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1、1889 1889 种子与土壤假说种子与土壤假说种子与土壤假说种子与土壤假说1909 1909 免疫监视免疫监视免疫监视免疫监视1910 1910 病毒与肿瘤病毒与肿瘤病毒与肿瘤病毒与肿瘤1915 1915 激素与癌症激素与癌症激素与癌症激素与癌症1937 1937 肿瘤干细胞肿瘤干细胞肿瘤干细胞肿瘤干细胞1939 1939 血管再生术血管再生术血管再生术血管再生术1950 1950 吸烟与肿瘤吸烟与肿瘤吸烟与肿瘤吸烟与肿瘤1953 1953 二次突变假说二次突变假说二次突变假说二次突变假说1960 1960 染色体易位染色体易位染色体易位染色体易位1971 1971 肿瘤抑制基因肿瘤抑制基因
2、肿瘤抑制基因肿瘤抑制基因 细胞凋亡与肿瘤细胞凋亡与肿瘤细胞凋亡与肿瘤细胞凋亡与肿瘤1975 1975 肿瘤微环境肿瘤微环境肿瘤微环境肿瘤微环境19761976克隆演变克隆演变克隆演变克隆演变&多步骤肿瘤发生多步骤肿瘤发生多步骤肿瘤发生多步骤肿瘤发生 19761976病毒癌基因的细胞同系物病毒癌基因的细胞同系物病毒癌基因的细胞同系物病毒癌基因的细胞同系物 1978 1978 癌基因编码癌基因编码癌基因编码癌基因编码 蛋白调控细胞生长蛋白调控细胞生长蛋白调控细胞生长蛋白调控细胞生长 1979 1979 第一个人类癌基因第一个人类癌基因第一个人类癌基因第一个人类癌基因 1983 1983 癌基因互作
3、癌基因互作癌基因互作癌基因互作 、肿瘤表观遗传学、肿瘤表观遗传学、肿瘤表观遗传学、肿瘤表观遗传学 1989 1989 细胞周期和细胞周期和细胞周期和细胞周期和DNADNA损伤检查损伤检查损伤检查损伤检查1990 1990 肿瘤易感性的遗传基础肿瘤易感性的遗传基础肿瘤易感性的遗传基础肿瘤易感性的遗传基础 肿瘤遗传不稳定性的机制肿瘤遗传不稳定性的机制肿瘤遗传不稳定性的机制肿瘤遗传不稳定性的机制 1999 1999 肿瘤谱肿瘤谱肿瘤谱肿瘤谱 2001 2001 肿瘤靶向治疗肿瘤靶向治疗肿瘤靶向治疗肿瘤靶向治疗 肿瘤研究百年大事记肿瘤研究百年大事记（Nature Milestones in Cance
4、r, milestones of timelines)1889 1889 种子与土壤假说种子与土壤假说种子与土壤假说种子与土壤假说 (Seed and Soil Hypothesis) (Seed and Soil Hypothesis) Stephen Paget (assistant surgeon to the West London hospital)By analyzing 735 case histories of fatal breast cancer, he found that metastasis formed in the liver far moreeven those
5、 such as the spleen that could be considered to have the same exposure to the cancer cells because of similar blood flows.（Nature, milestones of timelines)Question: “what is it that decides what organ shall suffer a case of disseminated cancer”Conclusion: Cancer cells require some “nourishment nouri
6、shment” from the environment to develop.Molecular mechanism (until today)?1975 1975 肿瘤微环境肿瘤微环境肿瘤微环境肿瘤微环境 (Tumor Microenvironment) (Tumor Microenvironment) Beatricemintz and Karl IllmenseeThey took the tetracarcinoma cells from embryoid bodies in vivo, and injected them into developing mouse blastocy
7、sts. Surprisingly, normal mice were born with no evidence of tumors. They found that tumor-derived cells were present in large numbers and contributed to several unrelated tissues, the most notable being functional spermatozoa. They concluded tumor cells were developmental totipotent and could rever
8、t to normal behavior in the appropriate environment. （Nature, milestones of timelines)1984 1984 肿瘤微环境肿瘤微环境肿瘤微环境肿瘤微环境 (Tumor Microenvironment) (Tumor Microenvironment) Mina BissellContext is everythingPeople and idea, JCB, 2009The tumor-inducing behavior of Rous sarcoma virus (RSV) when injected into
9、 the wings of newly hatched chicks was already known, and the viral gene v-src had been identified as the sole culprit. What Bissell found was that if RSV was injected into 4-day-old embryo, no tumor were produced, despite the spread of RSV infection and active v-src expression. Other contributions
10、:The wounding is important for tumorigenesisBlocking integrin function is sufficient to revert the maglignant phenotype of human breast cancer.General concept of tumor microenvironmentTumor microenvironment refers to the unique properties of tissue microenvironment conferred by abnormal interactions
11、 between tumor and host cells. Tumor microenvironment is often characterized by hypoxia, nutrient deprivation, acidosis, and aberrant stroma. Highly variable, with differences seen between patients and often in different areas of the same tumor. The tumor microenvironment is often altered as the dis
