ONSET OF CANCER

 The combined activation of oncogenes and inactivation of

tumour suppressor genes drive the progression of cancer.
 Together, five biological phenomena may control the
progress of tumorigenesis:
- autonomous cell proliferation,
- increased ability to acquire genetic alterations due to deregulated

DNA repair,
- ability to grow in adverse conditions due to decreased apoptosis,
- capacity to invade tissues locally and to form distant metastases,
- ability to activate the formation of new blood vessels (a process
called angiogenesis).
 None alone is sufficient in itself, but cancer arises when
they interact together into a chain of coordinated events
that profoundly modifies the normal cellular pattern of
growth and development
Initiation- which involves damage to, and
then division of exposed cells such
that their growth potential is
changed irreversibly
Progression- denoting multiple rounds of cell
replication that result in the gradual
transition of an initiated cell
towards autonomous, cancerous,
growth involving ‘clonal selection’
Metastases- Ultimate spread of malignant
cells resulting in multiple tumour
sites
Carcinogenesis is a multistage process involving multiple genetic and
epigenetic events in proto-oncogenes, tumour suppressor genes and anti-
metastasis genes.
ONCOGENES & PROTOONCOGENES
 SIGNALS FOR NORMAL MITOSIS
• Normal cells growing in culture will not divide unless they are
stimulated by one or more growth factors present in the culture
medium
• The growth factor binds to its receptor, an integral membrane protein
• Binding of a growth factor to its receptor triggers a cascade of signaling
events within the cytosol. Many of these involve
• Kinases — enzymes that attach phosphate groups to other
proteins. Examples: the proteins encoded by SRC, RAF, ABL, and
the fusion protein encoded by BCR/ABL found in chronic
myelogenous leukemia (CML).
• or Molecules that turn on kinases- Example: ras. ras ras molecules
reside on the inner surface of the plasma membrane where they
serve to link receptor activation to "downstream" kinases like raf
Some tumor cells show mutations that alter forms of proteins
involved in these signal transduction. These include
the following:
Protein kinases;
G proteins;
Nuclear receptors;
Growth factors; and
Growth factor receptors.
Other tumor cells contain normal signal transduction proteins,
but in excessive amounts. Genes responsible for altering cells
to a cancerous state are called oncogenes
Genes can become oncogenes if:
 they become mutated so that their product becomes
constitutively active (that is, active all the time even in the
absence of a positive signal) or they produce their product
in excess.
 Possible causes: their promoter and/or enhancer has
become mutated.
• An ONCOGENE is a gene that
 when mutated or
 expressed at abnormally-high levels
contributes to converting a normal cell into a cancer cell
• All the oncogenic mutations are dominant; if the cell has
one normal gene (called a proto-oncogene) and one
mutated gene (the oncogene) at a pair of loci, the
abnormal product takes control.
• No single oncogene can, by itself cause cancer. It can,
however, increase the rate of mitosis of the cell in which
it finds itself. Cancer cells are cells that are engaged in
uncontrolled mitosis.
 RETROVIRAL ONCOGENES
• Viral oncogenes were first discovered in Rous Sarcoma Virus
(RSV), which infects and transforms chicken embryo fibroblasts
in culture to induce large sarcoma
• Deletion and temperature sensitive mutants of RSV were able to
show the presence of a single gene called “src”
• It encodes for a protein tyrosine kinase and is the factor
responsible for ability of RSV to induce tumours and transform
fibroblasts but was not necessary for replication of the virus
• 40 different highly oncogenic retroviruses have been identified
and each contains atleast one or more oncogenes- that are not
required for viral replication but responsible for cell
transformation
 PROTO-ONCOGENES
• The lack of involvement of retroviral oncogenes in viral
replication was an unexpected feature and therefore research
was focused on origin of retroviral oncogenes
• It was hypothesized that retroviral oncogenes were derived
from closely related genes of the normal cells
• These normal-cell genes from which the retroviral oncogenes
originated were called proto-oncogenes
• v-src refers to the viral form of the gene and c-src to the
cellular form
• Proto-oncogene is a normal gene that can become an
oncogene due to mutations or increased expression.
The resultant protein may be termed an oncoprotein
• Proto-oncogenes code for proteins that help to
regulate cell growth and differentiation (eg. src, ras, raf)

• They are often involved in signal transduction and

execution of mitogenic signals, usually through their
protein products.

