Tumor suppressor gene

A tumor suppressor gene, or antioncogene, is a gene that protects a

cell from one step on the path to cancer. When this gene mutates to
cause a loss or reduction in its function, the cell can progress to
cancer, usually in combination with other genetic changes. The loss of
these genes may be even more important than protooncogene/oncogene activation for the formation of many kinds of
human cancer cells. Tumor suppressor genes can be grouped into
categories including caretaker genes, gatekeeper genes, and
landscaper genes; the classification schemes are evolving as medicine
advances, learning from fields including molecular biology, genetics,
and epigenetics.

Two-hit hypothesis
Unlike oncogenes, tumor suppressor genes generally follow the "twohit hypothesis," which implies that both alleles that code for a
particular protein must be affected before an effect is manifested. This
is because if only one allele for the gene is damaged, the second can
still produce the correct protein. In other words, mutant tumor
suppressors' alleles are usually recessive whereas mutant oncogene
alleles are typically dominant.
The two-hit hypothesis was first proposed by A.G. Knudson for cases of
retinoblastoma.Knudson observed that the age of onset of
retinoblastoma followed 2nd order kinetics, implying that two
independent genetic events were necessary. He recognized that this
was consistent with a recessive mutation involving a single gene, but
requiring biallelic mutation. Oncogene mutations, in contrast, generally
involve a single allele because they are gain-of-function mutations.
There are exceptions to the "two-hit" rule for tumor suppressors, such
as certain mutations in the p53 gene product. p53 mutations can
function as a "dominant negative," meaning that a mutated p53
protein can prevent the function of normal protein from the unmutated allele.
Other tumor-suppressor genes that are exceptions to the "two-hit" rule
are those that exhibit haploinsufficiency, including PTCH in
medulloblastoma and NF1 in neurofibroma. An example of this is the
p27Kip1 cell-cycle inhibitor, in which mutation of a single allele causes

increased carcinogen susceptibility.

Types of Tumor Suppressor Genes

There are two main categories of Tumor Suppressor Genes (TSGs):
gatekeepers and caretakers. Gatekeepers control cell proliferation by
regulating the cell cycle whereas caretakers maintain the integrity of
cellular genetic material. Both types of TSGs are important in
preventing tumorigenesis.

Gatekeepers
Gatekeepers are genes encoding proteins that regulate cell
proliferation. They act as guards that prevent cells from passing
through cell cycle checkpoints by countering the progression of cellular
growth and encouraging apoptosis. Gatekeepers are cell-type specific
and normally prevent uncontrolled growth and potential progression of
cancer. Before considering the effects of a dysfunctional gatekeeper
gene, we must understand the carefully orchestrated regulation of the
normal cell cycle.
The cell cycle is a highly regulated process, for which two major
players are primarily responsible cyclins and cyclin-dependent
kinases (Cdks). Cyclins are a family of regulatory proteins that are
present at varying concentrations throughout the cell cycle . The
binding of cyclin to Cdk creates a cyclin-Cdk complex, which plays a
role in the progression of cells through the specific stages of the cell
cycle by regulating cell-cycle checkpoints.

Figure 6.2.1. Cyclin levels throughout the cell cycle.

Gatekeepers interact either directly or indirectly with these cyclin-Cdk

complexes. Thus, deactivation of gatekeeper genes by mechanisms
mentioned in the previoussection, allows cells to bypass the cell cycle
checkpoints, leading to unrestricted proliferation, differentiation, and
immortality of tumor cells (1). Mutation of one gatekeeper gene
predisposes an individual to cancer, while mutation of two gatekeeper
genes often leads to neoplasianew growth in excess of normal
tissue. Often, one cell type will have several gatekeeper proteins (1).

Caretakers
Caretaker genes, a second type of tumor suppressor gene, maintain
and protect the integrity of the genome and may sometimes be
referred to as "genome maintenance genes." In general, these genes
are involved in DNA repair and help prevent accumulation of
mutations. Caretaker mechanisms include DNA base excision repair,
nucleotide excision repair, prevention of oncogenic chromosomal
rearrangement, non-homologous end joining, mismatch repair and
telomere maintenance . Two examples are MLH1/MSH2, which are
involved in DNA mismatch repair, and XP-A, which is involved in the
nucleotide excision repair pathway .

