《resources肿瘤问题PPT文档资料.ppt》由会员分享，可在线阅读，更多相关《resources肿瘤问题PPT文档资料.ppt（42页珍藏版）》请在三一办公上搜索。
1、1,1.现有“生物信息学”的限制：“有眼不识泰山”第一外显子 识别能力 外显子内含子 识别能力2.一个基因：不同剪切体 Splicing form(Isoform)3.一个基因 蛋白质 切割：蛋白质 1 肽段,人基因组计划已经给了我们多少？,人基因有多少个？,10万 3万？,2,4.同一个蛋白质 细胞内的不同“定位”不同功能！5.98 DNA序列：功能不明!（95）,细胞核,细胞质,线粒体,细胞膜,蛋白质移位 Translocation,3,E1 b E1 a Exon 2 Exon 3,p16 Ink 4a,P14 ARF(human)P19 ARF(mouse),Cross Talk be
2、tweren Oncogenes&tumor suppressors,Oncogenic SignalsRas,c-myc,E2F,p16 Ink 4a,Rb,on,Cell cyclearrest,P14 ARF(Block MDM-2/p53 binding),p53 on,DNADamage,一个基因可编编码两种完全不同的蛋白质,4,功能基因组学 Functional Genomics,阐明基因（蛋白质）之间的相互作用 与生命活动的关系 与疾病的关系 与环境、药物的关系基因组测序只是基因组研究的起点“功能基因组学”的含义被“局限地”误解“基因表达谱”的误解或局限性 cDNA芯片：mRNA
3、表达蛋白质表达 蛋白质芯片：只知道蛋白质的表达与存在 并不知道蛋白质的功能状态,问题：,在基因组的水平,5,1.糖基化(1)膜表面蛋白：影响细胞识别，细胞细胞相互关系(2)细胞表面抗原：(3)细胞内蛋白：特定糖基化与蛋白质叠成为高级结构有关 成熟蛋白质不成熟蛋白质的“质控”表志2.乙酰化：组蛋白：与DNA结合能力、复制、转录 其它蛋白质（如p53）3.磷酸化：4.蛋白质切割成片段：一个蛋白质多种功能。,6,细胞分子细胞生物学重要问题,过度增殖：恶性肿瘤 高血压（内皮细胞、平滑肌细胞）细胞凋亡：神经退行行病变 早老性痴呆 帕金森氏病“To be or not to be,thats a ques
4、tion”HamletShakespear,细胞增殖,细胞凋亡,生 死,7,（抗凋亡）Antiapoptotic,（促进凋亡）,M,Check Points of Cell Cycle G1 S check point G2 M check point M:Mitotic Spindle checkpoint,8,分子 细胞 组织 机体,误 区：相互分离。每种疾病均集中于反映出疾病的细胞肿 瘤：集中在癌细胞 误 区：忽视癌细胞以外的细胞心血管疾病：心脏病 心肌细胞 心肌间质血管内皮细胞 高血压 内皮细胞 内皮细胞间质细胞炎症细胞 平滑肌细胞 神经性疾病：神经原细胞 神经原 胶质细胞 间质 内皮
5、细胞（高血压脑病）,9,间质细胞血管内皮细胞淋巴管内皮细胞淋巴细胞巨噬细胞触突状细胞 Dendritic cell,Cytokine(细胞因子)Cytokine Receptor 细胞因子受体Chemokine(趋化因子)Chemokine Receptor趋化因子受体,恶性肿瘤,除肿瘤细胞外，还有重要的细胞：,10,CANCER:,Multiple Cross-talk among Cancer,stromal and endothelial Cells,Cancer cells,Stromal cells,Endothelialcells,Intercellular Cross talk,
6、?,Cancercells,Matrix(ECM)Endothelial cellsPericytesStromal cells,11,Stroma:,1.Fibroblasts in tumor:“Activated fibroblast”GF GF R activated 2.Genetic alteration:Juvenile polyposis syndrome(JPS),Cowen syndrome etc Dominant hereditary Stromal cells abnormality,Hamartoma High risk for colon cancer PTEN
7、and others“Landscaper”“Stroma abnormal microenviroment”“Microenviroment of tumor-host interface”,12,ANGIOGENESIS,Two Processes Endothelial Cells(EC)growth migrationVessel formation Tube formation extensionOther cells:Fibroblasts etc,13,ANGIOGENESIS/CANCER INVASION,EC Growth control&Vessel formation,
8、Hypoxia Activating hypoxia responses genes(p53)Acidosis VEGF-A(Cancer.EC),14,Protease/matrix/Cancer cell invasion/Angiogenesis,Proteases:Degradation of ECM and Cell Associated Proteins MMPs(Marix metallo proteinases),membrane-anchored MMP 2,MMP 9,(Cancer Cell)Soluble MMP(fibro,EC)Adamalysin related
9、membrane proteinases BMP 1 metalloproteinases Tissue serine proteinases tPA UK Thrombin Plasmin uPA(EC,fibro.)uPAR(Cancer),Inhibitors PAI TIMPs(Tissue inhibitor of MMPs)PEX(Degradation product of ECM by MMP 2),15,Proteases further create Plasminogen Angiostatin(38KD)EC ATP synthase(I)Procollagen XXI
