1、癌症分子生物学癌症分子生物学(研究生研究生)全册配套完整全册配套完整课课件件Class 1The Nature of CancerTumor, like normal tissue, are compose of mass of cells, but are unable to assemble and create tissue of normal form and function Cancer, is a disease of malfunctioning cellsWhat is a tumor or cancer ?The Biology of Cancer 2007肿瘤产生于正常组
2、织肿瘤产生于正常组织normal mucosa (正常粘膜正常粘膜)dysplastic mucosa (异常粘膜异常粘膜)adinocarcinoma (腺癌腺癌)小肠小肠(回肠回肠)内壁切片显示了正常组织与癌组织之间的延续性内壁切片显示了正常组织与癌组织之间的延续性The Biology of Cancer 2007Stroma(mixture of fibroblasts, adipocytes and collagen)Stroma(基质基质)Milk ductBreast adenoma与正常组织一样，肿瘤也是有细胞组成与正常组织一样，肿瘤也是有细胞组成正常乳腺切片正常乳腺切片 乳腺
3、癌组织乳腺癌组织切片切片The Biology of Cancer 2007StromaStroma (基质基质)Milk ductBreast carcinoma肿瘤能侵入邻近正常组织肿瘤能侵入邻近正常组织正常乳腺切片正常乳腺切片 乳腺乳腺癌组织切片癌组织切片The Biology of Cancer 2007B16鼠黑色素瘤细胞尾静脉注射鼠黑色素瘤细胞尾静脉注射2周后周后在在WT 和和 g g-/- 小鼠肺的黑色素瘤转移灶小鼠肺的黑色素瘤转移灶Metastases -new settlements beyond original tumor sitesWT g g -/- 肿瘤能转移到其他
4、正常组织形成转移灶肿瘤能转移到其他正常组织形成转移灶大肠癌的肝转移大肠癌的肝转移 ( (白色部分白色部分) )乳腺癌的脑转移乳腺癌的脑转移 Primary tumor (原发瘤原发瘤):tumor at original tumor sitesMetastasis (转移瘤转移瘤,转移灶转移灶):new settlements beyond original tumor sitesBenign tumor(良性瘤良性瘤):without invading adjacent tissuesMalignant tumor(恶性瘤恶性瘤): invaded nearby tissues and sp
5、awned metastasesAdenoma (腺瘤腺瘤):premalignant epithelia growthCarcinoma (癌癌):malignant epithelia growthIn fact, the great majority of tumor in human are benign and harmless,except for : 1. localized mess to press on vital organ 2. thyroid adenomas hyperthyroidism( 甲状腺功能亢进症甲状腺功能亢进症) 3. pituitary adenom
6、as acromegaly (肢端肥大症肢端肥大症）Metastasis are responsible for some 90% of deaths from cancer常见与肿瘤相关术语或名词常见与肿瘤相关术语或名词 I. Epithelial cells: carcinoma(癌） squamous cell carcinoma adenocarcinomaII. Nonepithelial cells: 1. Connective tissues: sarcoma(肉瘤, from mesenchymal cells)2. Hematopoietic tissue： leukemia
7、，lymphoma3. Nervous system: neuroectodermal tumors(神经上皮瘤)III. Some types of tumors not fitting the major classification:Small cell lung carcinomas (SCLCs)肿瘤由体内多种特殊化细胞产生肿瘤由体内多种特殊化细胞产生Adenomas or carciomas:Readily be detected with naked eyes，dysplastic under microscopeNot penetrated base membraneNeopl
8、asia( (瘤生瘤生) ): both benign and malignant tumorMetastases: form new clones in remote sitesMultiple steps in Cancer developmentMetaplasia: 正常正常细细胞被原来不在同一区域的另一种胞被原来不在同一区域的另一种细细胞取代，胞取代， an early indication of premalignant change( (组织变形组织变形) )Hyperplasia: 细细胞数量增加，但与正常胞数量增加，但与正常组织组织相比相比细细胞学上只有些微差异胞学上只有些微
