Liyan Xu

1. Oncogene Cancer suppressive gene Liyan Xu

2. The basic concept 1 oncogene: • These genes code for proteins that are capable of stimulating cell growth and division. • In normal tissues and organisms, such growth-stimulating proteins are regulated, so that growth is appropriately limited. • However, changes/mutation in these genes may result in loss of growth regulation, leading to uncontrolled cell proliferation and tumor development. • These changed genes are known as oncogenes, because they induce the oncogenic state — cancer. • Oncogenes are dominant, because a change/mutation of only one of the cell’s two copies of that gene can lead to tumor formation.

3. The basic concept 2 proto-oncogenes: • The normal precurcers of these above genes are termed proto-oncogenes and are essential for normal cell growth and differentiation. • proto-oncogenes is also called as cellular oncogenes. 3 virus oncogene: • these genes are in viruses, may leading to uncontrolled cell proliferation and tumor development. • Virus oncogenes are homolog with that corresponding cellular oncogenes.

4. The basic concept 4 cancer suppressive genes: • These genes code for proteins whose normal function is to turn off cell growth. • A change/mutation in one of these growth-limiting genes may result in a protein product that has lost its growth limiting ability. • The normal forms of such gense have been shown to suppress tumor growth and are known as tumor suppressor genes or anti-oncogenes as well. • Because both cellular copies of a tumor suppressor gene must be mutated to foil its growth-limiting action, these genes are recessive in nature.

5. Fig. 20-1 Structure of RSV genome 9392 bp wide type viral genes oncogene pol env gag src LTR LTR to initiate and regulate transcription tyrosine kinase 526 residue 60 kD

6. The pp60V-SRC is anchored to the plasma membrane via an N-terminal myristyl group

7. Fig. 20-2 Integration of RNA virus genome and host cell genome RNA virus host cell virus RNA provirus DNA + c-onc. v-onc. + +

8. Mechanism on oncogene activated • to obtain a strong promoter or enhancer. • gene translocation. • protooncogene amplification in the genome. • point mutation • to obtain a strong promoter or enhancer.

9. avian leukemia virus genome ssRNA host cell genome c-myc LTR dsDNA to increase c-myc gene expression, 30-100 times, compare with no infection.

10. Mechanism on oncogene activated • to obtain stronge a promoter or a enhancer. • gene translocation. • protooncogene amplification in the genome. • point mutation

11. Burkit lymphoma 8 chromosome c-myc 14 chromosome IgH gene regulation region t（8:14） to increase c-myc gene expression obviously

12. Mechanism on oncogene activated • to obtain stronge a promoter or a enhancer. • gene translocation. • protooncogene amplification in the genome. • point mutation

13. ras or c-myc amplification 1 2 3 n expression of ras or c-myc is increased obviously

14. Mechanism on oncogene activated • to obtain stronge a promoter or a enhancer. • gene translocation. • protooncogene amplification in the genome. • point mutation

15. H-ras DNA Protein Gly GGC normal Val GTC carcinoma

16. 418 Fig. 20-3 Oncogene and signal transduction for growth EGF SIS extracellular erbB-2 PDGF receptor membrane PIP2 cytoplasm DG PLC RAS/Gpro. AC PKC SRC + AA cAMP PLA2 kinase ER PG kinase kinase PKA IP3 Ca2+ effects targets targets targets targets effects nucleus DNA mRNA RNA pol.

19. 418 Fig. 20-3 Oncogene and transduction of information for growth EGF SIS extracellular erbB-2 PDGF receptor membrane PIP2 cytoplasm DG PLC RAS/Gpro. AC PKC SRC + AA cAMP PLA2 kinase ER PG kinase kinase PKA IP3 Ca2+ effects targets targets targets targets effects nucleus DNA mRNA RNA pol.

20. Phospho- rylation site

21. 418 Fig. 20-3 Oncogene and transduction of information for growth EGF SIS extracellular erbB-2 PDGF receptor membrane PIP2 cytoplasm DG PLC RAS/Gpro. AC PKC SRC + Ca2+ AA cAMP PLA2 kinase ER PG kinase kinase PKA IP3 Ca2+ effects targets targets targets targets effects nucleus DNA mRNA RNA pol.

23. 418 Fig. 20-3 Oncogene and transduction of information for growth EGF SIS extracellular erbB-2 PDGF receptor membrane PIP2 cytoplasm DG PLC RAS/Gpro. AC PKC SRC + AA cAMP PLA2 kinase ER PG kinase kinase PKA IP3 Ca2+ effects targets targets targets targets effects nucleus DNA mRNA RNA pol.

