ONCOGENES

CANCER
Characterized by uncontrolled
cell proliferation
Arises from irreversible genetic
damage to cells DNA, block in
normal process of
differentiation, or block in
apoptosis
Term cancer : Hippocrates (400
BC)
Observation: Veins radiating

Introduction to Genes:
Genes are carried on chromosomes and are the
basic physical and functional units of heredity .
A genome is the full set of genes in each cell
of an organism.
Form the basic unit of heredity and Encode how
to make a protein.
i.e. DNARNA proteins
Proteins carry out most of lifes function and if
altered causes a number of diseases.
When genes are altered so that the encoded
proteins are unable to carry out their normal

The cell cycle

alternate between dividing and resting or not divi

Gap 1; metabolic activity and cell growth

G0
Mitosis

1 hour of 16 hour
cell cycle

(resting phase)

DNA synthesis

Gap 2; metabolic activity and cell growth

main checkpoints in the cell c

01 Nobel Prize was awarded to 3 scientists who studied

genes that regulate the cell cycle
1. Is cell the correct size?
1.
Is DNA damaged?
3.

2. Is DNA fully replicated?

Is DNA damage repaired?

2.

3. Have spindle fibers formed?

Have they attached to
chromosomes correctly?

Progressive
Acquisition of
Neoplastic Features

Hallmarks of Cancer Cells

Self-sustained
replication
Longer survival
Genetic instability
Capable of
inducing
neoangiogenesis
Capable of
invasion and
metastasis

Apoptosis downregulation
Lack of response
to inhibitory
factors
Self-sustained
proliferation

Hallmarks of Cancer Cells

Self-maintained
replication
Longer survival
Genetic instability
Capable of
inducing
neoangiogenesis
Capable of
invasion and
metastasis

Apoptosis
downregulation
Telomerase
reactivation

Hallmarks of Cancer Cells

Self-maintained
replication
Longer survival
Genetic instability
Capable of
inducing
neoangiogenesis
Capable of
invasion and
metastasis

Cooperative
genetic
damage
Mutagenic
agents
Defective
repair systems

Hallmarks of Cancer Cells

Self-maintained
replication
Longer survival
Genetic instability
Capable of
inducing
neoangiogenesis
Capable of
invasion and
metastasis

Cancer Cells Evade Two

Safety Mechanisms
Built into the Cell Cycle
1. Once p53 is inactivated, cells with DNA
damage dont arrest from G1 and
dont undergo apoptosis.
2. Telomerase enzyme is activated,
avoiding the limit to cell divisions
imposed by telomere shortening.

Hypotheses of the
Origin of
Neoplasia
1. Oncogenes and Tumor Suppressor
Genes
2. Viral Oncogene Hypothesis
3. Epigenetic Hypothesis
4. Failure of Immune Surveillance

Origin of Neoplasia
two general types
Monoclonal
Initial neoplastic change affects a
single cell
Field origin
Carcinogen acts on large number
of cells producing field of
potentially neoplastic cells

 Oncogene : an altered form

(allele) of a normal cellular gene
(proto-oncogene).
Operationally defined as a
regulatory gene with dominant
transforming properties.
Tumor suppressor genes:
recessive genes that restrain cell
proliferation.
First discovered through their
association with specific

Alterations of Specific
CellularDNA
Functions
in
Repair
Cancer
Tumor Suppressor
Oncogenes
Genes
Activation
Inactivation

Proliferation

CANCER

Types Of Oncogenes
Two main types :
Viral oncogene: gene from the
retrovirus itself
Non-Viral oncogene (Cellular
oncogene): genes derived from the
genes of the host cell that are in an
inactive form usually. Occasionally if
the gene incorporates with the viral
genome will form a highly oncogenic
virus.

Functions Of Proto-oncogenes

 Oncogene causes cancer by

affecting:
1. Cell Proliferation: (example; Ras, Raf,
EGF)
2. Cell differentiation (example, PML/RAR
that inhibits the differentiation of
promyelocyte to granulocyte which will
maintain the cell in its active proliferate
state)
3. Cell Survival (example; Pl-3/AKT which
will activate BCL-2
inhibit
Apoptosis & maintain cell survival.

Oncogenes/Proto-oncogenes

Cyclin D1 and Cyclin E are proto-oncogenes

Often amplified or over expressed due to
other mutations (e.g. translocation) in many
cancers
Cyclin D1 allows for DNA replication (S phase)
Over expression seems to contribute to cells
progression from G0 phase and begin division

1 ) Oncogenes and Tumor

Suppressor
Genes
Proto-oncogenes

2) Viral Oncogene
Hypothesis
RNA Retrovirus produces DNA
provirus
DNA provirus containing viral
oncogene (v-onc) is introduced, or
DNA provirus without v-onc is
inserted adjacent to c-onc in host
cell DNA
RNA viruses is thought to have
acquired v-onc sequence by
recombinant mechanism from
animal cells
DNA virus

V-oncs Are Mutated Protooncogenes

SIGNAL
TRANSDUCTION
PATHWAY AS A
SOURCE OF
PROTOONCOGENES

Ras
Growth
Pathway

factor binds
receptor
Receptor
exchanges
GTP for GDP
on Ras
Ras
activated
RasRafM

