肿瘤学 （ Oncology)

1. 肿瘤学 （Oncology)

2. 癌症(cancer) 肿瘤(tumor) Carcinoma 癌 恶性 良性 Sarcoma 肉瘤 Leukemia 白血病

3. 恶性肿瘤: 人类死亡的第一或第二元凶 现代医学面临的重要挑战之一 儿童肿瘤发病率在逐渐升高。在5-10岁儿童中，恶性肿瘤在造成死亡病种中排名第一位。 提高肿瘤防治水平和寻找治疗肿瘤的新方法已成为科学家和临床医学家所面临的最大挑战。 恶性肿瘤依然是现代医学面临的重要挑战之一。恶性肿瘤已经成为人类死亡的第一或第二元凶。

4. 多因素、多步骤

5. 第一节 肿瘤病因学

6. Chemical carcinogen Pott P(1775): 童年时当过烟囱清扫工的男性患阴囊癌的比率增高 Volkman and Bell (1870’s): 长期与石蜡油和焦油接触的工人易患皮肤癌 Rehn (1880’s): 接触苯胺的工人发生泌尿道膀胱肿瘤 Yamagiwa and Ichikawa (1915): 反复用煤焦油涂擦兔耳成功地诱发了皮肤癌 Cook James (1933): 证明多种化学致癌物(Benzopyrene) 与动物肿瘤的关系

7. Pott P(1775): Volkman and Bell (1870’s): Chemical carcinogen Rehn (1880’s): Yamagiwa and Ichikawa (1915): Cook James (1933): 进入机体后，无需代谢活化即有致癌作用的化学致癌物 单独作用无致癌作用，但对其它致癌物具有促进作用 β-丙内酯、硫酸二甲酯、氮芥、溶肉瘤素、亚硝酸胺类、二（氯甲）醚等 巴豆油、糖精、 苯巴比妥 Pro- carcinogen direct indirect 进人体内后，必需经过体内代谢活化，才具致癌作用 多环芳香烃、芳香胺类、亚硝胺、黄曲霉素

8. Indirect Carcinogen 前致癌物 未经代谢活化的、不活泼的间接致癌物 precarcinigen 近致癌物 经代谢转变为化学性质活泼、 寿命极短的致癌物 Proximate carcinigen 终致癌物 带正电荷的亲电子物质 Ultimate carcinigen

9. Indirect Carcinogen precarcinigen Proximate carcinigen Phase II enzymes: act on oxidized substrates methyltransferases, acetyltransferases, glutathione transferases, uridine 5'-diphosphoglucuronosyl transferases, Ultimate carcinigen sulfotransferases, nicotinamide-adenine dinucleotide (NAD)- and nicotinamide-adenine dinucleotide phosphate (NADP)-dependent alcohol, aldehyde and steroid dehydrogenases, quinone reductases, NADPH diaphorase, azo reductases, aldoketoreductases, transaminases, esterases, and hydrolases. • Cytochrome P-450 (CYP) : phase I enzymes • act by adding O atom onto the substrate • inducible by polycyclic aromatic and chlorinated hydrocarbons. • largely responsible for the metabolic activation and detoxication.

10. Phase II enzymes: act on oxidized substrates methyltransferases, acetyltransferases, glutathione transferases, uridine 5'-diphosphoglucuronosyl transferases, sulfotransferases, nicotinamide-adenine dinucleotide (NAD)- and nicotinamide-adenine dinucleotide phosphate (NADP)-dependent alcohol, aldehyde and steroid dehydrogenases, quinone reductases, NADPH diaphorase, azo reductases, aldoketoreductases, transaminases, esterases, and hydrolases. • Cytochrome P-450 (CYP) : phase I enzymes • act by adding O atom onto the substrate • inducible by polycyclic aromatic and chlorinated hydrocarbons. • largely responsible for the metabolic activation and detoxication.

