《肿瘤细胞信号转导》PPT课件

如何阅读文献,在肿瘤学方面有哪些好的杂志如何快速阅读文献以获取知识论文的主要结构,肿瘤的分子信号转导,+genomicinstability,fromHanahanandWeinberg2000,SignalTransductionandCancer,LectureI:GrowthFactorsandReceptorsOutline:WhatisSignalTransduction?WhatareGrowthFactors?HowdotheycontributetonormalST?HowisthisSTderegulatedinCancer?,LectureI:GrowthFactorsandReceptorsWhatisSignalTransduction?SignalTransductionistheprocessbywhichacellconvertsanextracellularsignalintoaresponse.Involvedin:Cell-cellcommunicationCellsresponsetoenvironmentIntracellularhomeostatsis-internalcommunication,GenericSignallingPathway,SignalReceptor(sensor)TransductionCascadeTargetsResponse,AlteredMetabolism,MetabolicEnzyme,GeneRegulator,CytoskeletalProtein,AlteredGeneExpression,AlteredCellShapeorMotility,AdaptedfromMolecularBiologyoftheCell,(2002),4thedition,Albertsetal.,ComponentsofSignalling,WhatcanbetheSignal?Externalmessagetothecell,Peptides/Proteins-GrowthFactorsAminoacidderivatives-epinephrine,histamineOthersmallbiomolecules-ATPSteroids,prostaglandinsGases-NitricOxide(NO)PhotonsDamagedDNAOdorants,tastants,Signal=LIGANDLigand-Amoleculethatbindstoaspecificsiteonanothermolecule,usuallyaprotein,iereceptor,ComponentsofSignalling,WhatareReceptors?Sensors,whatthesignal/ligandbindstoinitiateST,CellsurfaceIntracellular,HydrophillicLigand,Cell-SurfaceReceptor,Plasmamembrane,HydrophobicLigand,CarrierProtein,IntracellularReceptor,Nucleus,AdaptedfromMolecularBiologyoftheCell,(2002),4thedition,Albertsetal.,CellSurfaceReceptorTypes:Ligand-gatedionchannel,CellSurfaceReceptorTypes:2)G-ProteinCoupledReceptor,CellSurfaceReceptorTypes:3)Enzyme-linkedReceptoregGrowthFactorReceptors,GrowthFactors,LigandswhichbindenzymelinkedreceptorsSignaldiversecellularresponsesincluding:ProliferationDifferentiationGrowthSurvivalAngiogenesisCansignaltomultiplecelltypesorbespecific,GrowthFactorReceptors,MostgrowthfactorsbindReceptorTyrosineKinases,GrowthFactorReceptorActivationI,RTK,RS/TK,GrowthFactorReceptorActivationII,Growthsignalautonomy,Insensitivitytoanti-growthsignals,Resistancetoapoptosis:Uncouplecellsgrowthprogramfromsignalsintheenvironment.Growthfactorsinnormalcellsserveasenvironmentalsignals.,GrowthFactorSTandCancer,Growthfactorsregulategrowth,proliferation,andsurvival.Theseareallderegulatedincancer.,HanahanandWeinberg,(2000)HallmarksofCancer,Cell(100)57,GrowthfactorswithOncogenicPotential,PDGF,originallyshowntoregulateproliferation,wasalsoshowntohavehomologytov-sis,thesimiansarcomavirus.OtherviraloncogenesencodedproteinproductsthatweregrowthfactorsthatoftenoverexpressedincancersuchasTGF-a.Autocrinesignallingleadstoderegulatedgrowth.,PDGFfamilyNeurotrophinsAchainNGFBchain(c-sis)BDNFFGFFamilyNT3acidicFGFCytokines(Hematopoietic)basicFGFIL-2EGFFamilyIL-3EGFM-CSFTGF-aGM-CSF,GFReceptorswithOncogenicPotential,EGFR,kinaseactivitystimulatedbyEGF-1andTGF-ainvolvedincellgrowthanddifferentiation,waslinkedviasequencehomologytoaknownavianerythroblastosisvirusonocgene,v-erbB.Sincethen,manyoncogeneshavebeenshowntoencodeforGFRs.,EGFRfamilyInsulinReceptorfamilyerbB1(c-erbB)IGF-1(c-ros)erbB2(neu)NeurotrophinsFGFFamilyNGFR(trk)FGFR-1(fig)BDNFR(trk-B)FGFR-2(K-sam)NT3R(trk-C)PDGFRFamilyCSF-1R(c-fms)SLFR(c-kit),Inductionofcancerbyalternationsinseveraltypesofproteinsinvolvedincellgrowthcontrol,SignalTransductionandCancer,LectureII:IntracellularSignallingOutline:Whataresomesignallingpathways?Whataretheircellbiologicaloutputs?Howdotheseresultinthecancerphenotype?Howcanweexploitsignallingpathwaysfortherapy?,GenericSignalTransduction,RTKSignalTransduction,SignalTransductionDownstreameffectors,ProteinSignalingModules(Domains),SH2andPTBbindtotyrosinephosphorylatedsitesSH3andWWbindtoproline-richsequencesPDZdomainsbindtohydrophobicresiduesattheC-terminioftargetproteinsPHdomainsbindtodifferentphosphoinositidesFYVEdomainsspecificallybindtoPdtlns(3)P(phosphatidylinositol3-phosphate),MechanismsforActivationofSignalingProteinsbyRTKs,Activationbymembranetranslocation,Activationbyaconformationalchange,Activationbytyrosinephosphorylation,MechanismsforAttenuation&TerminationofRTKActivation,1)Ligandantagonists2)Receptorantagonists3)Phosphorylationanddephosphorylation4)Receptorendocytosis5)Receptordegradationbytheubiquitin-proteosomepathway,ActivationofMAPKPathwaysbyMultipleSignals,Growth,differentiation,inflammation,apoptosis-tumorigenesis,OverviewofMAPKSignalingPathways,TheMAPKPathwayActivatedbyRTK,P,RTKST-PI3Kpathway,Proto-oncogenesthatEncodeforSignallingProteins,Serine/ThreonineKinasesc-raffamilyaktNon-receptorTyrosineKinasessrcablReceptorassociatedbindingproteinsc-rasfamily,RasrecruitsRaftothemembrane,STintermediatescanbetargetsforanti-cancerdrugs,Kinases:Raf,STintermediatescanbetargetsforanti-cancerdrugs,Kinases:Bcr-Abl,CellPatterningCellGrowth,WntBMPHedgehogFGF,WhataretheessentialelementsofanySignalingcascade?,SignalligandDiffusibleorTetheredReceptorTransmembrane(exceptforlipidsolubleligands)Transducers-effectorsTargetsGenesorCellularcomponents,WntSignalingPathway,SignalWntsReceptorFrizzledsTransducers-effectorsb-cateninTargetsGenescytoskeleton,Wingless(Wg):Drosophilamorphogen-diff.Concentrationsofligandelicitdifferentresponsesinequivalentcellsmorphogenicmovementsandcellfatedeterminants“Beposterior”-cellfate“divide”-proliferationdevelopmentalabnormalitieswhengenedeleted,TheSignal:Wnt,Sharmadescribesawinglessmutationin1973,Sharma,1973Wingless-anewmutantinD.melanogaster.D.I.S.50:134SharmaandChopra,1976,Effectofthewingless(wg1)mutationonwingandhalteredevelopmentinDrosophilamelanogaster.Dev.Biol.48:461-465,Lateritwasclonedpositionally,IntegrationofMMTVcausesmammarytumorsinmice,TumorsarepregnancydependentMMTVhasasteroidenhancerMicedevelopbreasttumorsbutonlyduringlactationGenewasdesignated-Int-1(integrationofMMTV)OtherinsertionsitesoccurredatotherGFse.g.FGFTumorsexhibitdominantGainofFunctionLesson:Ectopicactivationofagenehyperplasia=Oncogene,Wingless+int-1=Wnt,FlywgandMouseInt-1arehomologs,Genesarecloned.Sequenceissimilar102030405060708090100HWnt-1MGLWALLPSWVSTTLLLALTALPAALAANS-SGR-WWGIVNIASSTNLLTDSKSLQLVLEPSLQLLSR-KQRRLIRQNPGILHSVSGGLQSAVFlyWgMDISYIFVICLMALCSGGSSLSQVEGKQKSGRGRGSMWWGIAKVGEPNNITP-IMYMDPAIHSTLRRKQRRLVRDNPGVLGALVKGANLAI110120130140150160170180190200HWnt-1RECKWQFRNRRWNCPT-APGPHLFGKIVNRGCRETAFIFAITSAGVTHSVARSCSEGSIESCTCDYRR-RGP-GGPDWHWGGCSDNIDFlyWgSECQHQFRNRRWNCSTRNFSRGKNLFGKIVDRGCRETSFIYAITSAAVTHSIARACSEGTIESCTCDYSHQSRSPQANHQAGSVAGVRDWEWGGCSDNIG210220230240250260270280290300HWnt-1FGRLFGREFVDSGEKGRDLRFLMNLHNNEAGRTTVFSEMRQECKCHGMSGSCTVRTCWMRLPTLRAVGDVLRDRFDGASRVLYGN-FlyWgFGFKFSREFVDTGERGRNLREKMNLHNNEAGRAHVQAEMRQECKCHGMSGSCTVKTCWMRLANFRVIGDNLKARFDGATRVQVTNSLRATNALAPVSPNA310320330340350360370380390400HWnt-1RGSN-RASR-AELLRLEPEDPAHKPPSPHDLVYFFlyWgAGSNSVGSNGLIIPQSGLVYGEEEERMLNDHMPDILLENSHPISKIHHPNMPSPNSLPQAGQRGGRNGRRQGRKHNRYHFQLNPHNPEHKPPGSKDLVYL410420430440450460470HWnt-1EKSPNFCTYSGRLGTAGTAGRACNSSSPALDGCELLCCGRGHRTRTQRVTERCNCTFHWCCHVSCRNCTHTRVLHECLNFlyWgEPSPSFCEKNLRQGILGTHGRQCNETSLGVDGCGLMCCGRGYRRDEVVVVERCACTFHWCCEVKCKLCRTKKVIYTCLN,TheSignal:Wnt,Secretedproteinligandsof80-100aaLipidmodifiedLatestbreakthrough(2003):purificationofactiveWntrequiresorganicextraction!ShortrangeactingSticktoextracellularmatrixGradients-morphogenic?multipleWnts(19inhuman/mouseand7inDrosophila),Wntssignalthroughserpentinereceptors,2classesofsignalingreceptorsCatalyticTyrosineKinaseReceptorsRTKsSer/ThrKinaseReceptorsBMPsSerpentine/G-protein-coupled-receptors(GPCRs)/7-transmembraneWntsadrenergic,dopamine,epinepherineetc,TheReceptor:Frizzled,corereceptorforWntsseven-passtransmembraneproteinsprobablyG-proteincoupledreceptorsmultipleFrizzleds(10inhuman/mouseand4inDrosophila),anewlyidentifiedco-receptorforWntssinglepasstransmembraneproteinrelatedtofamilyoflipoproteinreceptors,LRP/arrow:,WntSignalingregulatesgeneexpressionandcellpolarity,canonical,Lrp,non-canonical,CanonicalWntsignalingin2005,/rnusse/pathways/cell2/,/cgi/cm/CMP_5533,b-cateninisthecytoplasmic-nuclearsignalingmediator,b-catenin,b-catenin,armadilloinDrosophilageneticsdeterminedthatitfunctioneddownstreamofWgb-catenininmammaliansystemidentifiedascomponentofcelladhesionjunctionssubcellularlocalizationofproteincontroversialforyearspurificationofb-cateninandcloningofgenein1991byP.McCraeandB.Gumbinershowedthatarmadilloandb-cateninareorthologues,Thetransducer/effector:,Armadillorepeatstructureofb-catenin,CK1,E-cadherin,Wntsignalingpathway,C.Liuetal.2002.Cell108:837.,Complicatedphosphorylationcontrolsb-cateninstability,Howdoesb-cateninreachtargetgenes?,LEF/TCFtranscriptionfactorsHMG(HighMobilityGroup)proteinsmammalianLEF-1andTCF-1identifiedinTlymphocytesin1991twomoremembersclonedbylowstringencyscreeningofLibrariesanddegeneratePCRin1993b-cateninwasusedinayeasttwo-hybridassayandLEF-1wasclonedasaninteractingproteinin1997-endpointofthepathwaydetermined-mergedtwoindependentgroupsofscientists-subcellularlocalizationofb-cateninfinallysettled,GeneralStructureofLEF/TCFTranscriptionFactors,b-cateninbinding,Co-repressorbinding,DNAbinding/bending,alt.COOH,TCF-1,TCF-3,TCF-4,B,B,E,E,E,94%,96%,99%,55%,52%,64%,TargetGenesofWntSignaling,cellcycleregulatorsandtranscriptionfactors-c-MYC-cyclinD1tissuespecificgenestissueremodelingproteins-matrixmetalloproteinases-ephrinreceptorsandligands-adhesionproteinsangiogenesis-VEGF,IntheabsenceofWntsignaling:,Groucho,GSK-3b,APC,AdenomatousPolyposisColiIdentifiedbyJoannaGrodenandRayWhiteasatumorsuppressorgenesufferingLOHinfamilieswithveryhighratesofcoloncancer.TruncationmutationsorlossoftheentiregeneoccursinmostSporadiccoloncancers,/rnusse/wntwindow.htm,Hereditarycolorectalcancer(15%),FamilialAdenomatousPolyposis(FAP)-90%80%80%(prevalance)(germline)(somatic)(somatic),MMRdeficiency90%70%?65%mutations,APCshuttlemode-speculativel,Wntsignalingandcolorectalcancer,MajorfunctionofAPCistheregulationofcelluarb-cateninlevels.ActivationofwntpathwayincoloncancerdrivescellproliferationTcf-responsivegenes:c-myc,cyclinD1,PPARd-fibronectinandmatrilysin(anextracellularmetalloproteinase),CNS,Mutationclusterregion-allresultinproteintruncation,RacGEF,Graybars-b-cateninbindingsites.APCmayplayaroleincell-celladhesion(Cadherins)Redbars-Axin/Conductinbindingsites(lostinmutations)Redarrows-nuclearexportsignals.MutantAPCaccumulatesinthenucleusAsefbindingactivatesRacatmembranes,inducingmembranerufflingthereforepossiblyaffectingcellmotilityMT-microtubulebindingsite.APCisinvolvedinlinkingmicrotubulestokinetochoresthereforemutationscancontributetogenomicinstability,b-catenindestructioncomplex,AxinandAPCphysicallyinteract.APCmutationsincolonCAlackAxinbindingsites.-b-cateninbindstoAPC.APC/AxincomplexregulatesGSK3bkinaseactivity.BindstoAxin.ThereforeAxinmayserveascaffoldingfunction.AxinandAPCarealsoGSK3bsubstrates,andphosphorylationincreasestheirabilitytobindb-catenin.HowwntsignalsinhibitGSK3bactivityisunclear.Dishevellediscritical.WntsignalresultsindephosphorylationofAxinPP2AdephosphorylatesAxin.ItscatalyticsubunitbindsAxinwhileitsregulatorysubunitbindsAPC.TheregulatorysubunitofPP2AismutatedinasubsetofcolonCA.HowisPP2Aactivityregulated?