12、eases progresses; even the percentage of a tumor made up of cancer cells may change. 基本定义基本定义:主要特征主要特征:Tumor microenvironmentTumor cellTumor-associated stromal cells （fibroblast，endothelial cell, Immune cell）Extracellular Matrix Proteins （structural and non-structural)AcidicCytokinesNeurotransmitter
13、s HypoxiaGrowth factorsMMPs, ADAMs, other secreted proteins and their receptors HormoneChemokinesTumor-associated stromal cellsActivated fibroblast ( hepatic stellate cell)Immune cellEndothelial cellPericyteAdipocyteGlial cellTumor associated fibroblastCharacteristics of normal fibroblastActivated f
14、ibroblast in the tumor microenvironmentOrigin of activated fibroblasts in the tumor stromaInteraction between tumor cells and activated fibroblastsThe role of activated fibroblast in cancer developmentCross-talk between activated fibroblasts and other tumor-associated host cellCharacteristics of nor
15、mal fibroblastBasic component of connective tissue and contribute to structural integrityMain resource of ECM and regulation of ECM turnoverLow proliferate index and minimum metabolic capacityMain resource of growth factors and cytokinesActivated fibroblast in the tumor microenvironmentSimilar pheno
16、type as fibroblasts in inflammation and wound healingIndented nuclei, stress fibers and well-developed cell-matrix interactionMarkers: Fibroblast activation protein (FAP), a-smooth muscle actin (SMA), r-smooth muscle actin (gSMA), Prolyl-4-hydroxylase (P4H, for collagen synthesis)Identification: -SM
17、A in combination with at least 3 other markers (e.g., positive for -SMA, vimentin and P4H and negative for cytokeratin)基本特征：基本特征：如何鉴定：如何鉴定：Origin of activated fibroblasts in the tumor stromaSurrounding resting fibroblasts after their activation and recruitment to the tumor tissue, Bone marrow stem c
18、ell, pericytes, via epithelial or endothelial to mesenchymal transition.Xouri, et .al. 2010, seminars in cell and developmental biologyInteraction between tumor cells and activated fibroblastsTumorActivated fibroblastsTGF-b, PDGF, LL-37, LPAAll the stages of tumor developmentThe role of activated fi
19、broblast in cancer developmentInitiation of tumorKuperwasser, et al., 2004, PNASBhnowmick, et al., 2004, ScienceProgression of tumorBhnowmick, et al., 2004, ScienceMetastasis of tumorGrugan , et al., 2010, PNASDuda, et al., 2010, PNASTGF-beta signaling in fibroblasts modulates the oncogenic potentia
20、l of adjacent epithelia.Bhowmick NA, Chytil A, Plieth D, Gorska AE, Dumont N, Shappell S, Washington MK, Neilson EG, Moses HL.Science. 2004 Feb 6;303(5659):848-51.Conditional knock-out TGF-beta type II receptorFibroblastsAbundance of stromal cellsIntraepithelial neoplasia in prostate （5-7 weeks）Inva
21、sive squamous cell carcinoma of the forestomach （7-week-old）Paracrine HGF signaling（FSP1 promotor）3 weeksInitiation of tumorFibroblast-secreted hepatocyte growth factor plays a functional role in esophageal squamous cell carcinoma invasion.Grugan KD, Miller CG, Yao Y, Michaylira CZ, Ohashi S, Klein-
22、Szanto AJ, Diehl JA, Herlyn M, Han M, Nakagawa H, Rustgi AK.Proc Natl Acad Sci U S A. 2010 Jun 15;107(24):11026-31. Epub 2010 Jun 1.HGF/MET signalingInvasion of tumorMalignant cells facilitate lung metastasis by bringing their own soilDuda DG, Duyverman AM, Kohno M, Snuderl M, Steller EJ, Fukumura D
23、, Jain RK Proc Natl Acad Sci U S A. 2010 Dec 14;107(50):21677-82 Metastasis of tumorCross-talk between myofibroblasts and other tumor-associated host cellWever et al. 2008, Int.J.CancerSuppression of antitumor immunity by stromal cells expressing fibroblast activation protein-alpha.Kraman M, Bambrou