25. tumor suppressor genes • two examples • Rb: 13q14.1-14.2, 27 exon, complete length 180 kb, mRNA 4.7 kb, encoding protein 105 kD, 928 residues, locate in nucleus, inactive form: phosphorylation Rb, active form: non-phosphorylation Rb, be able to bind activated transcription factor E-2F, be able to inhibite RNA polymerase I and RNA polymerase III . • p53: 17p13, 11 exon, complete length 16-20 kb, mRNA 2.8 kb, encoding protein 53 kD, 393 residues, locate in nucleus, is phosphorylated, tetramer.

26. cell DNA damage p53 damaged DNA repair repair is unsuccessful by p53 repair is successful by p53 cell survive cell apoptosis

27. Oncogene and tumor suppresor gene  Our experiment result about the oncogenes and tumor suppressor genes for esophagealcancer 12-o-tetradecanoylphorbol -13-acetate,TPA carcinogen human papilloma virus E6 and E7 genes normal embryo esophageal epithelial cell immortal esophageal cell esophageal cancer cell plasmid transfection canceration cDNA array up clone: 33 down clone: 28 mRNA difference down up suppression subtractive hybridization up clone :62 down clone:56 difference cDNA clone

28. Summary • The basic concept: oncogene,proto-oncogene, virus oncogene, tumor suppressor gene • Mechanism on oncogene activated • Function of oncogene and tumor suppressor gene

29. 选择题练习 癌基因与抑癌基因

30. 1. 细胞原癌基因 A 正常人细胞也检测到的癌基因 B 只在肿瘤细胞中出现 C 加入化学致癌物到正常细胞后才出现 D 是细胞经过转化才出现 E 感染致癌病毒才出现

31. 2. 关于细胞癌基因,正确的叙述是 A 存在于RNA病毒中 B 存在于DNA病毒中 C 存在于正常细胞基因组中 D 正常细胞出现可导致肿瘤 E 只存在于肿瘤细胞细胞

32. 3. 致癌病毒 A 使人体直接致癌 B 使正常细胞转化为癌细胞 C 均为DNA病毒 D 均为RNA病毒 E 含转化酶

33. 4. 不属于癌基因产物的是 A 化学致癌物 B 生长因子类似物 C 结合GTP的蛋白质 D 结合DNA的蛋白质 E 酪氨酸蛋白激酶

34. 5. 有关抑癌基因,错误的叙述是 A 能抑制细胞的分化 B 能抑制细胞过度生长 C 突变后可导致肿瘤形成 D 可诱导细胞凋亡 E 最早发现的抑癌基因是Rb

35. 6. 有关P53蛋白,错误的描述是 A 其基因位于17p13, 突变后可致癌 B 能引发修复失败的细胞程序自杀 C 有“基因卫士”的称号 D 有转录因子作用 E 能激活解链酶

36. 7. 有关Rb基因,错误的描述是 A 位于13q14 B 是一种抑癌基因 C 其作用与E-2F有关 D 其编码蛋白为P21 E 突变后可导致肿瘤发生

37. 8. 关于凋亡,正确的叙述是 A 是一种病理过程 B 是细胞坏死 C 由意外事件引起的细胞损伤造成 D 由基因控制的细胞自我消亡的过程 E 与癌基因的表达调控无关

38. 9. 生殖细胞抑癌基因的突变可引起 A 生殖功能障碍 B 各种先天性的肿瘤 C 家族性的肿瘤易感性 D 后代的发育正常 E 家族性癌症发病率下降

39. 10. 使癌基因活化的因素有 A 正常基因表达增强 B 正常基因表达减少 C 抑癌基因表达增强 D 细胞增殖、分化加强 E 点突变

40. 11. The oncogene which can encode epidermal growth factor receptor is A src B ras C myc D sis E erb-B

41. 12. The oncogene which can encode GTP binding protein is A src B ras C myc D sis E erb-B

42. 13. The oncogene which can encode protein having TPK activity is A src B ras C myc D sis E erb-B

43. 14. Which isn’t a anti-oncogene? A P16 B P53 C Rb D WT1 E H-ras

44. 15. 正常基因的异常表达可致 A 细胞形态改变 B 细胞癌变 C 异常表型 D 细胞结构与生物活性改变 E 细胞凋亡

45. 16. 细胞癌基因可在下列情况下激活 A 基因发生突变 B 有化学致癌剂存在 C 有病毒感染 D 生长因子与DNA互相结合 E 受体激活

46. 17. 野生型P53基因 • 是抑制基因 • 一定情况下,能启动细胞自杀 • 编码蛋白有转录因子作用 • 能抑制解链酶活性 • 与复制因子A相互作用,参与DNA的复制与修复

47. 18. Rb基因的特点 A 位于13q14 B 含27个外显子 C 编码产物 105KD D 转录产物 4.7kb E 全长 20 kb

48. 19. Which is oncogene expression product? A GTP binding proteins B DNA binding proteins C tyrosine protein kinase D growth factors E cytoskeletal proteins

49. 20. The activation ways of proto-oncogene are A point mutation B gene translocation C gene amplification D protein phosphorylation E obtaining promoter