9、差异( (增生增生) )Dysplasia: 细细胞学异常、胞学异常、细细胞形胞形态态异常异常variability of nuclear size & shape increased ratio of nuclear versus cytoplasmic sizeIncrease mitotic activitylack of cytoplasmic features of differentiated cellscytological changes:( (异常异常) )MetastasesHyperplasia( (增生增生) )Dysplasia ( (异常异常) )Neoplasia
10、( ( 瘤生瘤生) )Normal( ( 转移转移) )Normal micro-structure of epithelium a collecting tube of the kidneyThe columnar epithelium of the gallbladderthe bronchiole of the lungThe endometrium of the uterusThe Biology of Cancer 2007The Biology of Cancer 2007鸡角膜表皮扫描电镜显微照片Normal ultrastructure of epithelium小鼠气管表皮切
11、片透射电镜显微照片HyperplasiaMammary epithelial cells have begun to pileup and to protrude into lumina. More advanced hyperplastic mammary duct show epithelial cells almost completely fill the lumen, but they have not penetrated the basement membrane yet.The Biology of Cancer 2007StromaMilk ductMetaplasiaThe
12、 Biology of Cancer 2007The squamous cells in esophagus is replaced by secretory cells migrated from stomach. To increase 38 folds for malignancy.Figure 2.14 The Biology of Cancer ( Garland Science 2007)DysplasiaThe Biology of Cancer 2007Dysplasia is a transitional state between complete benign growt
13、h and premalignantThe Biology of Cancer 2007Adenoma, or polyps papilomas or carcinoma, ( or wart in skin)Breast carcinomasColonic adenomasThe Biology of Cancer 2007Epithelial cells:squamous cells squamous cell carcinoma (鳞状细胞癌鳞状细胞癌) secretory cells adenocarcinoma (腺癌腺癌)responsible for responsible fo
14、r 80%80% of the cancer-related death in western world of the cancer-related death in western worldThe Biology of Cancer 2007Squamous cells (鳞状细胞鳞状细胞)/ )/Squamous cells carcinomas (鳞状细胞癌鳞状细胞癌) )宫颈上皮宫颈上皮皮肤上皮皮肤上皮正在侵入基质的食管癌正在侵入基质的食管癌胃壁上形成小囊，排列整胃壁上形成小囊，排列整齐的粘液分泌细胞齐的粘液分泌细胞小肠壁上单个粘液分泌细胞小肠壁上单个粘液分泌细胞Secretory
15、 epithelia / adenocarcinomasAdenocarcinomas in stomach （胃腺癌）胃腺癌）Adenocarcinomas in colon （大肠腺癌）大肠腺癌）Epithelial cells:carcinomasquamous cell carcinomaadenocarcinomaNonepithelial cells:1. Connective tissues: sarcoma (from mesenchymal cells); 1%2. Hematopoietic tissues: 3. Nervous system:80%肿瘤由体内多种特殊化细
16、胞产生肿瘤由体内多种特殊化细胞产生Leiomyosarcoma(平滑肌瘤)sarcomas derived from mesenchymal cells in connective tissuesOsteosarcoma (骨瘤)Liposarcoma (脂肪瘤)The Biology of Cancer 2007Tumors arise from many specialized cell types through the bodyEpithelial cells:carcinomasquamous cell carcinomaadenocarcinomaNonepithelial cel
17、ls:1. Connective tissues: sarcoma (from mesenchymal cells); 1%2. Hematopoietic tissues: leukemia lymphoma:T and B lymphocytes aggregated solid mess in lymph nodeerythocytesantibody-secreting cellsT and B lymphocytes3. Nervous system:80%Common hematopoietic malignancyALLAMLCLLCMLMMALLAMLCMLMMerythrol
