Oncogene and

Tumor Supressor Gene

Harliansyah, Ph.D
YARSI University
Department of Biochemistry

harliansyah.hanif @ yarsi.ac.id

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Cancer Pathogenesis

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 Cellular Basis of Cancer
Cancer is a collection of diseases
characterized by abnormal and
uncontrolled growth

Cancer arises from a loss of normal

growth control

In normal tissues, the rates of new

cell growth and old cell death are
kept in balance

In cancer, this balance is disrupted

This disruption can result from

1) uncontrolled cell growth or
2) loss of a cell's ability to undergo
apoptosis
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Cancer Cell Do Not Grow Faster Than
Normal Cells
Rather, Their Growth is Just
Uncontrolled

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1fertilizedegg
50x1012

1016celldivisions/lifetime

Proliferation Differentiation Death

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 Cellularequilibrium
Proliferation Differentiation Death

Transit
Renewing
Proliferating

Exiting 9
 Cancer:disruptionof
cellularequilibrium

Proliferation Differentiation Death

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Stemcellsasthetargetofcarcinogens
Postmitotic
Stemcell
Differentiated Normal
senescent
differentiated
cell
Benign
tumor
Grade2
malignancy
Grade3or4
malignancy
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Invasion and Metastasis
Abnormal cells proliferate
and spread (metastasize) to
other parts of the body

Invasion - direct
migration and
penetration into
neighboring tissues

Metastasis - cancer cells

penetrate into lymphatic
system and blood vessels

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 Malignant versus Benign Tumors
Benign tumors
generally do not
spread by
invasion or
metastasis
Malignant
tumors are
capable of
spreading by
invasion and
metastasis
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 What causes Cancer?
Cancer is caused by
alterations or mutations in
the genetic code
Can be induced in somatic
cells by:
Carcinogenic
chemicals
Radiation
Some viruses
Heredity - 5%
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 Oncogenes

Cellcycle

Apoptosis TumorSuppressor

Angiogenesis Inv.andMets

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HanahanandWeinberg,Cell100:57,2000
 What is the molecular basis of
cancer?
Cancer is a genetic disease.
Mutations in genes result in
altered proteins
During cell division
External agents
Random event
Most cancers result from
mutations in somatic cells
Some cancers are caused by
mutations in germline cells
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 Theories of cancer genesis
Standard Dogma
Proto-oncogenes (Ras melanoma)
Tumor suppressor genes (p53
various cancers)
Modified Dogma
Mutation in a DNA repair gene
leads to the accumulation of
unrepaired mutations (xeroderma
pigmentosum)
Early-Instability Theory
Master genes required for
adequate cell reproduction are
disabled, resulting in aneuploidy
(Philadelphia chromosome)
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 CANCER AND GENETICS
Cancer: genome disease
Causes of genomic changes
Effects of genomic changes
Revolution in cancer treatment: Smart Bullets Period

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CANCER: GENOME DISEASE

Loss of DNA
Gain of DNA
Changes in nucleotides
Epigenetic effects

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Signs for Genomic Changes
in Cancer
Changes in chromosome numbers
- Aneuploidy
Chromosomal changes
- Increase in DNA copy number -15
different region
- Loss in chromosomal -200.000 regions
Micro changes
- Microsatellite changes
Mikrosatellite - 100.000
- Nucleotide changes
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 Chromosomal changes in the genome of cancer
cells: tip of the iceberg
Reciprocal Ring
Deletion Duplication
translocation Chromosome

Terminal Insertion Inversion Robertsonian Isochromosomes

Deletion Translocation

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http://www.tokyo-med.ac.jp/genet/cai-e.htm
 Nucleotide changes in the genome of cancer
cells: unseen site of the iceberg
Nucleotide Nucleotide Nucleotide
Deletions Insertions Substitutions

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http://www.tokyo-med.ac.jp/genet/cai-e.htm
DNA Loss in cancer cells

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DNA Loss in cancer cells: beyond coincidence ...
Early Brain Tumor Advance Brain Tumor
(Astrocytoma Stage II) Glioblastoma Multiform (Stage IV)

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Chromosomal loss:

Mostly, it is a sign
for the loss of a
tumor suppressor
gene

CDKN2
locus

PTEN
locus

RB1
locus
???
locus
p53
locus

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Cancer: Genome Disease

Epigenetic effects

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Genetic and Epigenetic Silencing of Tumor Suppressor Genes

Plass - 2002 28
THE CAUSES OF GENOMIC CHANGES IN CANCER

UV

Carcinogenic Replication Errors

chemicals
Radiation

Normal cell Viruses

Damaged DNA

Rearrangements
Point mutations (translocation, deletions,
amplifications)
Alters DNA of genes controlling cell proliferation.
(Proliferation becomes abnormal)