V-erbB Expresses a Truncated

EGF-Receptor Which Is ALWAYS
on, regardless of [EGF]

MECHANISMS OF
CONVERSION OF
PROTOONCOGENE INTO
ONCOGENE

1) Point Mutation

2)Gene Amplification

How Cellular
Oncogenes Arise

35

3)Chromosomal Translocation

4)Local DNA Rearrangements

5)Insertional Mutagenesis
Occurs during viral DNA integration
Eg: Avian leukosis virus- integrate
within c-myc oncogene
Exon 1-unknown function
Exon 2 & 3 encode MYC protein
ALV integrates between exon 1 & 2

Tumour Suppressor Genes

Tumour Suppressor genes: are genes that act to
inhibit cell proliferation and tumour development.
If Tumor Suppresor Gene was
Mutated

OR

Inactivated

It will lead to cell transformation

Categories of tumor
suppressor genes
Caretaker genes:
Maintain the integrity of the genome
by repairing DNA damage
Gatekeeper genes:
Inhibit the proliferation or promote
the death of cells with damaged DNA

Tumor suppressor genes: functional categories and tumor

association
Category

Function

Tumor
susceptibility if
germ line
mutation

Comments

Gatekeep p53
ers

Transcription
factor

Li-Fraumeni
syndrome

Also mutated in
>50% of human
cancers

Rb1

Transcription
al regulator

Familial
retinoblastoma

Often mutated in
other cancers

APC

Regulates catenin
function

Familial
adenomatus
polyposis

Often mutated in
sporadic
colorectal cancers

BRCA
1

DNA repair

Breast and
ovarian cancer

Rarely mutated in
sporadic breast
cancers

BRCA
2

DNA repair

Breast
cancer(female
and male)

MSH2

DNA

Hereditary non-

Caretake
rs

Gene

Mutation permits

Retinoblastoma (Rb) gene

First phenotypic cancer suppressor
gene to be discovered
Responsible for retinoblastoma, a
malignant tumor of retina, a rare
childhood tumor
60% are sporadic, remaining ones are
familial

Two-hit
Hypothesis

Hereditar
y
Mutation

Nonhereditar
y
Mutation

Regulation of cell cycle

P16
P
Cdk4/cyclin D

Rb

Rb
Rb inactive

G1

G2

Cell Cycle Blocked

G1
S
M

G2

Cell Cycle Proceeds

p53 Gene
Situated at the short arm of the chromosome 17
Mutated in most of the cancer cases
Normal functions p53
Repair of damaged DNA before S-phase in the
cycle by arresting the cell cycle in G1 until the
damage is repaired
Apoptotic cell death if there is extensive DNA
damage.

p53 Gene
P53 level raise in cells with sustained cell
damage, until the damage is repaired or
cell undergoes apoptosis
Prevents propagation of possibly mutated
cells
Called the guardian of the genome

p53 Gene
P53 can lose its function by:
Non-sense mutation or mis-sense
mutation
Complex of normal p53 and mutant
p53 inactivating the function of
normal allele
Binding of normal p53 to viral

Role of p53 in cells with

damaged DNA

Structure of p53

QuickTime and a
TIFF (Uncompressed) decompressor
are needed to see this picture.

Familial cancer
syndromes involving
DNA repair enzymes
Nucleotide excision repair (NER) genes:
Xeroderma pigmentosum
. In NER-deficient cells non-repaired
dimers
lead to missense mutations during DNA
replication
Activating oncogene mutations
Inactivating tumor suppressor gene mutations

Hereditary Non-Polyposis
Colon
Carcinoma
Autosomal dominant
inheritance
Penetrance ~80%
Genes belong to
DNA mismatch
repair (MMR) family
Tumor site in proximal
colon predominates
Extracolonic cancers:
endometrium, ovary,
stomach, urinary tract,
small bowel, bile ducts,

HNPCC Results From Failure

of Mismatch Repair (MMR)
Genes
Base pair
mismatch

Normal DNA
repair

TCGAC

AGCTG

T C TA C
AGCTG
Defective DNA
repair (MMR+)

T C TA C
AGCTG

TC

TAC

AG

ATG

Mismatch Repair Failure

Leads
to Microsatellite
Instability (MSI)
Normal

Microsatellite
instability

Addition of
nucleotide repeats

Contribution of Gene
Mutations
to HNPCC
Families
Sporadic
Familial
Unknown
~30%

Rare CRC
syndrom
es
FAP

MSH2
~30%

HNPCC

Liu B et al. Nat Med 2:169, 1996

MSH6
(rare)

PMS1
(rare)
PMS2
(rare)

MLH1
~30%

Microsatellite Instability (MSI)

10%15% of sporadic tumors have MSI

95% of HNPCC tumors have MSI at
multiple loci
Routine MSI assays soon available
Normal

Electrophoresis gel

MSI tumor

Li-Fraumeni syndrome
Refers to the inherited predisposition
to develop cancers in many organs
owing to germ line mutations of p53
Affected individuals Carry germ line
mutation in one p53 allele, but tumors
display mutation at both alleles
Another example of two-hit hypothesis

