癌基因与抑癌基因.pptx

癌基因与抑癌基因Oncogene Tumor Suppressor Gene 提 纲 一、癌基因 二、抑癌基因 三、癌基因/抑癌基因与多步骤癌变 四、癌基因/抑癌基因的鉴定 肿瘤发生机理 物理因子 化学因子 生物因子，如病毒 遗传物质改变 正常细胞 癌细胞 a genetic disease characterized by uncontrolled cell growth 一、癌基因 Oncogene 1.癌基因的发现 Non-Transforming (Avian Leukemia Viruses, ALV) Ellerman and Bang, 1908 1, Induce leukemia after long latency periods 2, Do not “transform” tissue culture cells Virus and Cancer Transforming (Sarcoma Viruses, RSV) Peyton Rous, 1911 -cell free lysates could induce sarcomas in other chickens 1，Acute; 2，Transform cultured cells 病毒致癌-- Nobel Prize! 1966 Isolation of Retroviral Transforming Genes RSV (gag, pol, env, src) R.T.(1975 Nobel) gag, pol env src cDNA RSV (td: gag, pol, env) gag, pol, env Denature and Hybridize 1975,1977 unhybridized sequences hybridized sequences gag, pol, env src genomic RNA probe RSV-Infected CEF (+ control) “Normal” chick DNA Mouse Drosophila Human + + + + + 1989 Nobel Prize for Bishop and Varmus Thus: a proto-oncogene is the NORMAL progenitor gene of a viral oncogene Cont. 1980, 1st oncogene identified Src （v-onc, viral oncogene） Human Bladder Tumor cell line DNA Isolate high MW DNA Isolate DNA fragments Restriction endonuclease Transfection NIH 3T3 fibroblasts Transformation Isolate DNA (99% mouse + 8-10 human genes) Transformation Identify Human DNA Activated Proto-oncogenes from DNA transfection RESULT: RAS “Activation” is due to a SINGLE point mutation (gly val) at codon 12 Use Alu probe Isolate Human DNA RESULT: A SINGLE human gene is responsible for transforming capability Sequencing RESULT: The gene is the HUMAN c-H-ras gene Compare sequence to NORMAL gene 1st human oncogene mutation Ras G12V was identified ! 1982, Robert Weinberg, etc three groups Cont. 2.癌基因定义 Definition Viral Oncogene (v-onc): a gene carried by a tumor virus (RNA or DNA), the expression of which is necessary and sufficient to induce transformation in tissue culture cells and tumors