18、eukemiaThe Biology of Cancer 2007Tumors arise from many specialized cell types through the bodyEpithelial cells:carcinomasquamous cell carcinomaadenocarcinomaNonepithelial cells:1. Connective tissues: sarcoma (from mesenchymal cells); 1%2. Hematopoietic tissues: leukemia lymphoma:T and B lymphocytes
19、 aggregated solid mess in lymph nodeerythocytesantibody-secreting cellsT and B lymphocytes3. Nervous system: neuroecodermal tumors80%2.5%The Biology of Cancer 2007Ganglia cellsGranular layer(GL)正常神正常神经组织经组织各种神各种神经经瘤瘤Some types of tumors do not fit into the classification1. melanomas2. small- cell lu
20、ng carcinomas3. epithelian-mesenchymal transition (EMT)4. anaplastic( (退行发育退行发育) ) or dedifferenciation( (去分化去分化) )Melanocytes and melanomasMelanocytes are pigmented cells of the skin and retina, Developed from neurocrest, but no directed connection with nervous systemThe Biology of Cancer 2007Melan
21、ocytes in skinpigment granules in a melanocyte viewed by EMMelanoma metastasesSmall cell lung carcinomas (SCLCs)Contain cells having many attributions of neurosecretory cellsResponse to neuronal signalingSecrete biologically active peptidesFrequently seen in tobacco usersNot clear about the origin o
22、f the SCLCs Epithelial-mesenchymal transitionEpithelia cancer cells at the border of many carcinomas often change shape and gene expression programs and take on attributes of nearby stromal cells of mesenchymal origin. Anaplastic( (退行发育退行发育) ) tumorsThe Biology of Cancer 2007In the above three situa
23、tion, cancer cells almost always retain some of the distinctive attributes as critical clues for pathologists to define the origin of cancer cells. BUTMultiple steps in Cancer developmentMetastasesHyperplasia( (增生增生) )Dysplasia ( (异常异常) )Neoplasia( ( 瘤生瘤生) )Normal( ( 转移转移) )The Biology of Cancer 200
24、7monoclonality versus polyclonality of TumorsThe Biology of Cancer 2007Tumors are monoclonal growthIH for G6PD of intestine sectionsThe Biology of Cancer 2007肿肿瘤的瘤的单单克隆起源克隆起源The Biology of Cancer 2007 肿肿瘤瘤单单克隆起源的其他克隆起源的其他证证据据骨髓瘤骨髓瘤细细胞只胞只产产生一种抗体生一种抗体The Biology of Cancer 2007 肿肿瘤瘤单单克隆起源的其他克隆起源的其他证证据据
25、不同人种的不同人种的肿肿瘤瘤发发生率是生率是same or different ？每个成年人个体平均每个成年人个体平均细细胞数：胞数：1013每个成年人一世平均每个成年人一世平均细细胞数：胞数：1016每个人每秒每个人每秒钟钟死亡或被替死亡或被替换换的的细细胞数：胞数：107Some are same, same different depending on tumor types.Some caused by random, unavoidable accidents of nature occur at similar frequencies in various human popula
