ONCOGENES

Presented by,
Tina K.J.
2nd Sem MSc Biotechnology
CUSAT
 CANCER

Characterized by uncontrolled cell proliferation

Arises from irreversible genetic damage to cell’s
DNA, block in normal process of differentiation,
or block in apoptosis

Term ‘cancer’ : Hippocrates (400 BC)

Observation: Veins radiating from breast cancer

resembled legs of crab, hence karkinoma in Greek
& cancer in Latin
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 retroviruses [ v-oncogene ] from the Avian
Sarcoma Virus and was called src.
What Kinds of Genes are Mutated in
Cancer?

Oncogenes : positively regulate the cell cycle

(move it forward) - dominant mutations in proto-
oncogenes cause excess cell proliferation
Tumor Suppressors : negatively regulate the cell
cycle - recessive mutations in these genes
also cause excess cell proliferation, but they
generally need homozygosity for phenotypic
expression
 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.
CELL CYCLE
Regulators of Cell Cycle
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 cell’s
progression from G0 phase and begin division
 APOPTOSIS
Involves proteases called caspases
Regulated by Bcl2 and BAX
BAX homodimer promotes apoptosis, Bcl2
homodimer blocks apoptosis
Some cancer cells overproduce Bcl2 & are resistant to
some chemotherapies & radiation treatment
Proteins involved in cell cycle checkpoints regulate
pathway
Control of Apoptosis
Hypotheses of the Origin of
Neoplasia
1. Oncogenes and Tumor Suppresor
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
Two-hit Hypothesis
 1 ) Oncogenes and Tumor Suppresor
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
Do not contain viral oncogenes

Act by blocking suppressor gene products

Examples – HPV, EBV,HBV

3) Epigenetic Hypothesis

Changes in the regulation of gene

expression rather than in the genetic
apparatus
Pattern of gene expressions responsible
for tissue differentiation (ie. epigenetic
mechanism) are thought to be heritable
4) Failure of Immune Surveillance

Concepts:
Neoplastic changes frequently occur in cells
Altered DNA result in production of neoantigens
& tumor-associated antigens
Immune response (cytotoxic) to neoantigens as
foreign antigens
Neoplastic cells escaping recognition and
destruction become clinical cancers
 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.
Proto-oncogenes: are the form of cellular genes
that inactive normally but can incorporate with the
viral genome to produce a highly oncogenic virus
Funtions Of Proto-oncogenes
SIGNAL TRANSDUCTION
PATHWAY AS A SOURCE OF
PROTO-ONCOGENES
Ras Pathway
Growth factor binds
receptor
Receptor
exchanges GTP for
GDP on Ras
Ras activated
RasRafMek
Map
Kinasetranscription
factors genes
turned on
Myc is also a target of the RAS Pathway
V-onc’s Are Mutated Proto-oncogenes
V-erbB Expresses a Truncated EGF-Receptor
Which Is ALWAYS on, regardless of [EGF]
Receptor Tyrosine Kinases
under normal & abnormal
conditions
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

Mutation of the tumour suppressor gene
will cause cancer.
Example; deletion of Rb gene will cause retinoblastoma.
The development of retinoblastoma can be either:
Hereditary: a defective copy of Rb gene is inherited from the
affected parents.
Nonhereditary: in which 2 normal Rb genes are inherited and
develop mutation during life.

Retinoblastoma is developed if 2 somatic mutations

inactivate both copies of Rb in the same cell.
Inactivation of Tumour suppressor gene
will cause cancer!!!
If the Rb gene interact with DNA tumour virus
(SV40) it will induce cell transformation.

SV40
Functions of Tumour Suppressor gene
1. Antagonize the action of
oncogene. (ex.PTEN which converts
PIPIII to PIPII because PIPIII will activate
Pl-3/AKT which will activate BCL-2 that
will inhibit apoptosis and induce cell
transformation)
PIPII
PTEN PIPIII
PI-3
AKT