11. Indirect Carcinogen precarcinigen Proximate carcinigen Ultimate carcinigen carcinogen DNA adducts

12. Metabolic activation detoxication Polycyclic arimatic hydrocarbon, PAH Tobacco DNA damage Tobacco-specific nitrosamines,TSNA DNA repair Aromatic amines malignant 代谢激活速率高而代谢解毒率低 或DNA修复功能低下的个体肿瘤易感性高

13. virus carcinogen Ellermann and Bang(1905,丹麦 ): chicken leukemia DNA virus 用无细胞的滤液首次证实病毒与恶性肿瘤有病因学上的关系 Rous (1911，美国): Rous sarcoma virus (RSV) 用滤液成功地诱发了鸡的肉瘤 Shope (1933): 将病毒所致的野兔乳头状瘤进行皮下移植实验， 发生浸润性鳞癌 Burkitt (1962): viruslymphoma RNA virus 发现病毒可以引起淋巴瘤 Epstein and barr (1964): EB virus 在Burkitt淋巴瘤细胞培养液中发现EB病毒

14. DNA virus 与动物或人类肿瘤有关的致瘤性NDA病毒： • 乳-多-空病毒类 SV40 T抗原 HPV16、HPV18 E6、E7转化蛋白 • 疱疹病毒类 • 腺病毒类 转化蛋白E1A、E1B • 乙型肝炎病毒类 核蛋白 p53 • 痘病毒类 Rb 共同特征： • 病毒的核心是由DNA和蛋白质组成的复合体 • DNA病毒一般没有细胞内同源物 • 病毒的致癌作用发生在病毒进入细胞后复制的早期阶段，相关的瘤基因多整合至宿主细胞DNA上。 某些NDA病毒在染色体上的定位具有倾向性，往往表现为累及多个染色体的位点，可能涉及到染色体的脆性部位和原瘤基因的位点.

15. 早期:产生转化蛋白 晚期:形成病毒颗粒 细胞裂解 DNA virus DNA virus DNA virus permissive cell DNA virus DNA virus DNA virus DNA virus lytic infection DNA virus DNA virus DNA virus abortive infection DNA virus DNA virus 基因组整合到细胞的DNA中， 使细胞发生转化 Non-permissive cell

16. RNA RNA LTR LTR env gag pol RNA virus retrovirus

17. provirus

18. 1 3 2 LTR LTR 2 1 3 env gag pol C-src 60KD、具有酪氨酸激酶活性的PP60蛋白，参与信号转导途径，与多种肿瘤发病相关 LTR LTR env gag pol Non-defective virus the Rous sarcoma virus (RSV): the RSV transforming gene (designated v-src) was homologous to a host cellular gene (c-src)

19. 1 3 2 LTR LTR 2 1 3 pol gag LTR LTR env gag pol defective virus

20. Acutely transforming retroviruses can rapidly cause tumors within days after injection. These retroviruses can also transform cell cultures to the neoplastic phenotype. Chronic or weakly oncogenic retroviruses can cause tissue-specific tumors in susceptible strains of experimental animals after a latency period of many months. Although weakly oncogenic retroviruses can replicate in vitro, these viruses do not transform cells in culture.

21. 染色体水平 种族分布差异 遗传因素 家族聚集现象 遗传缺陷 流行病学分析 家系分析 细胞遗传学研究 分子遗传学研究 分子/基因水平

22. Knudson： Chromosomal mechanisms which result in loss of heterozygosity for alleles at the retinoblastoma predisposition (RB1) locus Retinoblastoma the most common intraocular cancer in children Incidence ： 1 of 13,500 - 25,000 live births Sites: unilateral (20% hereditary), bilateral (all hereditary) Onset time: Bilateral -12 m,unilateral- 18 m, Most (90%) : < 3 y

23. 遗传性肿瘤综合征 具有恶变倾向的癌前病变（多发性、良性） Li-Fraumeni综合征： 易发软组织肿瘤、乳腺癌、脑瘤 遗传基础：P53基因生殖细胞突变 遗传易感基因