24、gh PJ, Arnold JN, Roberts EW, Magiera L, Jones JO, Gopinathan A, Tuveson DA, Fearon DT.Science. 2010 Nov 5;330(6005):827-30.Transgenic mouseFAP (fibroblast activation protein-) ) promotorExpressing Primate diphtheria toxin receptorEndothelial cells and tumorTumor angiogenesis: Therapeutic implicatio
25、n. N. Engl. J. Med.1971 Judah Folkman In 1939, Gordon Ide and colleagues In 1945, Glenn Algire and colleagues In 1968, Melvin Greenblatt and Philippe Shubik He proposed that tumour cells secrete a soluble factor that stimulates the proliferation of endothelial cells, that these in turn control tumou
26、r expansion and, in the absence of new vessel growth, that tumours do not increase beyond 23 mm in size, entering instead a state of dormancy. Stromal endothelial cells directly influence cancer progression.Franses JW, Baker AB, Chitalia VC, Edelman ER.Sci Transl Med. 2011 Jan 19;3(66):66ra5Secretio
27、ns from quiescent endothelial cell(EC-conditioned medium)Breast and lung carcinoma proliferation and invasivenessKnockdown of perlecanEliminates EC ability to suppress cancer invasivenessInhibitTumor microenvironmentTumor cellTumor-associated stromal cells （fibroblast，endothelial cell, Immune cell）E
28、xtracellular Matrix Proteins （structural and non-structural)PHCytokinesNeurotransmitters ？HypoxiaGrowth factorsMMPs, ADAMs, other secreted proteins and their receptors Extracelluar Matrix The extracellular matrix (ECM) the extracellular part of animal tissue that usually provides structural support
29、to the animal cells in addition to performing various other important functions. The extracellular matrix is the defining feature of connective tissue in animals.Extracellular matrix includes the interstitial matrix and the basement membrane.Interstitial matrix: present between various animal cellsB
30、asement membrane: sheet-like depositions of ECM on which various epithelial cells restComposition of ECMProteinsCollagensLaminins-G-X-Y-, b, gsubunitFibronectinElastinsProteoglycans (蛋白聚糖）蛋白聚糖）Syndecan (硫酸乙酰肝素蛋白聚糖蛋白聚糖)1. Classical extracellular matrix 2. Matricellular proteins Osteopontin (OPN), Ost
31、eonectin (SPARC), thrombospondins, tenascin, CCNsBiology of CollagensStructure (Triple helix)Basic Composition-Glycine-Proline-Hydroxyproline (hydroxylysine)-Types of Collagen29 types of collagens have been foundCollagens is the most abundant protein in mammals, making up about 25% to 35% of the who
32、le-body protein content UsageCosmetic, MedicalLaminins:ubiquitous proteins of basement membranesEpitheliaVasculatureMuscleGlandNerveLaminins: a family of proteins111, 121211, 221, 213511, 521, 5231, 2, 5 1, 3 1, 23A4 1, 3 1, 2311, 321411, 421, 4233A 2 3332A3B 2 3332BSpecific spatio-temporal expressi
33、on 111embryonic 211, 221mainly muscles411, 421mainly endothelium332mainly epithelium511, 512more ubiquitousAumailley et al.,A simplified laminin nomenclatureMatrix Biol., 2005, 24, 326-3325 , 3 , 3 subunits, 16 lamininsSubunits of lamininsLAMA1: 3075 amino acids, 337084 Da; 17 EGF-like domain, 5 Lam
34、G domain; 1 LamNT domain, 2 LamB domain. subunitLAMB1: 1786 amino acids, 198066 Da ; 13 EGF-like domain, 1 LamNT domain, 1 PDB domain (Protein Data Bank, chemotaxis). subunitLAMC1: 1609 amino acids, 177603 Da ; 10 EGF-like domain, 1 LamNT domain, 1 LamB domain, 1 Scop domain (Structural Classificati
35、on of Proteins, Apolipophorin-III). subunitRoles of ECM in Biology Providing support and anchorage for cells Segregating tissues from one another Regulating intercellular communication Reservoir of growth factorsECM and ECM fragments can function as growth factorECM in pathological processesWound he