Other tumor
suppressor genes

APC Gene
Implicated in familial adenomatous polyposis
coli and most sporadic colorectal cancers
APC binds to and inhibits the function of
catenin
-catenin activates certain transcription factors
that activates several genes including myc and
cyclin D
Mutant APC is unable bind -catenin to down
regulate its activity

Cancers Usually Result

from a Series of
Mutations in a Single
Colon Cancer: Cell
oncogene

oncogene

Tumor suppressors

WT-1 gene
Is deleted in hereditary Wilms
tumor(WT)
It codes for a DNA-binding protein that
represses transcription of PDGF,IGF-I
and abl2, which promotes growth
Loss of WT-1 gene expression also
occur in many breast cancers

NF-1 gene
Germ line mutation in type 1
neurofibromatosis(NF)
Encode neurofibromin, a negative
regulator of ras
Inactivation of NF-1 permits
unopposed ras, thereby promotes cell
growth

P15 and p16 genes

Inactivation identified primarily in
breast, pancreas and prostate tumors.
The gene products are cdk inhibitors
and serve as the negative regulators
of the cell cycle

BRCA1 and BRCA2 genes

Breast(BR) cancer(CA) susceptibility
genes, also incriminated in some ovarian
cancers
Involved in G1 check point
Block entry of cell into S phase, particularly
by inducing CDK inhibitor p21
Promote DNA repair by binding to RAD51

PTEN gene
Termed phosphatase and tensin homologue
Mutated in most prostate cancers and many
glioma and thyroid cancers
The gene product suppresses tumor growth
by antagonising tyrosine kinases
Regulates invasion and metastasis
Germ line mutation responsible for Cowden
syndrome
Multiple hamartoma
Increased risk of cancers of the breast,
thyroid and endometrium

RET
RETis an abbreviation for "rearranged during transfection",
as theDNA sequenceof thisgenewas originally found to
be rearranged within afibroblastcell line following its
transfectionwith DNA taken from humanlymphomacells.
Gene on chromosome 10 q 11.2
Encodes a receptor tyrosine kinase
Required for maturation of nervous system and kidney
morphogenesis

RET: Role in Normal

Development
RET deficient mice :
Die shortly after birth
Lack the Enteric Nervous System
Display Renal Agenesis (no Kidney)
Have Fewer Thyroid C Cells

Cys
Cys

GFR 1

Extracellular Domain

RET

GDNF

Cys
Cys

Cell Membrane
P
P

RAS-RAF Pathway

Intracellular Domain

PI3 Kinase Pathway

RET Proto-Oncogene in
MEN Type 2
Germline point mutations of RET are
responsible for all Forms of MEN Type
2
The mutations affect either the
extracellular or the intracellular
tyrosine kinase domain of RET
receptor.

Cys
Cys

GFR 1

RET

Extracellular Domain

Arg
Cys

Cell Membrane
P

P
P

Intracellular Domain

Over Active Signalling to Cells

Uncontrolled
Cell Proliferation

GENE Therapy

What is Gene
It is a technique for
Therapy
correcting defective

genes that are

responsible for disease
development
There are four
approaches:
1. A normal gene inserted
to compensate for a
nonfunctional gene.
2. An abnormal gene
traded for a normal
gene
3. An abnormal gene

How Does Gene

Therapy Work?

Why Viruses?

Viruses through the time of evolution have

evolved to infect the cells with great
specificity.
Viruses tend to be very efficient at
transfecting their own DNA into the host
cell genome.
This allows them to produce new viral
Types Of
particles
at Viruses
the period of synthesis of the
Retrovirus
cell

Lentiviruses
Poxviruses
Herpes Viruses
Adenovirus

Adenovirus

Some non virus mediated gene

The simplest method
delivery system:
in the direct
introduction of
therapeutic DNA into
target cells. like
microinjection &
gene gun method.

Introduction of the
foreign DNA into cell
by use liposome is
known as lipofection.
Introduction of DNA
into cell by exposing
a very brief period of
high voltage electric

Problems with Gene

Short Lived:
Therapy:
Hard to rapidly
integrate therapeutic DNA

into genome and rapidly dividing nature of

cells prevent gene therapy from long time.
Immune Response:
new things introduced leads to immune
response.
increased response when a repeat offender
enters.
Viral Vectors:
patient could have toxic, immune,
inflammatory response.
also may cause disease once inside.
Multigene Disorders:
Heart disease, high blood pressure,

Future of gene
therapy:
Can lead to the cure of most harmful
diseases like cancer, aids.
The efficient gene therapy method i.e. stem
cells gene therapy can control to heredity of
diseases.
May lead to a progress in Drug Designing.
Can Lead to cure of certain CNS(Central
Nervous

References
De Vita, Hellman and Rosenbergs
CANCER Principles and Practices of
Oncology, 8th Edition.
Schwartzs Principles of Surgery, 9th
Edition.
Sabiston textbook of Surgery, 18th
Edition.
Bailey and Loves Short Practice of
Surgery, 25th Edition.
Surgical Clinics of North America, Aug
2008.