• Upon activation, a proto-oncogene (or its product) becomes a

tumor-inducing agent, an oncogene

• It is estimated that fully 1% of the ~21,000 genes in the human

genome are proto-oncogenes
 GENERATION OF AN VIRAL ONCOGENE

 If an infection by retrovirus caused insertion

of the viral genome next to an oncogene
precursor (or proto-oncogene),
 Subsequent excision event may remove part
or all of the proto-oncogene, as well as the
viral genome,
 Then this faulty excision would have created
a novel viral genome containing a cellular
gene. Subsequent evolution of the virus
could change the cellular gene, creating an
oncogene.
 ACTIVATION
The proto-oncogene can become an oncogene by three basic activation
types:
(a) A point mutation within a proto-oncogene can cause a change in the

protein structure
(b) an increase in protein concentration, caused by
• an increase of protein expression (through misregulation)
• an increase of protein (mRNA) stability, prolonging its existence
and thus its activity in the cell
• a gene duplication resulting in an increased amount of protein in
the cell
(c) A chromosomal translocation causing
• an increased gene expression in the wrong cell type or at wrong times
• the expression of a constitutively active hybrid protein. This type
of aberration in a dividing stem cell in the bone marrow leads to
adult leukemia
• The expression of oncogenes can also be regulated by microRNAs
(miRNAs), and mutations in such microRNAs (known as oncomirs) can
lead to activation of oncogenes.
 Oncogenes Proto-oncogene
Transcribed under the control of Transcribed under control of
viral promoter and enhancer normal transcriptional regulatory
sequences sequences
As a result expressed at much Expressed at a level lower than the
higher levels oncogenes
Expression may occur in an Abnormal gene expression may
inappropriate cell type trigger the conversion of proto-
oncogene to oncogene that can
induce cell transformation
Oncogenes encode for proteins
that are different in structure and
function from the normal cellular
ones
Oncogenes generally differ from
their proto-oncogene by a point
mutation
 Oncogenes in human cancer
• Gene transfer assays and other experiments identified several oncogenes
in human cancers
Ras oncogene:
• One of the first to be identified was the human homolog of the ras H
oncogene of Harvey sarcoma virus
• Three closely related members of the ras family were identified – ras H,
ras K, ras N. They were known to be involved in ~25% of all human
malignancies
• ras oncogene was not present in normal cells but generated in tumour
cells as a result of mutations during tumorigenesis
• A point mutation replacing valine for glycine at position 12 converts the
ras proto-oncogene to an oncogene
• ras oncogene encodes for guanine binding proteins that functions in
transduction of mitogenic signals from variety of growth factor receptors
• Its activity is controlled by GTP (active)/ GDP (inactive) binding
• Mutation in ras gene results in decreased GTPase activity as a result ras
is constitutively activated in the GTP bound state, resulting in unregulated
cell proliferation
c-myc oncogene:
• It was one of the first characterized example of proto-
oncogene activation by chromosome translocation
• It is involved with Human Burkitt’s lymphoma and mouse
plasmacytoma, which are cancers of the B-lymphocytes
• In these cancers, there is chromosomal translocation of
genes encoding for immunoglobulins. Commonly, the
fragment from chromosome 8 is translocated to one of the
loci on either chromosome 2 ( light chain), 14 (heavy chain)
and 22 ( light chain).
• Such translocation resulted in insertion of the c-myc into an
immunoglobulin locus after moving from chromosome 8.
• Here at this new loci, the gene is expressed unregulated,
resulting in excess cell proliferation because c-myc gene
encodes a transcription factor.
• L-myc and N-myc are oncogenes
associated with lung carcinoma and
neuroblastoma.
• The activation mechanism for these is
not translocation but instead
amplification
Bcl-2:

• It was also first discovered as the product of an oncogene

in human lymphomas
• It was generated by chromosomal translocation that
results in elevated expression of Bcl-2, which blocks
apoptosis and maintains cell survival
• This discovery helped to understand the significance of
programmed cell death in development of cancer
Functions of oncogenic products:
• Uncontrolled proliferation of cancer cells:
Cancer cell produces growth factors (i.e. oncogene proteins) to
which it also responds, resulting in autocrine stimulation of the
growth factor producing cell, which drives abnormal cell
proliferation and tumour development
• Defective differentiation:
There are a number of transcription factors whose oncogenic
activity results from inhibition of cell differentiation. Thyroid
hormone and retinoic acid induce differentiation of cell types.
Mutated forms of the thyroid hormone receptor (ErbA) and
retinoic acid receptor (PML/RAR ) act as oncogene proteins.
These mutated forms interfere with action of their normal
homolog and thereby block cell differentiation and maintaining
the leukemic cells in an actively proliferating state.
• Failure to undergo apoptosis: Several oncogenes encode
proteins that act to promote cell survival. Eg. Oncogenes that
encode growth factors, growth factor receptors, signaling
proteins like ras. PI3-kinase and Akt act as oncogenes and
control Bad, a proapoptotic protein of Bcl-2 family. This is
phosphorylated by Akt and as a result inactivated. Thus, it
cannot bind Bcl-2 protein and so tumour cannot go for
apoptosis
Akt Signaling
SIGMA-ALDRICH
TUMOR SUPRESSOR GENES
 Activation of cellular oncogenes is one of the genetic
changes that occur during tumour development
 The other change involves inactivation of tumour
suppressor genes
 While oncogenes and their products are involved with
abnormal cell proliferation then tumour suppressor genes
normally inhibit cell proliferation and tumour development
 In most tumours these are lost/ inactivated allowing abnormal
proliferation of tumour cells.
 Loss of function of a tumour suppressor gene is typically a
recessive mechanism, i.e. both copies of the gene need to be
inactivated in order to switch off the corresponding function.
 The first tumour suppressor gene identified was the
retinoblastoma gene (Rb gene)
 It was stated that the inheritance of the susceptibility gene (Rb
gene) was not sufficient for occurrence of retinoblastoma (a
rare childhood eye tumor), instead it required two mutations,
which resulted in loss of both of the functional copies of the
tumor susceptibility gene (Rb tumor suppressor gene) .
 This was the two-hit hypothesis put forward by Alfred Knudsen
in 1971, which further led to the modern concept of recessive
tumour suppressor genes
 The Rb gene encodes a protein that binds and inhibits
progression of the cell cycle past the G1 restriction point,
thus fulfilling the functions of a molecular “brake” on cell division
 Most tumours develop by the loss of the normal Rb allele.
Besides it is the target of oncogenic proteins of several DNA
tumor viruses, which inactivate Rb
Tumour suppressor genes were of two types:
Gatekeepers: Their products control the gates on the pathways
of cell proliferation. Typically, gatekeeper genes are negative
regulators of the cell cycle, acting as “brakes” to control cell
division
Caretakers: their primary function is not to control the speed or
timing of cell division but rather its accuracy. Usually, involved
in DNA repair and in the control of genomic stability.
 Whereas the study of retroviruses and gene transfer
assays were the keys to the discovery of oncogenes,
tumour suppressor genes were identified through the study
of large DNA viruses (SV 40, polyoma viruses, adenoma
viruses) and the analysis of familial tumour syndromes
 This was achieved in 1989, when it emerged that the p53
gene was often mutated and/or deleted in many forms of
cancers.
 In 1991, inherited loss of p53 was found to be associated
with a rare familial syndrome of multiple cancers, the Li-
Fraumeni syndrome, in which afflicted family members
suffer vastly increased incidence of many tumour types.
 Today, about 215 families worldwide affected by this
syndrome have been described
p53 – the guardian of the genome
• The p53 gene encodes a phosphoprotein of molecular weight
53,000 daltons, which accumulates in the nucleus in
response to various forms of stress, in particular, DNA
damage.
• In this context, p53 acts as a transcriptional regulator,
increasing or decreasing the expression of several dozen
genes involved in
- cell cycle control,
- in the induction of apoptosis,
- in DNA repair and
- in differentiation control.
• Together these genes exert complex, anti-proliferative effects
 p53 acts as the “guardian of the genome

Cells subjected to Activation of p53 Cell cycle arrest

tolerable levels of
DNA-damaging
agents
temporarily removing

the cells from

the

Highly damaging Activation of p53 Induce apoptosis

proliferative pool
levels of
genotoxic
stress
Elimination of cells with potentially
oncogenic alterations
• p53 gene differs from most other tumour suppressors in its
mode of inactivation in human cancers.
• Whereas most tumour suppressors are inactivated by loss
of alleles or inactivating deletions or insertions, p53 -
target of point mutations within the portion of the gene that
encodes the DNA-binding domain of the protein
• These mutations prevent the correct folding of this protein
domain, and therefore disrupt the interactions of p53 with its
specific DNA targets. Mutation is not the only way to alter p53
protein in cancer.
• In cervical cancers, p53 gene mutations are infrequent, but
the protein is inactivated by binding of the viral protein
E6 which is produced by human papillomavirus.
• Experimental gene therapy has shown that it may be possible
to restore p53 function in cells that have lost the gene. More
recently, anti-cancer drugs were designed to specifically
target p53.