10、II Endostatin(20KD)Prolaction 16KD prolactinECM PEX EC integrin ab3(I)RGD fragment Cancer FAK(S)Troponin Tn-1 EC ATP Synthase(I)(Human cartilage),?,Inhibitors of AngiogenesisStimulators of Cancer Invasion,(I),ANGIOGENESIS/CANCER INVASION,16,EC Receptors:(EC and Cancer Cell)VEGF R 1(Flt-1)VEGF R 2(Fl
11、k-1,KDR)VEGF A-D PLGF VEGF R 3(Flt-4)VEGF C,D TIE 1?TIE 2 Ang 1,2 3?4?,*Integrin ab3*ECM PEX ab5(Laminin,tenasin)CD 36 Thrombospondin(ECM Glycoprotein)cell adhesion,motility&growth metallospondin,VascularTubuleformation,*Eph B1 Recognizing some ligands or partners NHE 3?ARNT?EPA-1?,Tenasoin:new ECM.
12、,ANGIOGENESIS/CANCER INVASION,17,VEGF Rs,VEGF R1 VEGF R2 VEGF R3 Neurophilin 1 Neurophilin 2(flt-1)(flk-1 KDR)(flt 4),Ligands:,18,EC Growth Factors:,Specific GFs:VEGF A-D VEGF R2(KDR.Flk-1)PLGF VEGF R1(flt-1),Non-specific GFs(Local):Some of them:autocrine(EC,Cancer Fibro,)TGFa EGF R TGFb(fibro)TGFb
13、R(Cancer)TGF b/ECM TGF b released aFGF bFGF FGF R PDGF PDGF R HGF Met,ANGIOGENESIS/CANCER INVASION,Protease,19,Chemokines,CXC chemokines Recruit monocytes(Tumor cell origin)leukocytes or othersCXC L12 CXC L-12 enriched tissueCCR4 metastasisPseudopodia,migration,penetration of ECM,homing,Induce TNFa,
14、Cytokines related to EC or Capillary formation,IFN a Downregulate VEGF production bFGFIFNg Inhibit angiogenesisIL-12 Upregulate IFN-g&IP-10,inhibiting angiogenesis,ANGIOGENESIS/CANCER INVASION,20,Interaction between cancer cells&EC,HypoxiaProteases Degradation of ECMVEGFsAngsTGEabFGF PDGF?Chemokines
15、,cancer,EC,ANGIOGENESIS/CANCER INVASION,Metastasis,21,Enigmas,1.Tumor/endothelial cells Cross-talk2.Tumor cells endothelia like cells.Morphological evidence:yes Biochemical:?3.Stromal cells/tumor cells Stromal cells/endothelial cells4.Genetic alteration of tumor vascular endothelial cells?Yes or not
16、?5.Genetic alteration of tumor stromal cells,Cross-talk,22,APOPTOSIS,1.Death signals:TNFa family2.Death Receptors:TNF Receptor family,Decoy Receptor3.Adaptor molecules:Interact with receptor Caspase(Protocaspase)4.Proto caspases:Activated Caspases5.Effector molecules:Other Proteases,DNA degradation
17、enzymes6.Inhibitors:For Caspases Adaptors Others,New Progress,Molecules inside mitochondria release into cytoplasm(Procaspases,cytochrome C ctc.)Nuclear/Cytoplamic molecules translocate to MT membrane,23,Mitochondrion/Cytoplasm,Caspases Other Signals,In intermembrane spaceProtocaspase 2 9 3Latent AI
18、FCytochrcme CSmac/DIABLO,Life,Other targest,Mitochondrion,Protocaspase-2Protocaspase-3Protocaspase-9Apaf 1Cytochrome cAIFOthers?(Smac/DIABLO),Life or Death?,Alteration of MolecularCompartmentation(Mitochondrion versus Cytoplasm),Other Targets,Intermembrane Space,AIF(Apoptosis inducing factor)Science
19、,397:3879 1999Smac(Second MT-derived activator of caspase)Or DIABLO(Direct IAP-binding protein at Low pI),Bcl 2 guard?,Death,24,Mitochondria-death Signal Integrators,C.Bremer&G.Koemer(Science.Aug.18.289:1150,2000),Translocate to mitochondrinStimulation/inductionInhibition/block translocation(Keep in