36、alingFibrosisCancer BiologyInvasionMetastasisTumor ProgressionInflammationExtracelluar Matrix A Matter of Life and Death Anoikis is a form of programmed cell death which is induced by anchorage-dependent cells detaching from the surrounding extracellular matrix (ECM).Anoikis and cancerMetastatic tum
37、or cellsEscape from anoikis and invade other organs Cancer stem cellExtracelluar Matrix A Matter of Life and Death Pro SURVIVAL ECMFibronectin prosurival2386 amino acids; 262625 Da FNI domain, FNII domain, FNIII domain15 FNIII domain, RGD motif for integrins and other protein interactionConfer resis
38、tance to apoptosis; activates ERK1/2, ROCK and PI3K/AKT pathwayPro SURVIVAL ECMFibronectin prosurivalFibronectin 5 1 integrinFAKERKRhoACell survivalPro SURVIVAL ECMRescues cells from apoptosis by protein kinase B/AKT activationLaminin 10/11 5 1 1, 5 2 1Laminin 10/11 3 1 integrinp130casPI3K/AKTRac1Ce
39、ll survivalExtracelluar Matrix A Matter of Life and Death ECM stiffness (Physical properties)Leads to integrin clusterization and triggers mitogenic signalsPhysico-mechanical aspects of extracellular matrix influences on tumorigenic behaviors.Cukierman E, Bassi DE. Semin Cancer Biol. 2010 Jun;20(3):
40、139-45. ECM stiffness influence tumor initiation, progression and metastasisFrom fibrosis to cancerFriends or foeRoles of ECM in cancerFriends or foeMatrix crosslinking forces tumor progression by enhancing integrin signalingLevental et al., 2009, Cell This article offer a new paradigm for understan
41、ding why tumor incidence so dramatically with aging.ECM stiffness on celluar behaviorsExtracelluar Matrix A Matter of Life and Death Pro APOPTOTIC ECMCCN1Induces apoptosis through a6b1 integrin and syndecan-4 bindingCCN family (6 members):1.cysteine-rich protein 61 (Cyr61 or CCN1)2. connective tissu
42、e growth factor (CTGF or CCN2)3. nephroblastoma (肾母细胞瘤 ) overexpressed protein (Nov or CCN3)4.Wnt-inducible secreted protein-1 (WISP-1 or CCN4)5. WISP-2 (CCN5) 6.WISP-3 (CCN6)SPARCInduces apoptosis through a yet unknown mechanismDecorinInduces apoptosis through activation of caspase-3 by an unknown
43、mechanismEMILIN2Induces apoptosis through death receptors DR4 and DR5 direct bindingPro APOPTOTIC ECMEndostatinInduces apoptosis through downregulation of apoptosis inhibitorsEMILIN2 (Elastin Microfibril Interface Located Protein 2)Matricellular ProteinsAt the crossroad of infalmmation and cancerMat
44、ricellular proteins are a class of non-structural ECM protein exerting regulative functions likely through a direct binding to specific surface receptors, to growth factors and cytokines, to other matrix proteins.Matricellular ProteinsAt the crossroad of infalmmation and cancerOPN (Osteopontin)Wound
45、 healingAffected phases during healingHealing rateMatrix deposition and contractionAcceleratedInflammationRole in inflammationLeukocyte functionPro-inflammatory Th1 committingPro-survival of lymphocytes, monocytes and neutrophilsTumorigenesisRole in tumor-host interactionEffect on tumor growthPro-an
46、giogenicInvolved in tumor progression and angiogenesis. Level of expression correlated with stages of disease (breast and prostate)Matricellular ProteinsAt the crossroad of infalmmation and cancerTSP-1Wound healingAffected phases during healingHealing rateTGFbactivationinflammationIn transgenic mice
