1、u物质代谢及酶的变化物质代谢及酶的变化u肿瘤细胞的分化肿瘤细胞的分化u肿瘤细胞的生长肿瘤细胞的生长u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因核酸代谢核酸代谢核酸增多是肿瘤迅速生长的物质基础核酸增多是肿瘤迅速生长的物质基础DNADNA拓扑异构拓扑异构酶酶u物质代谢及酶的变化物质代谢及酶的变化端粒酶端粒酶DNADNA拓扑异构拓扑异构酶酶存在于细胞核内的一类酶，他们能够催化存在于细胞核内的一类酶，他们能够催化DNADNA链的链的断裂和结合，从而控制断裂和结合，从而控制DNADNA的拓扑状态。的拓扑状态。DNADNA拓扑异构酶通过形成短暂的单链裂解拓扑
2、异构酶通过形成短暂的单链裂解-结合循环，结合循环，催化催化DNADNA复制复制的拓扑异构状态的变化的拓扑异构状态的变化 核酸代谢核酸代谢u物质代谢及酶的变化物质代谢及酶的变化(A)Classification of human DNA topoisomerases.Type IB are the only enzymes that form(A)Classification of human DNA topoisomerases.Type IB are the only enzymes that form cleavage complexes(cc)with 30-phosphotyrosyl
3、(30-P-Y)intermediates.cleavage complexes(cc)with 30-phosphotyrosyl(30-P-Y)intermediates.(C)Noncovalent binding of type IB enzymes.(D)Scheme of the 30 phosphotyrosine(C)Noncovalent binding of type IB enzymes.(D)Scheme of the 30 phosphotyrosine covalent bond in the Top1cc.The arrow indicates the rever
4、sible(religation)reaction,which is covalent bond in the Top1cc.The arrow indicates the reversible(religation)reaction,which is favored under normal conditions.G)Scheme of the 50-phosphotyrosine covalent bond in the favored under normal conditions.G)Scheme of the 50-phosphotyrosine covalent bond in t
5、he Top2cc.Top2cc.It is thought that length or integrity of chromosome It is thought that length or integrity of chromosome end is used as a mitotic counting mechanism end is used as a mitotic counting mechanism in vitro in vitro 核酸代谢核酸代谢u物质代谢及酶的变化物质代谢及酶的变化端粒酶端粒酶Each mammalian chromosome end has a di
6、stinctive DNA-Each mammalian chromosome end has a distinctive DNA-protein structure,which prevents the degradation and fusion protein structure,which prevents the degradation and fusion of chromosome ends by helping distinguish chromosome of chromosome ends by helping distinguish chromosome ends fro
7、m a double strand break in the genomic DNA.ends from a double strand break in the genomic DNA.Mammalian have a stretch of a simple repeat sequence unit(TTAGGG)and in ,length is 1520 kb.Fifty to 200 bp of the telomeric DNA shortens at each round of mitosis.When average DNA reaches a critically short
8、length,about 47 kb,is arrested Irreversibly.核酸代谢核酸代谢端粒酶端粒酶u物质代谢及酶的变化物质代谢及酶的变化u物质代谢及酶的变化物质代谢及酶的变化核酸代谢核酸代谢蛋白质代谢蛋白质代谢糖代谢糖代谢酶系统酶系统u物质代谢及酶的变化物质代谢及酶的变化酶系统酶系统增殖相关和分化相关的酶增殖相关和分化相关的酶转化相关和演进相关的酶转化相关和演进相关的酶细胞恶变的指标。细胞恶变的指标。主要正常细胞发生转化，主要正常细胞发生转化，总可出现这类酶活性的总可出现这类酶活性的改变。改变。演进相关的酶演进相关的酶 酶活性于恶性程度呈平行关系的酶酶活性于恶性程度呈平行关系
9、的酶转化相关转化相关u肿瘤细胞的分化肿瘤细胞的分化各种不同各种不同类型细胞类型细胞(分化细胞分化细胞)同一来源的同一来源的幼稚细胞幼稚细胞?分化的概念分化的概念特定的生理功能特定的生理功能特定的生化特征特定的生化特征特定的形态结构特定的形态结构稳定性稳定性全能性全能性选择性选择性条件可逆性条件可逆性细胞分化特点细胞分化特点:未分化恶性肿瘤是由于起源组织中的干细胞未分化恶性肿瘤是由于起源组织中的干细胞丧失了分化的能力。丧失了分化的能力。肿瘤细胞分化异常的机制肿瘤细胞分化异常的机制遗传学改变遗传学改变信号转化异常信号转化异常微环境的影响微环境的影响诱导分化治疗肿瘤诱导分化治疗肿瘤u肿瘤细胞的生长肿
10、瘤细胞的生长细胞增殖活性的原位检测方法及意义细胞增殖活性的原位检测方法及意义细胞增殖活性：细胞增殖活性：细胞增生快慢的能力细胞增生快慢的能力DNA DNA 含量测定含量测定 确定增生细胞的比例确定增生细胞的比例DNA ploidy and proliferative activity as represented by DNA ploidy and proliferative activity as represented by the S-phase fraction(SPF).the S-phase fraction(SPF).A link exists between high SPF