26、tions, eg for certain pediatric tumors.The Biology of Cancer 2007Tumor incidence is different in different human populationThe Biology of Cancer 2007Environments affect tumor incidenceLife style affects tumor incidenceThe Biology of Cancer 2007The Biology of Cancer 20071915年，年，Dr. Katsusaburo Yamagi
27、wa 用煤焦油首次在兔耳成功用煤焦油首次在兔耳成功诱导肿诱导肿瘤瘤The Biology of Cancer 2007The Biology of Cancer 2007Structure of carcinogenic hydrocarbonsFigure 2.23a The Biology of Cancer ( Garland Science 2007)Figure 2.23b The Biology of Cancer ( Garland Science 2007)Figure 2.24 The Biology of Cancer ( Garland Science 2007)Figu
28、re 2.25 The Biology of Cancer ( Garland Science 2007)Table 2.8 The Biology of Cancer ( Garland Science 2007)leiomyosarcomasarcomas derived from mesenchymal cells in connective tissuesAdipocytes: liposarcomaOsteoblasts: osteosarcomaMyocytes: leiomyosarcomaFibroblasts: fibrosarcomaosteosarcomaLiposarc
29、omaFigure 2.16a The Biology of Cancer ( Garland Science 2007)Figure 2.16b The Biology of Cancer ( Garland Science 2007)Invasive carcinomaTumor are considered malignant only after they have penetrated base membrane and invaded into nearby surronding stroma.Breast ductal carcinomThe Biology of CancerC
30、lass 2Tumor Viruses肿瘤病毒在人类癌症研究中的地位肿瘤病毒在人类癌症研究中的地位病毒导致人类许多人类疾病( 如狂犬病， 天花，感冒等）绝大部分病毒在细胞内疯狂复制增殖,杀死细胞而引起疾病某些病毒的增殖不杀死细胞，还导致细胞不受控制地疯狂生长 肿瘤病毒诱发肿瘤的病毒。1970s肿瘤病毒研究证实：人类肿瘤一小部分是肿瘤病毒诱发的。肿瘤病毒研究提供了揭开隐藏数百年的人类肿瘤秘密的钥匙癌症是一种基因病。可用分子生物学和遗传学进行分析研究肿瘤病毒革命了人类癌症病理学的研究Peyton Rous Peyton Rous 肿瘤病毒学的奠基人肿瘤病毒学的奠基人1910年，发现劳氏肉瘤病毒(R
31、ous sarcoma virus, RSV)引发鸡肿瘤50多年后(1966) 获得Nobel Prize in Medicine and PhysiologyThe Biology of Cancer, 20071876年，一位俄国科学家发现通过组织移植肿瘤在不同的狗之间转移， 提出肿瘤也是一种传染病的假说1908年，两位哥本哈根科学家从鸡白血病细胞制备出一种可过滤的鸡白血病致病原Rous sarcoma virus (RSV)Rous sarcoma virus (RSV) induces chicken tumorsThe Biology of Cancer, 2007提供了一种随意诱导
32、肿瘤产生的研究肿瘤的工具， 肿瘤研究不必依赖不可预期的动物或人类自发肿瘤的出现。A setback in research on tumor viruses1913年丹麦人Johannes Grib Fibiger报道了大鼠胃中的螺旋菌导致胃瘤 1926年 Johannes Grib Fibiger 获得了Nobel Prize所称胃瘤实际上是一些组织变形的胃表皮(meteplastic stomach epithelia). 组织变形的原因是严重缺乏维生素丢掉了Fibiger的实验动物丢弃了感染性致病因子会引发肿瘤的学说相差光镜相差光镜 照片照片扫描电镜扫描电镜 照片照片正常正常CEFRSV
33、转化的转化的CEFRSVRSV转化的转化的CEFCEF细胞失去了接触抑制特性细胞失去了接触抑制特性 在长满细胞的培养皿中形成在长满细胞的培养皿中形成focifociThe Biology of Cancer, 20071950代后期加州理工学院的博后Hurry Rubin和博士生Howard Termin发现： 1. RSV能感染培养皿中的鸡胚成纤微(CEF)细胞, 并复制病毒 2. RSV感染的CEF不裂解，能无限 生长, 即RSV能转化CEF RSVRSV能在培养皿中转化能在培养皿中转化( (transformtransform) ) CEF CEF细胞细胞RSV能使正常细胞转化成肿瘤细胞