47、 overexpression TSP-1InflammationRole in inflammationLeukocyte functionResolving inflammationRecruits monocyte and neutrophilsNegative regulator of DC and Th1 effectors activation neutrophils chearanceTumorigenesisRole in tumor-host interactionEffect on tumor growthStromal TSP-1 is anti-angiogenicTu
48、mor growth, Angiogenesis and activates TGFbHealing in also in TSP-1 KO miceMatricellular ProteinsAt the crossroad of infalmmation and cancerTSP-2Wound healingAffected phases during healingHealing rateCollagen depositionAcceleratedInflammationRole in inflammationLeukocyte functionInflammation and ang
49、iogenesis in TSP-2 KO miceTumorigenesisRole in tumor-host interactionEffect on tumor growthStromal TSP-2 is anti-angiogenicTSP-2 deficiency susceptibility to skin carcinogenesis andtumor formationRoles of ECM in cancerFriends or foeDesmoplasia Appears at relatively early-stage lesions and disappears
50、 in highly invasive, advanced tumor It could be a host defense reaction designed to confine the developing tumor by tumor associated with stromal cells Fragments of ECM in AngiogenesisRoles of ECM in AngiogenesisCollagen XVIIIinhibits angiogenesisC-terminal fragment, EndostatinTrimeric NC1 domainpro
51、motes angiogenesis1 NC domain, 1 FRI (Frizzled ) domain, 1 LamG, 6 Collagen doamains, 1 EndostatinRoles of Endostatin in CancerPerlecanPromote tumor growth angiogenesisendorepellinC-terminal fragment from domain V of Perlecaninhibits angiogenesisRoles of ECM in AngiogenesisRoles of ECM in cancer inv
52、asion and metastasisRoles of ECM in cancer invasion and metastasisLaminin 332 Laminin 332 2 chain has been found at leading edge of invading carcinomasBladder, Breast, Cervical, Colorectal, Gastric, Head and neck, Liver, Lung, Pancreas, prostate and many othersOsteopontinVersicanPrognostic factor, p
53、roliferation, adhesion, motility,metastasis, angiogenesisPromotion of metastasis, inhibition of apoptosis, neoangiogenesisCarcinoma-produced factors activate myeloid cells through TLR2 to stimulate metastasis.Kim S, Takahashi H, Lin WW, Descargues P, Grivennikov S, Kim Y, Luo JL, Karin M.Nature. 200
54、9 Jan 1;457(7225):102-6.VersicanTN-a， IL-6MetastasisECM as a circulating marker for cancer diagnosisTumor MarkerMain Application Field1. Mucins, proteoglycans, glycoproteinsEpisialin and related (MUC-1 family, CA15-3, CA 549, CAM29, CAM26, CASA, BCM, KL-6, MCA, TAG72)Sialyl Lewis and related (CA 19-
55、9, GICA, CA 195)Cancer antigen 125 and related related (CA125, CA602, OVX)TRA-1-60Soluble glypican-3Soluble mesothelinCarcino-embryonic antigen and related (CEA, NCA)2. Host-derived mediatorsOsteoprotegerin, bone alkaline phosphatase3. Soluble ECM breakdwon productsLaminin fragmentsCollagen type IVT
56、ype I Cllagen Carboxyterminal Telopeptide (ICTP)Circulating ECM complexes (DR-70)Breast carcinomaPancreatic, bile duct and colorectal carcinomaOvarian carcinomaTesticular germ cell tumor ( no-seminous)MelanomaMesotheliomaColorectal carcinomaSkeletal metastasis (prostatic cancer)Colorectal carcinomaB
57、reast carcinomaSkeletal metastasis (prostatic cancer)Lung carcinomasheet-like depositions of ECM on which various epithelial cells restBasement membraneBMBasement membrane structureUltrastructureKeratinocyteBasementmembraneKeratinsBP230PlectinCD151CollagenXVIICollagenIVInterstitialcollagensCollagenV
58、IILM332 6 4CellmembraneDermis3070 nanometers in thicknessBMMMPsBMBMInvasion through the basement membraneCellproliferation+increasingintra-tumoralpressure= disruptionof BMTumor cellextendsfilopodiaandmigratetowardsdermalsignalDiffusiblemediatorDermalalterationThe receptor of ECMIntegrin familyNon-in
