染色現(xiàn)在是41頁\一共有49頁\編輯于星期日DAPI,blue;CK8,red;pPDGFRb,green

imatinibwasabletoeffectively

inhibitPDGFRbactivityinprimarytumorsbasedonreduced

levelsofphospho-PDGFRb

inthetumorcellspPDGFRb強度imatinib對體內(nèi)PDGFRb的影響現(xiàn)在是42頁\一共有49頁\編輯于星期日Summary6byinhibitingthekinase

activityofPDGFRb,imatinibsignificantlydiminishesthemetastaticpotentialofpancreaticcancercells.現(xiàn)在是43頁\一共有49頁\編輯于星期日whetherupregulationofPDGFRbiscorrelatedwithprognosisorwiththeclinicopathologicalcharacteristicsofPDACsinpatients.Question現(xiàn)在是4