癌基因与抑癌基因
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癌生物学课程
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
百度文库
➢ a genetic disease characterized by uncontrolled cell growth
gag, pol, env
unhybridized sequences
hybridized sequences
src
gag, pol, env
Cont.
Src （v-onc, viral oncogene） probe
RSV-Infected CEF (+ control)
“Normal” chick DNA
D Activated Proto-oncogenes from DNA transfection
Human Bladder Tumor cell line DNA
Isolate high MW DNA
Restriction endonuclease
Isolate DNA fragments
Transfection
RESULT: The gene is the HUMAN c-H-ras gene
Compare sequence to NORMAL gene
RESULT: RAS “Activation” is due to a SINGLE point mutation (gly val) at codon 12
Mouse
Drosophila
Human
+
+
+
+
+
Thus: a proto-oncogene is the NORMAL progenitor gene of a viral oncogene
1980, 1st oncogene identified
1989 Nobel Prize for Bishop and Varmus
1st human oncogene mutation Ras G12V was identified !
1982, Robert Weinberg, etc three groups
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 in the appropriate animal. V-onc is encoded by cellular sequences that have become inserted into the viral genome.
B
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
Proto-oncogene (c-onc): a normal cellular form of a gene that controls cell
proliferation and can be converted into a cancer-promoting gene by mutation, whose continued activation leads to continued signal transduction, and whose aberrant expression or activity may contribute to tumorigenesis .
NIH 3T3 fibroblasts
Transformation
Isolate DNA (>99% mouse + 8-10 human genes)
Transformation
Identify Human DNA
Cont.
Use Alu probe Isolate Human DNA
RESULT: A SINGLE human gene is responsible for transforming capability Sequencing
一、癌基因 Oncogene
1.癌基因的发现
Virus and Cancer
A Non-Transforming (Avian Leukemia Viruses, ALV)
Ellerman and Bang, 1908 1, Induce leukemia after long latency periods 2, Do not “transform” tissue culture cells
C Isolation of Retroviral Transforming Genes
RSV (gag, pol, env, src)
RSV (td: gag, pol, env)
R.T.(1975 Nobel)
cDNA
genomic RNA
gag, pol env src
Denature and Hybridize 1975,1977
Oncogene: an altered gene whose product can act in a dominant fashion to help
make a cell cancerous. Typically, an oncogene is a mutant form of a normal gene (proto-oncogene) involved in the control of cell growth or division.
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
百度文库
➢ a genetic disease characterized by uncontrolled cell growth
gag, pol, env
unhybridized sequences
hybridized sequences
src
gag, pol, env
Cont.
Src （v-onc, viral oncogene） probe
RSV-Infected CEF (+ control)
“Normal” chick DNA
D Activated Proto-oncogenes from DNA transfection
Human Bladder Tumor cell line DNA
Isolate high MW DNA
Restriction endonuclease
Isolate DNA fragments
Transfection
RESULT: The gene is the HUMAN c-H-ras gene
Compare sequence to NORMAL gene
RESULT: RAS “Activation” is due to a SINGLE point mutation (gly val) at codon 12
Mouse
Drosophila
Human
+
+
+
+
+
Thus: a proto-oncogene is the NORMAL progenitor gene of a viral oncogene
1980, 1st oncogene identified
1989 Nobel Prize for Bishop and Varmus
1st human oncogene mutation Ras G12V was identified !
1982, Robert Weinberg, etc three groups
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 in the appropriate animal. V-onc is encoded by cellular sequences that have become inserted into the viral genome.
B
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
Proto-oncogene (c-onc): a normal cellular form of a gene that controls cell
proliferation and can be converted into a cancer-promoting gene by mutation, whose continued activation leads to continued signal transduction, and whose aberrant expression or activity may contribute to tumorigenesis .
NIH 3T3 fibroblasts
Transformation
Isolate DNA (>99% mouse + 8-10 human genes)
Transformation
Identify Human DNA
Cont.
Use Alu probe Isolate Human DNA
RESULT: A SINGLE human gene is responsible for transforming capability Sequencing
一、癌基因 Oncogene
1.癌基因的发现
Virus and Cancer
A Non-Transforming (Avian Leukemia Viruses, ALV)
Ellerman and Bang, 1908 1, Induce leukemia after long latency periods 2, Do not “transform” tissue culture cells
C Isolation of Retroviral Transforming Genes
RSV (gag, pol, env, src)
RSV (td: gag, pol, env)
R.T.(1975 Nobel)
cDNA
genomic RNA
gag, pol env src
Denature and Hybridize 1975,1977
Oncogene: an altered gene whose product can act in a dominant fashion to help
make a cell cancerous. Typically, an oncogene is a mutant form of a normal gene (proto-oncogene) involved in the control of cell growth or division.