59、tegrin family18 subunit, 8 subunit, more than 20 integrinsSyndecan, -dystroglycan, 67 kD laminin receptor (Laminin receptors)DDRs, GPVI, LAIR-1, uPARAP/Endo180 (collagen receptors)Integrin activation4 subfamily integrins47E5683b10125891473611Collagen-binding integrinLaminin-bindingintegrinsRGD-bindi
60、ngintegrinsL2MXDLeukocyteintegrinsECM receptors- integrin familyRoles of Integrins in cancerRegulation of cell survival and apoptosisRoles of Integrins in cancerRegulation of angiogenesisRoles of Integrins in cancerRegulation of migration and invasionRoles of Integrins in cancerRegulation of cancer
61、stem cellIntegrins interaction with receptors of growth factorVEGF Receptor (Ruoslahti 2002)TGFB Receptor (Scaffidi et al. 2004)c-MET (HGF receptor) (Sridhar & Miranti 2006)PDGF Receptor (Sridhar & Miranti 2006)EGF Receptor (Bill et al. 2006)Type-1 insulin-like growth factor receptor (Bill et al. 20
62、06)Insulin Receptor (Schneller et al. 1997)Integringrowth factor and integrincytokine receptor crosstalkIntegrins in the host defense to cancerIntegrins as targets for cancer therapyRoles of MMP in cancerMatrix metalloproteinases (MMPs) are zinc-dependent endopeptidasesCapable of degrading all kinds
63、 of extracellular matrix proteins, but also can process a number of bioactive moleculesInvolved in the cleavage of cell surface receptors, the release of apoptotic ligands (such as the FAS ligand), and chemokine/cytokine in/activationFunctionsProliferationMigrationDifferentiationAngiogenesisApoptosi
64、sHost defenseRoles of MMP in cancer(A family of zinc-dependent endopeptidases)MMPs have been implicated in Cancer for more than 40 yearsMMP-mediated ECM degradation leads to cancer cell invasion and metastasis has been a guiding pricipleNew insights into the Roles of MMPs in CancerMMPs affect growth
65、 signalsMMPs regulate apoptosisRegulation of tumor vasculatureInitiation of Neoplastic progressionTissue invasion and metastasisMetastatic niche formationOrchestrate inflammation in cancerNew insights into the Roles of MMPs in CancerMatrix metalloproteinases: regulators of the tumor microenvironment
66、.Kessenbrock K, Plaks V, Werb Z. Cell. 2010 Apr 2;141(1):52-67. ReviewMMPs and tumor microenvironmentOther proteinases in ECM turnoverADAMs (A Disintegrin And Metalloproteases) inflammation, cancer, Alzheimers particularly in tumor-stroma interactionTumor microenvironmentTumor cellTumor-associated s
67、tromal cells （fibroblast，endothelial cell, Immune cell）Extracellular Matrix Proteins （structural and non-structural)AcidicCytokinesNeurotransmitters ？HypoxiaGrowth factorsMMPs, ADAMs, other secreted proteins and their receptors Metastasis and angiogenesis induced by hypoxiaThe process of metastasis
68、often occurs in regions of hypoxia, in which HIF can induce expression of pro-invasive and pro-angiogenic factors, such as LOX, CTGF, VEGF,Ang-2 and decreases in E-cadherin and thrombospondin. Tumor acidic microenvironment OxygenNutrientDeprivationAbnormal metabolism (Warburg effect)GlycolysisLactic
69、 acidacidic microenvironmentChemoresistanceNormal cell deathECM degradation and cancer metastasis德国生物化学家, 曾获1931年诺贝尔生理学-医学奖 Neurotransmitters and cancerSystem regulation of cancer microenvironmentBMECMIntegrinsCytokinesActivated fibroblastImmune cellChemokinesNeurotransmitterEpithelial cellCarcinoma Growth factor神经系统神经系统免疫系统免疫系统内分泌系统内分泌系统Together !We can beat cancer !