20、 cytosol),PTPC:Permeability Transition Pore Complex(Bax etc.)Bcl2/Bax/Bid permeabilize the outer MT membrane upon interaction with PTPC Bid may be independentJNK:(SAPK,stress activating protein kinase)-inactivate Bcl XL p53:Translocated to MT&interacted with hsp 70 PKC:Ttranslocated to MT,25,Enigmas
21、 in Apoptosis,Translocation of nuclear or cytosolic protein molecules into mitochondria,Phosphorylation/dephosphorylationTranslocator Protein/protein bindingconformational changeBinding to mitochondria surface membane protein,Translocation or release of proteins or other molecules from mitochordria
22、to cytoplasm,Permeability transit pore complex Voltage depedent anion channel(VDAC),Apoptosis induced by paradoxical signals,Growth factor/receptorsProtooncogenes:Ras,c-myc etc.,26,HOT SPOTS OF SIGNALING,20%of 32,000 human coding gene:Signal transductionProtein Kinase 520Protein Phosphatase 130Domin
23、ant Oncogenes 100 PKTumor Suppressor 30 PKKnown PTKPTK 90(May,2001)RPTK 58(20 families)Non-receptor,PTK 32(10 families)Cytoplasmic,27,Human RPTK,28,c-src:Truncated mutation of C to Tyr 530 autophosphorylation related to STAT-3 in colon cancerc-abl:CS 9(9:22)abl/BCR in ph1 CS DNA damage induced apopt
24、osis.ATM c-abl bind Rb at G0/G1(released after RB phosph.of abl prevent DNA-damage induced abl phosph.PDGF Induce cell motility&adhesion(c-abl involved in)BCR/abl Cyteplasm X nucleus,Human Cytoplasmic PTK,29,1.Class I p110(catalytic)Adaptor/regulator IA:RPTK activated IB:hetero-trimeric G protein co
25、upled Receptor activated.Class II p1(3)k2.Substrate:ptd Ins(4.5)P2 ptd Ins(3.4.5)p3 Some P1(3k)(I&III):+Ser/thr K 3.Isoform a,b,d of p110.Subunit p85a,p85b,p55g of adaptor4.Binding of protein to lipid IA FVVE domain ptd Ins(3)P PH(pleckstrin homology)domain PH(+)protein:3-phospho-inositide dependent
26、 K(PDK 1)(ser/thr PK)Akt(PKB),PI(3)K/Akt&mTOR/p7056k(rapamycin target),PI(3k):,hetero dimer,IA,ptd Ins(3,4)P2 ptd Ins(3,4,5)P3,30,Akt(PKB),Akt:v-Akt homolog PKB a,b,g isoforms N PH domain Central kinase domain activation loop:Thr.308 C Regulatory site:Ser 473PDK-1(3-phospho-inositide dependent Kinas
27、e 1):Thr 308 Akt kinase C PH domain 10 fold binding affinity to lipid membrane(constitutively binding)PDK-2:Ser 473 Akt,31,Substrate of Akt-1,32,Substrate of Akt:13:RXRXXS/T motif(this motif:also for MAPKAPK-1&p7056K),Two major class,33,RPTK activation P1(3)k Ptd Ins(3,4,5)P3 Ptd Ins(3,4)P2 Interact
28、 with AKT Akt/PDK1 Akt translocatin to membrane,Thr 308(loop)Ser 473(Reg.Site)(PDK2?),PDK1/Akt activation:,Akt,translocate to nuclei,AKT,PDK1 also PKC isoforms,serum and glucoorticoid induced K(GSK)PKC related K(PRK)p7056k p21 activated PK(PAK),34,PTEN,LOF of PTEN Akt also p27LOF in glioblastoma,ger
29、m cell cancer,breast cancerCancer&Pl(3)K IA/Akt Akt expression:pancreatic,ovarian cancer Akt/p65(mutant of p85 a Pl(3)k)p110(Pl(3)K)amplification(ovarian cancer)Pl(3)K IB(p110 g)LOF:colon cancer(Bcl2,CDK.Cyclin D),(3-phosphoinositide phosphatase,TS)ptd Ins(3,4,5)P3 ptd(3,4)P2,35,mTOR(Mammalian Targe