11、values and increased A link exists between high SPF values and increased risk of recurrence and death for patients with primary risk of recurrence and death for patients with primary BC,BC,Flow cytometry Flow cytometry 法法u肿瘤细胞的生长肿瘤细胞的生长免疫组织化学方法免疫组织化学方法Several monoclonal antibodies reacting with diff
12、erent Several monoclonal antibodies reacting with different proliferating cell nuclear antigens have been described,such proliferating cell nuclear antigens have been described,such as PCNA,Ki-67 and MIB 1,KiS1 and others.as PCNA,Ki-67 and MIB 1,KiS1 and others.The Ki-67/MIB 1 protein has a The Ki-6
13、7/MIB 1 protein has a prognostic valueprognostic value for many for many types of malignant tumors.types of malignant tumors.Ki-67Ki-67Only papers published in English in peer reviewed journals before June 2004 that include at least 100 evaluable patients were selected.Inaddition,the prognostic and
14、predictive role of the proliferative markers had to be assessed through multivariate analyses.One hundred and thirty-two papers fulfilled these criteria and 159 516 patients analyzed.u肿瘤细胞的生长肿瘤细胞的生长免疫组织化学方法免疫组织化学方法细胞周期蛋白细胞周期蛋白The different cyclinsThe different cyclins：the concentration rise and fall
15、 at the concentration rise and fall at specific stages throughout the cell cycle,have a specific stages throughout the cell cycle,have a temporally temporally distinctdistinct and and highly regulated pattern of expressionhighly regulated pattern of expression,i.e.they are,i.e.they are synthesized a
16、nd degraded at specific stages of the cell cycle.synthesized and degraded at specific stages of the cell cycle.Cyclin E is the limiting factor for G1 phase progression and Cyclin E is the limiting factor for G1 phase progression and S phase entryS phase entryRecently,several splice variants of cycli
17、n E1,which are not present in normal cells,have also been discovered;which stimulate cells to progress through the cell cycle much more efficiently than the full length cyclin E1Cyclin E was prognostic in seven out of 10 studies.Cyclin E was prognostic in seven out of 10 studies.u肿瘤细胞的生长肿瘤细胞的生长免疫组织化
18、学方法免疫组织化学方法细胞周期蛋白细胞周期蛋白The overexpression of cyclin E was accompanied by the The overexpression of cyclin E was accompanied by the appearance of low molecular weight(LMW)isoforms,and both appearance of low molecular weight(LMW)isoforms,and both were a reliable prognostic marker in stage IIII BC pati
19、ents.were a reliable prognostic marker in stage IIII BC patients.High levels of cyclin E1 were predictive of resistance to High levels of cyclin E1 were predictive of resistance to tamoxifen adjuvant therapy in 108 node-positive BC patients,tamoxifen adjuvant therapy in 108 node-positive BC patients
20、,independently of ER status.independently of ER status.Cyclin D1Cyclin D1u肿瘤细胞的生长肿瘤细胞的生长细胞周期蛋白细胞周期蛋白D-type cyclins are other key regulator proteins of the G1 D-type cyclins are other key regulator proteins of the G1 phase progression.phase progression.The protein is synthesized in response toThe pro