34、。The Biology of Cancer, 2007Renato Dulbecco Howard Temin1975 1975 的医学和生理学的医学和生理学Nobel PrizeNobel PrizeAn RSV-induced focus肿瘤病毒研究的再生及意义肿瘤病毒研究的再生及意义意义：肿瘤的发生可以从个体细胞水平进行研究。意义：肿瘤的发生可以从个体细胞水平进行研究。 他们的发现再次改变了他们的发现再次改变了2020世纪的癌症研究进程。世纪的癌症研究进程。Questions:是是RSV病毒感染、转化了病毒感染、转化了the progenitor cell of focus, 并且并且
35、RSV病毒继续存在与子代细胞中维持转化状态病毒继续存在与子代细胞中维持转化状态？或:RSV病毒以病毒以 “hit and run” 的方式最初作用于的方式最初作用于progenitor cell, 造成转化细胞的一系列形态和生理变造成转化细胞的一系列形态和生理变化，然后化，然后RSV病毒从犯罪现场消失了病毒从犯罪现场消失了？维持细胞的转化状态依赖于维持细胞的转化状态依赖于RSVRSV存在和活性存在和活性The Biology of Cancer, 200719701970年，年， University of California Berkeley University of Californi
36、a Berkeley分校发分校发现了一株温度敏感突变型的现了一株温度敏感突变型的 RSV RSV病毒病毒Anchorage independent growth RSVRSV转化转化CEFCEF细胞细胞在软琼脂中形成克隆在软琼脂中形成克隆and tumorigenecityThe Biology of Cancer, 2007在在athymiathymic c nude micenude mice形成肿瘤形成肿瘤转化细胞的特征转化细胞的特征细胞形态的改变细胞形态的改变(圆形圆形, 在相差显微镜下有折射性在相差显微镜下有折射性)；丢失接触抑制丢失接触抑制(contact inhibition)特
37、征特征 (ability togrow over on another)，能高密度生长；，能高密度生长； 无需附着固体生长的能力无需附着固体生长的能力(anchorage independence growth)；无限生长的能力无限生长的能力 (immotalization)；减少对生长因子需求或依赖性；减少对生长因子需求或依赖性；增加了葡萄糖运输能力；增加了葡萄糖运输能力；具有成瘤性。具有成瘤性。1910Rous sarcoma virus (RSV)50s-60sMurine mammary tumor virus(MMTV) Human T-cell leukemia virus (HT
38、LV-1) Avian leukosis virus (ALV) 1930s Papillomavirus (to induce papilloma ( (乳头乳头( (状状) )瘤瘤) ) in rats） ）50s-60s Hepatitis B virus (HBV， ，肝炎病毒肝炎病毒) SV40 ( (能在猴细胞中复制增殖能在猴细胞中复制增殖, , 但只能转化鼠细胞但只能转化鼠细胞） polyomavirus ( (多瘤病毒多瘤病毒，polyoma in mice） Human papilloma 16 (HPV) Human adenovirus 5 ( (人上呼吸道感染，诱导仓鼠
39、肿瘤）人上呼吸道感染，诱导仓鼠肿瘤） DNA viruses:RNA viruses:RNA RNA 和和 DNA DNA病毒都能诱导肿瘤病毒都能诱导肿瘤Harald zur HausenThe Nobel Prize in 2008 （发现人乳头瘤病毒诱发宫颈癌）Papovavirus Papovavirus 的的DNADNA基因组的电镜照片基因组的电镜照片一个一个RSVRSV相关病毒相关病毒的的RNARNA基因组电镜照片基因组电镜照片(A)(A)和示意图和示意图(B)(B)RNARNA病毒和病毒和DNA DNA 病毒含有不同的遗传物质病毒含有不同的遗传物质RSVRSV病毒颗粒含有病毒颗粒含
40、有两份两份RNARNA线性线性基因组基因组大部分大部分DNADNA病毒颗粒含有病毒颗粒含有一份一份DNADNA环状环状基因组基因组DNA viruses:RNA viruses:宿主DNA聚合酶; RNA聚合酶; 核糖体自身编码以RNA为模板的 RNA聚合酶;宿主核糖体自身编码以RNA为模板的 DNA聚合酶;宿主核糖体 RNA RNA病毒颗粒和病毒颗粒和DNADNA病毒颗粒结构不同病毒颗粒结构不同The Biology of Cancer, 2007RSVRSV病毒病毒Shope papillomavirus电镜电镜照片照片 Shopepapillomavirus冰冻冰冻蚀刻电镜照片蚀刻电镜照
41、片根据根据X光衍射推出的光衍射推出的 SV40 病毒结构病毒结构DNADNA病毒病毒Enter host cellsMultiplyQuickly kill their hosts, Release progeny virus particles from dying cellsRe-infect other susceptible cells in the vicinity.Simple life cycle for most of DNA virusesThe Biology of Cancer, 2007RSVRSV和其他和其他RNARNA病毒以出芽的方式从细胞释放病毒以出芽的方式从细胞
42、释放The Biology of Cancer, 2007Murine lekemia virus病毒感染细胞的扫描电镜照片病毒感染细胞的扫描电镜照片 The Biology of Cancer, 2007DNA病毒诱导的细胞转化也依赖于DNA病毒的全部或部分基因组在细胞表达Large TLarge T是是 SV40 DNA SV40 DNA病毒的一个肿瘤相关抗原病毒的一个肿瘤相关抗原SV40 DNASV40 DNA病毒诱导的转化细胞的细胞核呈现病毒诱导的转化细胞的细胞核呈现Large TLarge T抗原阳性抗原阳性The Biology of CancerTransmitted via v