BCL-2

Inhibit apoptosis & induce

cell transformation
2. Transcription factors
Repressor transcription factors: example; WT1 is
a repressor that appears to suppress transcription
factor ( Insulin like growth factor) which will
contribute in the development of tumour.
Activator transcription factors: example; SMAD
family that are activated by TGF-β, leading to
inhibition of cell proliferation.
3. Regulate cell cycle :
Rb gene: that inhibits the cell cycle in the
G1 phase decrease cell
proliferation.
INK-4 gene: that produces P16
that inhibits cdk4/cyclin D action ( to
phosphorylate Rb gene to inactivate it’s
action)
P53: that produces P21 that has the same
action of P16 in inhibiting the action of
cdk4/cyclin D
Regulation of cell cycle

P16
P

Cdk4/cyclin D
Rb Rb

Rb inactive

G1
G1 S
S

M G2 M G2

Cell Cycle Blocked Cell Cycle Proceeds

4. Induce apoptosis:
P53 release will increase Bax
form holes in the mitochondria
release cytochrom c activate
apoptosis
 p53
Critical roles:
Prevents mutations and repairs DNA
Cell cycle arrest in G1
3 main functions in core domain:
Sequence specific transactivator in conjuntion with the N-
terminal transactivation domain
Recognize non-B forms of DNA
Remove nucleotides from ends of DNA by 3’-5’ exonuclease
activity
Also responsible for the activation of several proteins
involved in apoptosis.
 Structure of p53

QuickTime™ and a
TIFF (Uncompressed) decompressor
are needed to see this picture.
 MDM2

An E3 ubiquitin ligase
Inactivates p53
Binds specifically to the
N-terminus and inhibits
transactivation function
Also activates p53 to
proteasomal
degradation
 p16

Another tumor suppressor gene

Function in DNA damage prevention and repair

Like p53, mutation of this aids in tumor

progression
Prone to methylation
Methylated state of the protein causes
chromosome instability, and increases
mutation rates
MECHANISMS OF CONVERSION OF
PROTO-ONCOGENE INTO
ONCOGENE
1) Point Mutation
2)Gene Amplification
How Cellular Oncogenes Arise

56
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

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, sebaceous
skin tumors
HNPCC Results From Failure of Mismatch
Repair (MMR) Genes

Normal DNA TCGAC

Base pair repair
mismatch
AGCTG

T C T A C

AGCTG

Defective DNA repair T C T A C TCTAC

(MMR+)
AGCTG
AGATG
 Mismatch Repair Failure Leads
to Microsatellite Instability (MSI)

Normal

Microsatellite
instability
Addition of nucleotide
repeats
Epigenetic control of cancer
genes
Epigenetics:
Mechanisms of gene expression control that can be
passed from one cell to its offspring, that are not
reflected in changes in DNA sequence

Examples:
DNA methylation
Histone modification
Noncoding RNAs
DNA methylation

Some growth suppressing proteins are found to

be absent from cancers, but the promoter and
coding region are intact

Examples:
p16

RASSF1/NORE1
Local regions of DNA,
usually in gene
promoters (CpG rich
regions) maintain C-
methylation during
DNA replication

DNA methyltransferase
DNA Methylation

DNA methylation can be detected by DNA

sequencing after 5Me-dC deamidation to dU
(using bisulfite)

Reversal of DNA methylation (e.g. restoring

expression of tumor suppressor genes) is being
attempted using 5-aza Cytidine
Histone acetylation
Generally, histone acetylation is associated with
transcriptionally active genes

Histone acetylation is effected by Histone acetyl

transferase (HAT) and deacetylation by HDACs
HDAC inhibitors
make you
smarter too
 microRNAs
Rapidly emerging field

Certain, but complex mechanisms of gene expression control

Some miRNAs (e.g. miR15) have associations with cancer

Steroid hormone receptors in cancers
Cancers arising in hormonally-responsive tissues often retain a
hormone-responsive proliferation drive
“normal”
increased hormone sensitivity
Hormone receptors tend not to be mutated as oncogenes in
early tumor progression
Anti-hormone therapy is however effective in treating
hormonally-responsive tumors
Anti-estrogen therapy in breast cancer
Anti-androgen therapy in prostate cancer
Abnormal levels of hormones may predispose to cancer
due to increased cell replication
Breast cancer, endometrial cancer
Hormone drive may be endogenous or exogenous

Hormone
Antihormonal
Hormone-stimulated therapy
cell proliferation

Mutation acquisition,
Tumor progression Hormonally-responsive
Normal cell
cancer
PREVENTING CANCER
DO’S

DON’Ts