30、t of Rapamycin),1.Conserved family:Yeast:TOR1,TOR2,MEC1,TEL1,Rad 3 Drosophila:MEC41 Mammal:mTOR,ATM,ATR(ATM related)TRAPP(transformation/transcriptional domain-associated P)2.mTOR=FK506 binding P(FKBP)(human)Rapamycin associated P(FRAP human)Rapamycin and FKBP12 target-1(rat)3.mTOR C terminal K doma
31、in(homologous to kinase domain of Pl(3)K and Pl(4)K ser/the K activity only mTOR c-myc(inhibited by mTOR inhibitor rapamycin)stat 3(ser 727)PKC;PKC 4E-BP dissociation&release of elf-4E cap dependent initiation translation of 5-UTR,36,P1(3)K dependent cyelin D3 Phosphorylation of Rb and p107 E2FInvol
32、ved in P1(3)K and Akt tumorigenesisDependent on Phosphorylation&activation of p70S6K,Ribosomal S6 Kinase S6K1,S6K2,Regulator of cell growth&protein translationp70S6K short isoform of S6K 1 Cytoplasic in large part Activation is blocked by rapamycin Mediate mTOR effect on protein-translation Phosphor
33、ylation of S6 subunit(40s rs protein)S6 translation of 5 ter.Pyr-rich tract(5-TOP mRNA),Also involved in cell cycle,37,Wnt Pathway,Wnt Wnt 1=(int-1,mouse MMTV integration activated gene)Porcupine:A gene for wnt secretion Polytopic membrane protein Related to acyl transferaseFrizzled(Fz)Wnt Receptor
34、Dally:Co-receptor of Wnt Glypican-type heparan sulfate proteoglycan Glycosyl-Ptd Ins moiety membrane b-catenin:in absence of Wnt Destabilized by a protein complex Dishevelled(Dsh)activated by Wnt-Fz binding Axin APC GSK-3b(Glycogen synthase kinase)GSK-3b-b-catenin-b catenin degradation Wnt Fz bindin
35、g Dsh GSK-3b b-catenin stabilized enter nucleusNuclear transcription factor:Tcf/LEF:activated by b-catenin,38,Wnt Pathway,TCF/LEF,APC,GSK 3b,TCF/LEF,b-catenin,LRP,Fz,39,Hedgehog Pothway,Hedgehog:Embryonic development inducer Hh,hh:Humna Shh(Sonic hh)Ihh(Indian hh)Dhh(Desert hh)19KD protein C Cholest
36、erol ester N Palmitoylation(after cholesterol addition,enhancing signaling)Secretion of hh:2 genes:Dispatched related to hh R Patched hh release Toutvelu:enzyme,heparin-SO4 synthesis for transport and response of hhRececeptor of hh Patched(Ptc):Ts,12 Span-transmembrane Proton-driven lipid transporto
37、r Smoothened(Smo):Onc,7-transmembrane p(Fz like),40,1.hh(-):(in a complex)hh(+):2.Smo activated:coupled with a complex3.hh(-):(1)Ci N ter.transcription repressor(Ci 75)(2)Ci in cytoplasm(Cos2/Fu/Su/Ci complex anchored on microtubule)4.hh(+):Costa/Fu/Su/Ci dissociated from microtubule Ci(Full length)
38、:enters nucleus Transcription of genes(ptc,Hip and growth genes),Activation of hh Pathway,hh Ptc Smo*,hh-Ptc,Smo*,Smo*,Fu:Fused,S/T PKSu:Suppressor of FuCosta 2(Cos.2):Kinase likeCi:Cubitus interuptus,ZF TA Gli in mammal,PKA,41,Su,Fused(Fu),Costa 2,Smoothened(Smo),Patched(Prc),Hedgehog(Hh),C i,C i,Growthgeneproduct,PtcHip,Patehed:Disrfunctin in BCC Smo:mutated in breast Hedgehog Pathway,MiTmicrotubule,Mit,42,