-Whereistheintracellularlocalizationofthedestructioncomplex?,APCmutation,WildtypeAPC,APCmutationsresultinincreasedgenomicinstability,MouseModel-APCmin,Multipleintestinalneoplasia(min).APCgenemutation.Truncatedproteinatcodon850.Htzhaveincreasedpropensityfortumors.TumorsacquiresomaticmutationinwildtypeAPCallele.TumorslocatedinupperGItract(notcolorectal).Geneticbackgroundofmouseinfluencestumorload(?modifiers).MOM-1-possiblysecretedphospholipaseA2.APC1638TlacksC-terminaldomainthatbindstubulin,EB1-likeproteins.homozygousEScellshavehighdegreeofchromosomalinstabilitybuthomozygousmicedoNOTexhibitincreasedtumorsusceptibilityCooperatingOncogenes.Cyclooxygenase2:deletionofCOX-2genesuppressesintestinalpolyposisinAPCD716mice.COX-2levelsareincreasedinpremalignantpolyps.ButCOX-2isexpressedininterstitialcellsnotintestinalepithelium.Smad4:deletionofSmad4inAPCD716miceresultedinmoreaggressivetumors(compoundhtzmice).HighlightstheroleofTGFsignalintumorprogression.DNAmethyltransferase:compoundhtzhavereducedpolypnumbers(epigeneticevents?).,TumourProgression,TGFsignalingmutationsreceptorIImutationsdetectedinregionsofhighgradedysplasiabutabsentinadenomas.Intumourswithmicrosatelliteinstability(MI)mutationscorrelatewithprogressionofadenomastocancermutationsinTGFsignalingcomponents(e.g.,smad4)-nonMItumoraccelerate/worsenmurine(APCmin)intestinalcancermodel,Cell-celladhesivecomplexmutations-cadherins,b-catenins,others?,3.Metalloproteinaseactivation-matrilysinisatcf-responsivegene,compactionoftheearlyembryo,-morphogeneticmovementofcells-establishmentofcellfates,andpolarity,lossofcell-cellandcell-matrixrecognition,tissueinvasionmotility,normaldevelopment,cancerprogression,“epithelialmesenchymal”transition,HedgehogSignalingPathway,SignalHedgehogReceptorPatchedTransducers-effectorsCubitusInterruptusTargetsGenes,MutationsinHedgehogsignalinginhumansembryosyieldscyclopia(aformofholoprosencephaly)imagesareonlyforthestout-hearted.Inadults,mutationsinHedgehogsignalinggivesphenotypesinstemcellandprogenitorpopulations.Increasedsignalinggivestumors,lesssignalinggivesshort-livedstemcellsMostrecentadvanceisthatmanytumorsshowelevatedHhsignaling.Cyclopamine(firstobtainedfromthecornlily)haspromisefortherapeuticinterventionofcancer.,TheSignal:Hedgehog,Secretedproteinligand-heavilyprocessedLipidmodifiedwithcholesterolShortrangeactingSticktoextracellularmatrixGradients-morphogenic?threeligandsinmammals:Indian,Desert,Sonic,Thehedgehoggeneencodesanovelmembrane-linkedligandimportantforlocalpatterningofmanytissues.Theprimarytranslationproductcontainsasignalpeptidethatiscleavedtoproducea45kDapolypeptideprecursor.Cleavageofthissecretedprecursorproducesa20kDaN-terminalfragmentassociatedwiththeplasmamembraneandwithinductiveactivityplusa25kDafragment.TheN-terminalfragmentbecomestetheredtothemembraneviaahydrophobiccholesterolmoiety(itdoesntcontainanyhydrophobicresidues).Aseriesofelegantexperimentsshowthatthisarrangementensuresthathedgehogonlyactsontheadjacentcellanddoesntdiffusetoactoncellsfurtheraway(i.e.itisspatiallyrestricted).,TheSignal:Hedgehog,Hedgehogligandismodifiedbycholesterol,TherearefewerHedg