43、irus infection?New SV40 viruses are only made during lytic cycle of SV40Transformed cells are nonpermissive for producing new virusesTransmitted via cell division ?viral DNAs replicate as autonomous, extrachromosomal mol.Nonpermissiveness prevents the repl. of SV40 virual DNALimited number of SV40 D
44、NA molecules in the transformed cells.Question: How does SV40 DNA virus genome in a transformed cell transmit over many cell generation? Integration of SV40 DNA into chromosomal DNA DNA molecules from SV40-transformed cells and SV40 DNA molecules from free SV40 virus were co-centrifuged in alkaline-
45、sucrose gradient. The fractionations collected were then analyzed by UV-spectrophotometer and DNA hybridization. The Biology of Cancer99.7%99.7%人宫颈癌细胞携带人人宫颈癌细胞携带人HPVHPV病毒基因组含致癌基因的病毒基因组含致癌基因的DNADNA片片断，而不是整个断，而不是整个HPVHPV病毒基因组。病毒基因组。 Human cervical carcinoma: Figure 3.6 The Biology of Cancer ( Garland
46、Science 2007)Puzzle about the stable transmission of RSV genome重复感染假说重复感染假说1. 1.某些某些RSV病毒突变株能感染和转化细胞，病毒突变株能感染和转化细胞， 但不能在被转化的细胞中复制但不能在被转化的细胞中复制2 2 . . 某些细胞能被转化，但不能被再感染某些细胞能被转化，但不能被再感染。Howard Temin3 3 . . 抑制抑制DNA复制或复制或 DNA-dependent RNA 合成可抑制合成可抑制RSV病毒的复制。病毒的复制。Any Question ?The Biology of Cancer, 200
47、7Howard TeminProvirus Theory(The life cycle of Retrovirus)19701970年，年， Howard Termin Howard Termin 和和 David David Baltimore Baltimore 发现反转录酶发现反转录酶1975 1975 的医学和生理学的医学和生理学Nobel PrizeNobel PrizeThe Biology of Cancer, 2007Important distinction between integration mechanisms used by retrovirus and DNA v
48、irusRetrovirus: normal & essential part of replication cycle whole genome integrated at most time DNA virus: rare accident (80%鸡白血病鸡白血病ALV原病毒都整合在原病毒都整合在c-myc座位座位1.ALV proviruses integrated at 5 of c-myc in 80% of chicken leukemia2.Most integrated in the same transcriptional orientation as that of c-
49、mycIn 1981The Biology of Cancer, 2007慢转化反转录病毒通过插入诱变激活原癌基因慢转化反转录病毒通过插入诱变激活原癌基因The Biology of Cancer, 2007Examples of cellular genes found to be activated by insertional mutagenesis(Wnt-1)(FGF)(notch receptor)insertional mutagenesis can be used as a strategy to screen for new proto-oncogenes Some retr
50、oviruses naturally carry oncogenes1.HTLV-1, belong to the third class of retroviruses, naturally carrying oncogenes;2.infects 1% of the inhabitants of a south island of Japan, and some islands of the Caribbean, 3-4% risk of developing adult T-cell leukemia;3.proviruses are integrated randomly;4.carr