21、tein is synthesized in response togrowth factors;its levels reach a maximum in the mid-growth factors;its levels reach a maximum in the mid-G1phase of the cell cycle and then begin to drop.It appears G1phase of the cell cycle and then begin to drop.It appears that the association of cyclin D1 to Cdk
22、 is crucial to drive that the association of cyclin D1 to Cdk is crucial to drive cells to the restriction point where the cell is committed to cells to the restriction point where the cell is committed to dividedivideA strong correlation between overexpression of cyclin A strong correlation between
23、 overexpression of cyclin D1 and HR-positivity has been reported in the majority D1 and HR-positivity has been reported in the majority of trials,but cyclin D1 does not appear to be a strong of trials,but cyclin D1 does not appear to be a strong prognosticprognosticmarker.In fact,its overexpression
24、has been associated marker.In fact,its overexpression has been associated with better RFS in only one studywith better RFS in only one studyCyclin D1Cyclin D1u肿瘤细胞的生长肿瘤细胞的生长细胞周期蛋白细胞周期蛋白u肿瘤的生长与扩散肿瘤的生长与扩散l肿瘤的扩散方式肿瘤的扩散方式直接蔓延直接蔓延转移转移metastasismetastasis瘤细胞从原发部位瘤细胞从原发部位 侵入淋巴管、血管和体腔，侵入淋巴管、血管和体腔，扩散到其它部位，扩散
25、到其它部位，形成与原发瘤形成与原发瘤相同的肿瘤。相同的肿瘤。转移转移metastasismetastasisu肿瘤的生长与扩散肿瘤的生长与扩散肿瘤的扩散方式肿瘤的扩散方式淋巴道转移淋巴道转移Lymphatic metastasis is a predictor of poor outcome in Lymphatic metastasis is a predictor of poor outcome in many solid malignancies.many solid malignancies.The presence of lymph node metastases decreases
26、 the 5-The presence of lymph node metastases decreases the 5-year survival of melanoma patients independent of other year survival of melanoma patients independent of other prognostic factors of the primary tumor.prognostic factors of the primary tumor.Figure 1.Figure 1.Development of lymphatic vess
27、els in embryogenesis and cancerSome of the proteins that are important in theseevents are shown underneath each section.Arrows denote the direction of lymph flow in thelymphatic vessels.转移转移metastasismetastasisu肿瘤的生长与扩散肿瘤的生长与扩散肿瘤的扩散方式肿瘤的扩散方式血道转移血道转移转移转移metastasismetastasisu肿瘤的生长与扩散肿瘤的生长与扩散l肿瘤的扩散方式肿瘤
28、的扩散方式种植性转移种植性转移体腔内脏器的肿瘤蔓延至器官表面时，体腔内脏器的肿瘤蔓延至器官表面时，瘤细胞可脱落种植于瘤细胞可脱落种植于体腔和各器官表面形成多数转移瘤。体腔和各器官表面形成多数转移瘤。u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制肿瘤侵袭是转移的前提；肿瘤侵袭是转移的前提；侵袭和转移的步骤：侵袭和转移的步骤：脱离原发瘤群体脱离原发瘤群体向周围组织浸润向周围组织浸润与局部血管或淋巴管密切接触，与局部血管或淋巴管密切接触，穿过穿过其管壁其管壁穿透管壁，穿透管壁，在基质中增生在基质中增生转移灶的形成和生长转移灶的形成和生长u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏
29、附分子细胞黏附与细胞黏附分子侵袭和转移的步骤：侵袭和转移的步骤：脱离原发瘤群体脱离原发瘤群体以配体核受体结合的形式，以配体核受体结合的形式，使细胞间发生粘连使细胞间发生粘连integrinintegrin跨膜糖蛋白跨膜糖蛋白十六种十六种a a亚单位和亚单位和8 8种种b b亚单位亚单位u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏附分子细胞黏附与细胞黏附分子cadherincadherin与细胞骨架连接与细胞骨架连接人类至少有人类至少有1010多种钙粘蛋白多种钙粘蛋白E E；N N；P Pu肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏附分子细胞黏附与细胞黏附
30、分子IgSFIgSF跨膜蛋白跨膜蛋白具有与具有与IgIg类似的结构类似的结构u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏附分子细胞黏附与细胞黏附分子Selectin familySelectin familycancer cells adhere to by a process similar to that of LC cancer cells adhere to by a process similar to that of LC homing.In this model,cells in flow are captured on the homing.In this