24. DNA recombinant techniques DNA transfection Karyotypic analysis Molecular clone Oncogene

25. 慢性粒细胞性白血病（CML）

27. Bcr-abl 酪氨酸激酶 激酶底物

28. DNA recombinant techniques DNA transfection Karyotypic analysis Molecular clone Tumor suppressor mutator / DNA-mismatch repair -involved genes hereditary nonpolyposis colorectal cancer (HNPCC) syndromes homogeneously staining regions [HSR] Oncogene Oncogene Oncogene Tumor suppressor mutator / DNA-mismatch repair -involved genes

29. 第二节：癌基因 （Oncogene） Oncogenes are essential for human life activity, whose normal function is to control cellular growth and differentiation /apoptosis or, in different terms, cell birth and cell death. Correspondingly, their structural and/or functional alterations lead uncontrolled cellular growth and abnormal differentiation/apoptosis

30. 3T3 cells

31. 生长因子 生长因子受体 信号传递分子 转录因子

32. insersion 30%:lung adenocarcinomas 50% colon carcinomas 90%:pancreatic carcinomas deletion K-ras carcinomas 15-20% ORF RS Ras N-ras Point mutation H-ras Blood malignancies Exon 12 ORF RS RS Mechanisms of Oncogene Activation ORF RS ORF RS mutation amplification Proto-oncogene ORF RS Re-arrangement

33. t(4;11)(q21;q23) in ALL q32 q32 t(8;14)(q24;q32) in B-cell ALL and Burkitt's lymphoma 14 14 t(1;19)(q21;p13) in pre-B cell ALL q32 q32 t(14;18)(q32;q21) in B-cell NHL 14 14 inv(14)9q11q32) in T-cell leukemia/lymphoma 14 14 t(8;14)(q24;q11) in T-cell leukemia/lymphoma q32 q32 8q24 :MYC gene 14q32 : IGH 18q21: BCL2 Partial karyotypes of trypsin-Giemsa-banded metaphase cells depicting nonrandom chromosomal rearrangements observed in lymphoid malignant diseases.

34. ras Family. The ras family of oncogenes (homologous to the rat sarcoma virus) has three primary members (H-ras, K-ras, and N-ras) which are among the most common activated oncogenes found in human cancer. The ras genes code for a protein (p21) that is located on the inner surface of the plasma membrane, has GTPase activity, and may participate in signal transduction. ras oncogenes are activated by point nucleotide mutations that alter the amino acid sequence of p21.

35. ras in Carcinogen-Induced Tumors • Studies in mice with carcinogen-induced lung cancers implicate genes of the ras family in the carcinogenesis process. Mouse lung tumors induced by tetranitromethane contained mutated K-ras genes. • Mice harboring the mutated H-ras transgene developed tumors exclusively in the lungs within weeks following birth. • Ninety percent of these tumors had transforming genes in the (NIH) 3T3 assay; the gene was K-ras in all lung tumors.

36. Human Studies. • In most studies, K-ras mutations were confined to adenocarcinomas of the lung and occurred in 30% of tumors. Mutations were not observed in adenocarcinomas from nonsmokers. • Studies done so far favor the interpretation that ras activation contributes to progression in lung cancer. ras apparently is activated in about one-third of adenocarcinomas arising in patients with a history of heavy smoking. However, premalignant lung lesions have not been studied to determine if such mutations exist at the precancerous stage, as is the case for adenocarcinoma of the colon. • ras mutations can influence the differentiation of tumor cells. For example, infection of SCLC cell lines with the Harvey murine sarcoma virus alters the phenotype of cells.Following infection, the variant SCLC cell line developed features of a large cell undifferentiated lung carcinoma, including increased carcinoembryonic antigen and keratin expression. • K-ras mutation appears to be an independent prognostic factor that indicates a poor prognosis and is unrelated to conventional staging criteria, such as tumor size or lymph node metastases. In all patients studied, mutations occurred in a single allele. is the case for adenocarcinoma of the colon.