Cancer cell

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THE CAUSES OF GENOMIC CHANGES IN CANCER:
Somatic Changes

Hasar Hasar
Kanser Riski areti
Etken Tr Etkeni

Mortesi Inlar Deri Ka., Melanoma P53 (CC-TT)

Fiziksel
Radyasyon Tiroid Ka., Lsemi Translokasyon
Benzopren Akcier Ka. p53 (G-T)
Kimyasal Aflatoksin Karacier Ka. p53 (249 G-T)
Oksidatif Stres Yallk Kanserleri P53 (C-T)
Virus DNA
Biyolojik HBV Karacier Ka.
ntegrasyonu

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THE CAUSES OF GENOMIC CHANGES IN CANCER:
Hereditary Predisposition
Genes Disease Function Inheretance Cancer Risk
FA Genes F-A DNA Damage respose ? OR Lsemi
XP Genes NER Type
X-P OR Skin Ca.
DNA Repair
BLM Bloom DNA Helicase ? OR Various cancers
WRN Werner DNA Helicase ? OR Sarcoma
Rothmund-
RECQ4 Thomson
DNA Helicase OR Sarcoma

OD Colon,
MLH1, MSH2, MMR Endometrium Ca.
PMS1, PMS2 DNA Repair
OR Lsemi, NF1
OD Breast, Ovary,
BRCA1, BRCA2 DNA Repair Prostate, Pancreas
Ca
OR Lymphoma,
A-T Leukemia
ATM DNA Damage sense ?
OD Breast Ca. ?
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p53 Li-Fraumeni DNA Damage sense OD Various cancers
 CANCER AND
GENETICS
Approximately 90-95% of all cancers
are sporadic.

5-10% are inherited.

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 GENES PLAYING ROLE IN
CANCER DEVELOPMENT

Oncogenes
Tumor suppressor genes
DNA repair genes

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 What are the genes responsible for tumorigenic
cell growth?
Normal
Proto-oncogenes + Cell growth
and
Tumor suppressor genes
- proliferation

Cancer
Mutated or activated ++
oncogenes Malignant
transformation
Loss or mutation of
Tumor suppressor genes

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 ONCOGENES

Oncogenes are mutated forms

of cellular proto-oncogenes.

Proto-oncogenes code for

cellular proteins which
regulate normal cell growth
and differentiation.
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 Five types of proteins encoded by proto-
oncogenes participate in control of cell growth:

Class I: Growth Factors

Class II: Receptors for Growth Factors and Hormones

Class III: Intracellular Signal Transducers

Class IV: Nuclear Transcription Factors

Class V: Cell-Cycle Control Proteins

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 Functions of Cellular Proto-Oncogenes

1. Secreted Growth Factors

2. Growth Factor Receptors

4. Nuclear
Proteins:
Transcription
3. Cytoplasmic Factors
Signal Transduction
Proteins
5. Cell Growth
Genes
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A generic signalling
pathway

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Oncogenes

proto-oncogene = ras
Oncogene = mutated
ras
Always activated
Always stimulating
proliferation

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 Amino acid substitutions in Ras family proteins
(inactivates GTPase)
amino acid position

Ras gene 12 59 61 Tumor

c-ras (H, K, N) Gly Ala Gln normal cells

H-ras Gly Ala Leu lung carcinoma

Val Ala Gln bladder carcinoma
K-ras Cys Ala Gln lung carcinoma
Arg Ala Gln lung carcinoma
Val Ala Gln colon carcinoma
N-ras Gly Ala Lys neuroblastoma
Gly Ala Arg lung carcinoma

Murine sarcoma virus

H-ras Arg Thr Gln Harvey strain

K-ras Ser Thr Gln Kirsten strain
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Activation mechanisms of proto-oncogenes

proto-oncogene --> oncogene

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CHROMOSOMAL REARRANGEMENTS OR TRANSLOCATIONS
Neoplasm Translocation Proto-oncogene

Burkitt lymphoma t(8;14) 80% of cases c-myc1

t(8;22) 15% of cases
t(2;8) 5% of cases

Chronic myelogenous t(9;22) 90-95% of cases bcr-abl2

leukemia

Acute lymphocytic t(9;22) 10-15% of cases bcr-abl2

Leukemia

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c-myc is translocated to the IgG locus, which results in its activated expression

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bcr-abl fusion protein is produced, which results in a constitutively active abl kinase