31、model,cells in flow are captured on the endothelial surface.endothelial surface.transient adhesive interactions by cells with endothelial selectins(rolling),and firmly anchored on (firm adhesion)to enable entry into the underlying tissue.The selectins,particularly E-selectin,are recognized to mediat
32、e adhesion and thus potentiate of certain cancersu肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏附分子细胞黏附与细胞黏附分子Selectin familySelectin familyu肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制l细胞黏附与细胞黏附分子细胞黏附与细胞黏附分子CD44CD44A transmembrane protein Essential for the homing and properties of leukemicCells,CD44 has also been found to support anch
33、orage-independent growth in vitro and tumor growth and in experimental models of solid cancers u肿瘤细胞扩散的过程机制肿瘤细胞扩散的过程机制肿瘤细胞从原发灶分离的机制肿瘤细胞从原发灶分离的机制肿瘤细胞表面黏附分子减少。肿瘤细胞表面黏附分子减少。癌细胞钙含量降低癌细胞钙含量降低恶性肿瘤细胞间连接结构数量减少恶性肿瘤细胞间连接结构数量减少肿瘤细胞表面电荷增加肿瘤细胞表面电荷增加l肿瘤细胞向周围组织的浸润肿瘤细胞向周围组织的浸润细胞外基质的降解细胞外基质的降解瘤细胞的运动瘤细胞的运动趋化因子的作用趋化因子
34、的作用肿瘤血管生成肿瘤血管生成肿瘤血管生成肿瘤血管生成l肿瘤细胞向周围组织的浸润肿瘤细胞向周围组织的浸润肿瘤血管生成肿瘤血管生成肿瘤组织中微血管的来源肿瘤组织中微血管的来源瘤细胞生成的多种生长因子瘤细胞生成的多种生长因子诱导瘤体生成微血管诱导瘤体生成微血管残存于流体的宿主血管逐渐残存于流体的宿主血管逐渐变为肿瘤血管变为肿瘤血管VEGFVEGF是迄今鉴定出来的是迄今鉴定出来的最重要的血管生成因子最重要的血管生成因子Fig.1 Switching on the angiogenic phenotype in tumors by genetic and epigenetic factors.Both
35、 malignant and nonmalignant cells produce multiple angiogenic factors and cytokines to induce tumor neovascularization.Endogenous angiogenesis inhibitors are down regulated to support the angiogenic phenotype肿瘤细胞侵入血管和淋巴管肿瘤细胞侵入血管和淋巴管侵入血管和淋巴管侵入血管和淋巴管在循环中运行到达远处部位在循环中运行到达远处部位Fig.1.Schematic diagram show
36、ing how production of VEGF-C and VEGF-C in tumors can induce lymphangiogenesis,leading to increased lymphatic vessel density in the vicinity of the tumor,and subsequently to metastasis of invasive tumor cells via the lymph vessels.Fig.1.l转移灶的形成和生长转移灶的形成和生长u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因Nm23Nm23基因基因NM23-H1 a
37、nd NM23-H2 in humanNM23-H1 and NM23-H2 in humanNucleoside diphosphate kinases(NDPKs)catalyze the exchange of-phosphate between nucleoside(and 2-deoxynucleoside)triphosphates and diphosphates with formation of a high-energy phosphohistidine intermediate(Parks and Agarwal 1973).They are encoded by the
38、 NME genes(also known as NM23).参与调节细胞内微管系统的状态参与调节细胞内微管系统的状态高度表达高度表达nm23nm23表现为低转移属性表现为低转移属性u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因肿瘤转移相关基因肿瘤转移相关基因mtalmtalu肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因Tiam1 Tiam1 基因基因鼠鼠T T淋巴细胞瘤中克隆出来的基因。淋巴细胞瘤中克隆出来的基因。产物具有产物具有15911591个氨基酸残基，个氨基酸残基，把蛋白质锚定在质膜上把蛋白质锚定在质膜上TIAM1TIAM1 T-cell lymphoma invasion and
39、metastasis 1 Homo sapiens T-cell lymphoma invasion and metastasis 1 Homo sapiens Zhonghua Bing Li Xue Za Zhi.2009 Apr;38(4):268-72.Overexpression of Tiam1 gene and its relationship with invasive and metastatic ability of Overexpression of Tiam1 gene and its relationship with invasive and metastatic
40、ability of nasopharyngeal carcinoma.nasopharyngeal carcinoma.Article in ChineseArticle in ChineseZhang XM,Ding Y,Chen JZ,Jin H,Yu LN,Li YF,Ding YQ.Currently,many GEFs,including Vav1,LARG,Bcr and T-lymphoma invasion and metastasis 1(Tiam1),have been identified as oncogenes.RESULTS:RESULTS:Tiam1 over
