突變體能夠通過內(nèi)源性的促進(jìn)胰腺癌的轉(zhuǎn)移演示文稿

突變體能夠通過內(nèi)源性的促進(jìn)胰腺癌的轉(zhuǎn)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(jìn)胰腺癌的轉(zhuǎn)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(xì)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(zhuǎn)移瘤數(shù)量whethermutantp53expressionwasrequiredto

sustainthemetastaticpotentialofKPCcells?轉(zhuǎn)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(zhuǎn)錄水平蛋白表達(dá)mutantp53敲除現(xiàn)在是15頁\一共有49頁\編輯于星期日細(xì)胞增殖不同亞型對侵襲的影響不同亞型蛋白表達(dá)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(xì)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(xì)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(xì)胞侵襲情況p53/PDGFRb敲除p53-/-細(xì)胞過表達(dá)PDGFRbinvasionp53-/-人胰腺癌細(xì)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×√N(yùn)F-Y

bindingto

thePDGFRBpromoterp73/NF-Y

interactionhampersNF-YtobindandactivatethePDGFRB

promoterp73bindingwithNF-YBmutantp53canaffectp73/NF-YBPDGFRb結(jié)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(xì)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(xì)胞KPC侵襲能力修復(fù)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(zhuǎn)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(zhuǎn)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(guān)系實(shí)驗量效關(guān)系實(shí)驗Crenolanib兩種細(xì)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(xì)胞凋亡現(xiàn)在是35頁\一共有49頁\編輯于星期日KPflC細(xì)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(guān)系抑制細(xì)胞增殖所需IC50體外KPC細(xì)胞侵襲實(shí)驗Imatinib處理KPC的肺轉(zhuǎn)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(zhuǎn)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(qiáng)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