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)移演示文稿當(dāng)前1頁，總共49頁。（優(yōu)選）突變體能夠通過內(nèi)源性的促進(jìn)胰腺癌的轉(zhuǎn)移當(dāng)前2頁，總共49頁。BackgroundpreventsenescencePromotemetastasismutantp53drugresistanceInduceapoptosisp53antineoplasticpreventangiogenesis當(dāng)前3頁，總共49頁。Background75%p53mutanthighlymetastaticpoorprognosisdrugresistance當(dāng)前4頁，總共49頁。BackgroundKPCcell:stablelyexpresssmallhaipinRNA,lostremaningp53wide-typealle KPflCcell:ap53nullcelllineKPCmice:develophighlymetastaticpancreaticcancerthatfaithfullymimicsthehumandisease當(dāng)前5頁，總共49頁。IdeasMutantp53maintainmetastaticphenotypetherelationshipbetweenMutantp53andPDGFRbonmetastasisPDGFRbmadiatesmetastasiswhenMutantp53wasdepletedimatinibcaninhibitmetastasisbytargetingPDGFRbPDGFRbcorrelateswithdisease-freesurvival

PDGFRbasaDownstreamMediatorofMutantp53當(dāng)前6頁，總共49頁。Methods1.WoundHealingandInvasionAssays2.RNASequencingandDataAnalysis3.ImmunostainingandMicroscopy4.qRT-PCR5.PDGFRbLuciferaseReporterAssay6.CoimmunoprecipitationandChromatinImmunoprecipitation7.ImmunohistochemistryandImmunofluorescence8.MouseStudies當(dāng)前7頁，總共49頁。Questionwhether

mutantp53isneededtosustainthemetastaticphenotypeand

howitisregulated？當(dāng)前8頁，總共49頁。RESULTS當(dāng)前9頁，總共49頁。KPC+sh.Ctrlcellexpressmutantp53劃痕愈合率侵襲細(xì)胞數(shù)The

invasivenessdependon

mutantp53.Result1：SustainedExpressionofMutantp53IsRequiredforthe

InvasivePhenotypeofPancreaticCancerCells當(dāng)前10頁，總共49頁。轉(zhuǎn)移瘤數(shù)量whethermutantp53expressionwasrequiredto

sustainthemetastaticpotentialofKPCcells?轉(zhuǎn)移瘤熒光體視成像當(dāng)前11頁，總共49頁。Summary1theseresultsdemonstrate

that

mutantp53

cancontributeto

PDACinvasionandmetastasisandthatinhibitingitsactivitycanhaveanantimetastaticeffect當(dāng)前12頁，總共49頁。Questionhowmutantp53mediatestheinvasive

phenotypeofPDAC?當(dāng)前13頁，總共49頁。mutant

p53canaffecttheinvasivephenotypeofpancreaticcancer

cellsRNA測(cè)序，基因變化IPA分析Result2：TranscriptionalProfilingandFunctionalScreening

IdentifyPDGFRbasaDownstreamMediatorofMutant

p53inMurinePancreaticCancer當(dāng)前14頁，總共49頁。1：SLC40A12：SNED13：PDGFRbhypothesis：PDGFRbcouldaffectcellinvasionPDGFRb轉(zhuǎn)錄水平蛋白表達(dá)mutantp53敲除當(dāng)前15頁，總共49頁。細(xì)胞增殖不同亞型對(duì)侵襲的影響不同亞型蛋白表達(dá)情況當(dāng)前16頁，總共49頁。紅色熒光蛋白綠色熒光蛋白兩種細(xì)胞比值increasedPDGFRbdidnotconferaselectiveadvantagetotumorcellproliferation

當(dāng)前17頁，總共49頁。Summary2PDGFRbisnotrequired

fortheproliferationandtumorigenicpotentialofp53mutant

murinecancercellsbutspecificallyimpactstheirinvasive

potential.當(dāng)前18頁，總共49頁。QuestionIfthemutantp53-PDGFRbsignalingaxisacts

inhumancancercells？當(dāng)前19頁，總共49頁。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當(dāng)前20頁，總共49頁。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)胞當(dāng)前21頁，總共49頁。Summary3upregulationofthePDGFRb

isimportantfortheactionofmutantp53inPDACandpossibly

othertumortypes.當(dāng)前22頁，總共49頁。p73can

repressthetranscriptionof

PDGFRbourKPCcells

expressedp73,butnotp63whetherthephysicalinteractionofmutantp53withp73mightimpairtheabilityofp73to

negativelyregulatetheexpressionofPDGFRb?mutantp53caninhibitp73andp63Question當(dāng)前23頁，總共49頁。KPflCPDGFRb熒光素酶報(bào)告基因免疫共沉淀Result4：