37. Antisense/iRNA/ribozyme technology: Antisense was used to study the effects of eliminating expression of a mutant K-ras oncogene in NSCLC cells. A selective decrease in mutant oncogene expressions was achieved, associated with decreased growth of cancer cells as heterografts in nude mice. This technique provides an opportunity to determine the effects of selective inhibition of oncogenic protein expression on the malignant phenotype. This study and subsequent confirmatory ones showed that reversal of a single genetic abnormality was sufficient to prevent human cancer cells from forming tumors. Thus, it may not be necessary to reverse all the genetic lesions in a cancer cell to achieve a therapeutic effect.

38. Antisense/iRNA/ribozyme technology: Antisense was used to study the effects of eliminating expression of a mutant K-ras oncogene in NSCLC cells. A selective decrease in mutant oncogene expressions was achieved, associated with decreased growth of cancer cells as heterografts in nude mice. This technique provides an opportunity to determine the effects of selective inhibition of oncogenic protein expression on the malignant phenotype. This study and subsequent confirmatory ones showed that reversal of a single genetic abnormality was sufficient to prevent human cancer cells from forming tumors. Thus, it may not be necessary to reverse all the genetic lesions in a cancer cell to achieve a therapeutic effect.

39. These observations raise the intriguing possibility of specific gene therapy for cancer. These observations raise the intriguing possibility of specific gene therapy for cancer. Gene sequences could be delivered to tumor cells via viral vectors that specifically inhibit expression of the oncogenes activated in the cancer cell. Such constructs would be relatively nontoxic because, as in the example above, they could target a single gene whose function may be assumed by other redundant genes of the same family.

40. 第三节：抑癌基因 （tumor suppressor gene） （antioncogene ） 这类基因作为细胞的刹车而起作用。它们编码的蛋白能够抑制细胞的生长，阻止细胞恶性转变。 如果其功能失活或出现基因缺失、突变等异常，将导致细胞恶性转化而形成肿瘤。

41. 抑癌基因的生物学功能与癌基因相反，是机体细胞在身生长、增殖、分化和凋亡等生命过程中的负调控信号。确定一种抑癌基因在理论上需符合三个基本条件：抑癌基因的生物学功能与癌基因相反，是机体细胞在身生长、增殖、分化和凋亡等生命过程中的负调控信号。确定一种抑癌基因在理论上需符合三个基本条件： ①该基因在与恶性肿瘤组织相对应的正常组织中必需正常表达； ②恶性肿瘤中这种基因应有功能失活、结构改变或表达缺陷； ③将这种基因的野生型导入基因异常的肿瘤细胞内，可部分或全部逆转肿瘤的恶性表型。

42. Knudson：two hit hypothesis 遗传性肿瘤发病早，肿瘤表现为多发性或双侧性 散发性肿瘤发病迟，肿瘤表现为单发性或单侧性

43. 1983年Cavenee等 loss of heterozygosity，LOH 染色体 13q14处的DNA标志。 1978年 Francke 13q14 • 1986年，美国三个实验室分别独立克隆了该基因 Rb

44. G2 M G1 S Quiescence (G0) Cell cycle

45. CDK CDKI cyclins G2 Cell cycle M G1 S CDK cyclins cyclins

46. Rb P Cyclin D1 CDK4 G2 Cell cycle M Rb Rb G1 P P P P P S P E2F DP DNA synthesis-related genes DNA synthesis-related genes E2F P P P E2F P E2F E2F E2F E2F E2F DP

47. Rb基因的异常主要表现为等位基因缺失和基因突变。除了视网膜母细胞瘤和骨肉瘤外，在许多种肿瘤中也检测到Rb基因的突变，包括肺癌（40%）, 膀胱癌（30%），前列腺癌（20%）以及乳腺癌，宫颈癌和某些类型的白血病。这些肿瘤中Rb基因的突变主要集中于外显子13~17上，并且主要表现为移码突变、终止码突变和点突变。其中一些突变能够影响第393-572位氨基酸以及第646-772位氨基酸之间的结构域。