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 GENE AMPLIFICATION
Oncogene Amplification Source of tumor

c-myc ~20-fold leukemia and lung carcinoma

N-myc 5-1,000-fold neuroblastoma

retinoblastoma

L-myc 10-20-fold small-cell lung cancer

c-abl ~5-fold chronic myoloid leukemia

c-myb 5-10-fold acute myeloid leukemia

colon carcinoma

c-erbB ~30-fold epidermoid carcinoma

K-ras 4-20-fold colon carcinoma

30-60-fold adrenocortical carcinoma

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 Oncogenes are usually dominant
(gain of function)

cellular proto-oncogenes that have been mutated

(and activated)

cellular proto-oncogenes that have been captured by

retroviruses and have been mutated in the process
(and activated)

virus-specific genes that behave like cellular proto-

oncogenes that have been mutated to oncogenes (i.e.,
activated)

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 The result:
Overproduction of growth factors
Flooding of the cell with
replication signals
Uncontrolled stimulation in the
intermediary pathways
Cell growth by elevated levels of
transcription factors

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 Tumor suppressor genes
Normal function - inhibit cell
proliferation
Absence/inactivation of
inhibitor --> cancer
Both gene copies must be
defective

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KNUDSON TWO HIT HYPOTHESIS IN FAMILIAL CASES

Familial RB (%30)

RB rb
Normal cells

RB rb RB rb
LOH Inactivation of a tumor suppressor
gene requires two mutations, inherited
mutation and somatic mutation.
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Tumor cells Normal cells
KNUDSON TWO HIT HYPOTHESIS IN SPORADIC CASES

Normal RB RB
Cells

RB RB RB
LOH

Inactivation of a tumor
suppressor gene
RB requires two somatic
Tumor cells mutations.
Mutation
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 TUMOR SUPPRESSOR GENES
Disorders in which gene is affected

Gene (locus) Function Familial Sporadic

DCC (18q) cell surface unknown colorectal

interactions cancer

WT1 (11p) transcription Wilms tumor lung cancer

Rb1 (13q) transcription retinoblastoma small-cell lung

carcinoma

p53 (17p) transcription Li-Fraumeni breast, colon,

syndrome & lung cancer

BRCA1(17q) transcriptional breast cancer breast/ovarian

tumors
BRCA2 (13q) regulator/DNA repair
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 CELL CYCLE

Daugther cell

Mitosis Gateway

Growth
Factors

S
CELL CYCLE
DNA replication

Cell cycle
inhibitors

Control Point

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 Rb gene
Rb protein controls cell cycle moving past G1 checkpoint
Rb protein binds regulatory transcription factor E2F
E2F required for synthesis of replication enzymes
E2F - Rb bound = no transcription/replication
Growth factor --> Ras pathway
--> G1Cdk-cyclin synthesized
Active G1 Cdk-cyclin kinase phosphorylates Rb
Phosphorylated Rb cannot bind E2F --> S phase
Disruption/deletion of Rb gene
Inactivation of Rb protein
--> uncontrolled cell proliferation --> cancer

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 p53
Phosphyorylated p53 activates transcription of
p21 gene
p21 Cdk inhibitor (binds Cdk-cyclin complex -->
inhibits kinase activity)
Cell cycle arrested to allow
DNA to be repaired
If damage cannot be repaired
--> cell death (apoptosis)

Disruption/deletion of p53 gene

Inactivation of p53 protein
--> uncorrected DNA damage
--> uncontrolled cell proliferation --> cancer

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 DNA REPAIR GENES

These are genes that ensure each strand of genetic

information is accurately copied during cell division of the
cell cycle.

Mutations in DNA repair genes lead to an increase in the

frequency of mutations in other genes, such as proto-
oncogenes and tumor suppressor genes.

i.e.Breastcancersusceptibilitygenes(BRCA1andBRCA2)
Hereditarynonpolyposiscoloncancersusceptibilitygenes
(MSH2, MLH1, PMS1, PMS2) have DNA repair functions.
Their mutation will cause tumorigenesis.

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Molecular
mechanisms of
DNA double
strand break BRCA1/2
repair

Van Gent et al, 2001

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IMPORTANCE OF DNA REPAIR

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 Tumor Progression
Multiple
Cellular
mutations lead to colon
cancer
Genetic changes --> tumor
changes

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Revolution in cancer treatment:
Smart Bullets Period

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Summary of 30 years of research (1971-2001)

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Hanahan & Weinberg 2000
 HERCEPTIN

Bilimsel Aratrmalarn
ERCEPTNKanserle Savaa Katks

STI-571
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Translocation and Bcr-Abl fusion in CML

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STI571againstBcrAbl

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Smart bullet STI-571 lockes itself to the target molecule

STI-571
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 Thousands of Targets

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? ? ?
?
?

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HERCEPTIN
?
? ?

STI-571

? ?
?

? ?
?

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 MOLECULAR BIOLOGY & INFORMATICS

Biyoinformatik

~3.000.000.000 bp
DNA

~30.000 genes
~300.000 protein
~3.000.000 interaction 64
1 human cell
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