41、expression significantly increased the abilities of Tiam1 over expression significantly increased the abilities of adhesion,migratory and invasion of C666-1 and CNE1 cells,adhesion,migratory and invasion of C666-1 and CNE1 cells,comparing with that of the control untransfected cells comparing with t
42、hat of the control untransfected cells(P 0.05).(P 0.05).CONCLUSION:CONCLUSION:Tiam1 expression correlates with the invasion and metastasis Tiam1 expression correlates with the invasion and metastasis of nasopharyngeal carcinoma cells.of nasopharyngeal carcinoma cells.u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因Tiam1 Tia
43、m1 基因基因鼠鼠T T淋巴细胞瘤中克隆出来的基因。淋巴细胞瘤中克隆出来的基因。Overexpression of Tiam1 in hepatocellular carcinomas predicts poor Overexpression of Tiam1 in hepatocellular carcinomas predicts poor prognosis of HCC patientsprognosis of HCC patientsYi Ding1,Bin Chen2,Shuang Wang3,Liang Zhao3,Juanzhi Chen3,Yi Ding1,Bin Chen2
44、,Shuang Wang3,Liang Zhao3,Juanzhi Chen3,Yanqing Ding3,Longhua Chen1Yanqing Ding3,Longhua Chen1*and Rongcheng Luo2 and Rongcheng Luo2*Int.J.Cancer:124,653658(2009)Int.J.Cancer:124,653658(2009)2008 Wiley-Liss,Inc.2008 Wiley-Liss,Inc.u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因Tiam1 Tiam1 基因基因鼠鼠T T淋巴细胞瘤中克隆出来的基因。淋巴细胞瘤中克隆出来的
45、基因。生长因子通过生长因子通过RasRas信号通路，信号通路，导致细胞增殖。导致细胞增殖。转染给转染给NIH3T3NIH3T3细胞，细胞，引起大量侵袭和转移。引起大量侵袭和转移。Activated KrasActivated KrasG12DG12D is associated with invasion and metastasis is associated with invasion and metastasis of pancreatic cancer cells through inhibition of E-cadherinof pancreatic cancer cells th
46、rough inhibition of E-cadherinBritish Journal of CancerBritish Journal of Cancer 104,1038-1048(15 March 2011)104,1038-1048(15 March 2011)S Rachagani,S Senapati,S Chakraborty,M P Ponnusamy,S Kumar,S Rachagani,S Senapati,S Chakraborty,M P Ponnusamy,S Kumar,L M Smith,M Jain and S K Batra L M Smith,M Ja
47、in and S K Batra u肿瘤侵袭和转移相关基因肿瘤侵袭和转移相关基因Ras Ras 基因基因包括包括H-rasH-ras，K-ras K-ras 和和N-ras N-ras 三类，三类，Results:The KrasG12D knockdown cells exhibited a significant decrease in motility(P0.0001),invasion(P0.0001),anchorage-dependent(P0.0001)and anchorage-independent growth(P0.0001),proliferation(P0.005)a
48、nd an increase in cell doubling time(P0.005)in vitro and a decrease in the incidence of metastases upon orthotopic implantation into nude mice.The knockdown of the KrasG12D allele led to a significant increase in the expression of E-cadherin(mRNA and protein)both in vitro and in vivo.This was associ
49、ated with a decrease in the expression of phoshpo-ERK-1/2,NF-B and MMP-9,/2,NF-B and MMP-9,and transcription factors such as EF1,Snail and ETV4.Furthermore,the and transcription factors such as EF1,Snail and ETV4.Furthermore,the expression of several proteins involved in cell survival,invasion and m
50、etastasis expression of several proteins involved in cell survival,invasion and metastasis was decreased in the Kraswas decreased in the KrasG12D knockdown cells.Conclusions:The results of this study suggest that the KrasG12D allele promotes metastasis in PC cells partly through the downregulation o