Mutantp53Disruptsthep73/NF-YComplextoMediate

PDGFRbExpressionandTumorCellInvasion當(dāng)前24頁，總共49頁。howp73repressesPDGFRb

transcription?Question當(dāng)前25頁，總共49頁。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當(dāng)前26頁，總共49頁。whethertherepressiveactionofp73on

PDGFRbtranscriptionwasmediatedbyNF-Yandmodulated

bymutantp53?Question當(dāng)前27頁，總共49頁。KPflC細(xì)胞侵襲能力PDGFRb熒光素酶報(bào)告基因當(dāng)前28頁，總共49頁。KPflC細(xì)胞KPC侵襲能力修復(fù)當(dāng)前29頁，總共49頁。Summary4mutantp53promotesinvasion,inpart,dependsonitsabilityto

enhancePDGFRbexpressionthroughthedisruptionoftheinhibitoryp73/NF-Ycomplex當(dāng)前30頁，總共49頁。whetherPDGFRblevels

regulatemetastaticbehaviorofPDAC

cellsinmice?Question當(dāng)前31頁，總共49頁。肺轉(zhuǎn)移肺組織病理切片Result5：

ModulationofPDGFRbExpressionLevelsMediatesthe

PhenotypicEffectsofMutantp53DepletionInVivo當(dāng)前32頁，總共49頁。轉(zhuǎn)移灶大小免疫組化whetherpharmacologicinhibition

ofthePDGFRbpathwayrecapitulatestheeffectsofPDGFRbor

mutantp53depletion?當(dāng)前33頁，總共49頁。時(shí)效關(guān)系實(shí)驗(yàn)量效關(guān)系實(shí)驗(yàn)Crenolanib兩種細(xì)胞IC50當(dāng)前34頁，總共49頁。whethercrenolanibcansuppressmetastasis?Crenolanib處理與DMSO處理熒光和免疫組化細(xì)胞凋亡當(dāng)前35頁，總共49頁。KPflC細(xì)胞pancreaticcancercellscanprovideasourceofPDGFligandthatcould

triggerautocrineactivationofPDGFRb條件培養(yǎng)當(dāng)前36頁，總共49頁。Summary5abrogationof

autocrineactivationsignalingofPDGFRb

leadstoasignificantreductionofinvasionandmetastasisdrivenbymutantp53.當(dāng)前37頁，總共49頁。whetherPDGFRbinhibitionpreventsthedevelopmentofmetastasisinKPCmice?

Question當(dāng)前38頁，總共49頁。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(yàn)Imatinib處理KPC的肺轉(zhuǎn)移Result5：

ImatinibInhibitstheDevelopmentofMetastasesina

PDACMouseModelbyTargetingPDGFRb當(dāng)前39頁，總共49頁。thehighdiseaseburdeninthepancreaswastheprimarycauseofdeath腫瘤體積總體存活率當(dāng)前40頁，總共49頁。92%15%其他器官轉(zhuǎn)移情況不同器官HE染色當(dāng)前41頁，總共49頁。DAPI,blue;CK8,red;pPDGFRb,green

imatinibwasabletoeffectively

inhibitPDGFRbactivityinprimarytumorsbasedonreduced

levelsofphospho-PDGFRb

inthetumorcellspPDGFRb強(qiáng)度imatinib對(duì)體內(nèi)PDGFRb的影響當(dāng)前42頁，總共49頁。Summary6byinhibitingthekinase

activityofPDGFRb,imatinibsignificantlydiminishesthemetastaticpotentialofpancreaticcancercells.當(dāng)前43頁，總共49頁。whetherupregulationofPDGFRbiscorrelatedwithprognosisorwiththeclinicopathologicalcharacteristicsofPDACsinpatients.Ques