49. 陈国强， 博士， 研究员。 1996年毕业于上海第二医科大学，获医学博士学位。 1997年和1999-2001年分别在法国和美国从事合作研究。现任上海第二医科大学附属瑞金医院Terry Fox肿瘤中心主任，上海血液学研究所副所长，上海第二医科大学病理生理学教研室主任，中国科学院上海生命科学研究院健康科学中心肿瘤功能基因组学研究室主任，国家重点基础研究计划首席科学家，上海第二医科大学肿瘤合作研究学术委员会主任、《癌症》杂志副主编、《中华医学杂志》《中国肿瘤生物治疗杂志》《肿瘤防治杂志》等编委、《BLOOD》和《LEUKEMIA》杂志审稿人。 陈国强， 博士， 研究员。 1996年毕业于上海第二医科大学，获医学博士学位。 1997年和1999-2001年分别在法国和美国从事合作研究。现任上海第二医科大学附属瑞金医院Terry Fox肿瘤中心主任，上海血液学研究所副所长，上海第二医科大学病理生理学教研室主任，中国科学院上海生命科学研究院健康科学中心肿瘤功能基因组学研究室主任，国家重点基础研究计划首席科学家，上海第二医科大学肿瘤合作研究学术委员会主任、《癌症》杂志副主编、《中华医学杂志》《中国肿瘤生物治疗杂志》《肿瘤防治杂志》等编委、《BLOOD》和《LEUKEMIA》杂志审稿人。 功能基因组学、肿瘤细胞蛋白质组学及药物靶标的发和功能研究方面取得多项原创性发现。他在国际权威刊物《BLOOD》、《JNCI》、《PNAS》、《LEUKEMIA》以及国内核心期刊共发表了70余篇 功能基因组学、肿瘤细胞蛋白质组学及药物靶标的发和功能研究方面取得多项原创性发现。他在国际权威刊物《BLOOD》、《JNCI》、《PNAS》、《LEUKEMIA》以及国内核心期刊共发表了70余篇 自1993年以来，他一直致力于肿瘤尤其是白血病细胞凋亡、分化机制和治疗学基础研究，尤其是他所从事的三氧化二砷治疗急性早幼粒细胞性白血病（APL）的基础和临床研究工作在国际血液学界开拓了一种全新的研究领域。在肿瘤 自1993年以来，他一直致力于肿瘤尤其是白血病细胞凋亡、分化机制和治疗学基础研究，尤其是他所从事的三氧化二砷治疗急性早幼粒细胞性白血病（APL）的基础和临床研究工作在国际血液学界开拓了一种全新的研究领域。在肿瘤 学术论文。自97年以来，在全国单篇论文引用数统计排名中始终处于前八位，共达1400余次。他先后获得国家杰出青年科学基金、中科院百人计划、国家973计划和上海科技发展重大项目的资助，并获得国家自然科学二等奖、上海市科技进步一等奖、杜邦科技创新奖、中国青年科技奖等学术奖励和新世纪百千万人才工程首批国家级人选、上海“十大杰出青年”、上海青年科技英才、上海市劳动模范、上海优秀青年教师、市卫生系统杰出青年人才 “银蛇奖”、上海市优秀学科带头人等荣誉称号，并享受国务院特殊津贴 。 学术论文。自97年以来，在全国单篇论文引用数统计排名中始终处于前八位，共达1400余次。他先后获得国家杰出青年科学基金、中科院百人计划、国家973计划和上海科技发展重大项目的资助，并获得国家自然科学二等奖、上海市科技进步一等奖、杜邦科技创新奖、中国青年科技奖等学术奖励和新世纪百千万人才工程首批国家级人选、上海“十大杰出青年”、上海青年科技英才、上海市劳动模范、上海优秀青年教师、市卫生系统杰出青年人才 “银蛇奖”、上海市优秀学科带头人等荣誉称号，并享受国务院特殊津贴 。 Chengq@shsmu.edu.cn Thanks