A faulty protein arrising from a mutant caretaker gene can lead to

certain genomic instabilities such as microsatellites, point mutations,
or chromosomal instabilities .In humans, chromosmal instabilities,
brought about by chromosomal rearrangement errors, are the cause of
many hereditary predispositions to cancer. Mutations in caretakers
indirectly cause cancer because affected cells easily accumulates
mutations due to impaired genome maintenance. There must be two
knockouts in the caretaker gene as well as other mutations for cells to
initiate tumor formation .

Landscapers
Landscaper genes encode products that help create environments that
control cell growth. Landscapers were first discovered when lesions
were found in cells surrounding tumour tissue in juvenile polyposis
syndrome (JPS) . It was observed that the initiating lesions occured not
in the tumour cells, but rather in the surrounding stromal cells. Without
functional copies of a landscaper gene, the microenvironment may
become abnormal such that it promotes transformation of normal
epithelial cells. Predicted mechanisms involving these landscaper
genes include regulation of extracellular matrix proteins, cellular
surface markers, cellular adhesion molecules, and growth factors . One
possibility is that mutants in landscapers involved in regulating cell
membranes may trigger chemokine release, leading to unregulated
cellular proliferation. An example of a landscaper gene is PTEN,
although some evidence suggests that Rb can also function as a
landscaper .

Functions
Tumor-suppressor genes, or more precisely, the proteins they code for,
either have a dampening or repressive effect on the regulation of the
cell cycle or promote apoptosis, and sometimes do both. The functions
of tumor-suppressor proteins fall into several categories including the
following:

1. Repression of genes that are essential for the continuing of the

cell cycle. If these genes are not expressed, the cell cycle does
not continue, effectively inhibiting cell division.
2. Coupling the cell cycle to DNA damage. As long as there is
damaged DNA in the cell, it should not divide. If the damage can
be repaired, the cell cycle can continue.
3. If the damage cannot be repaired, the cell should initiate
apoptosis (programmed cell death) to remove the threat it poses
for the greater good of the organisms produced
4. Some proteins involved in cell adhesion prevent tumor cells from
dispersing, block loss of contact inhibition, and inhibit
metastasis. These proteins are known as metastasis suppressors.
5. DNA repair proteins are usually classified as tumor suppressors
as well, as mutations in their genes increase the risk of cancer,
for example mutations in HNPCC, MEN1 and BRCA. Furthermore,
increased mutation rate from decreased DNA repair leads to
increased inactivation of other tumor suppressors and activation
of oncogenes.

Examples
The first tumor-suppressor protein discovered was the Retinoblastoma
protein (pRb) in human retinoblastoma; however, recent evidence has
also implicated pRb as a tumor-survival factor.
Another important tumor suppressor is the p53 tumor-suppressor
protein encoded by the TP53 gene. Homozygous loss of p53 is found in
65% of colon cancers, 3050% of breast cancers, and 50% of lung
cancers. Mutated p53 is also involved in the pathophysiology of
leukemias, lymphomas, sarcomas, and neurogenic tumors.
Abnormalities of the p53 gene can be inherited in Li-Fraumeni
syndrome (LFS), which increases the risk of developing various types of
cancers.
PTEN acts by opposing the action of PI3K, which is essential for antiapoptotic, pro-tumorogenic Akt activation.

As costs of DNA sequencing have diminished, many cancers have now

been sequenced for the first time, revealing novel tumor suppressors.
Among the most frequently mutated genes are components of the
SWI/SNF chromatin remodeling complex, which are lost in about 20%
of tumors.
Other examples of tumor suppressors include pVHL, APC, CD95, ST5,
YPEL3, ST7, and ST14.
Example 1: RB - the retinoblastoma gene
Retinoblastoma is a cancerous tumor of the retina. It occurs in two
forms:

Familial retinoblastoma

Multiple tumors in the retinas of both eyes occurring in the first weeks
of infancy.
Familial retinoblastoma occurs when a baby inherits from one of its
parents a chromosome (number 13) that has its RB locus deleted (or
otherwise mutated). The normal Rb protein controls the cell cycle. It
integrates the signals reaching the cell to determine whether it is safe
for the cell to complete the passage from G1 of the cell cycle to
mitosis.
A random mutation of the remaining RB locus in any retinal cell
which are nondividing cells and should not enter the cell cycle
completely removes the inhibition provided by the Rb protein, and the
affected cell grows into a tumor. So, in this form of the disease, a
germline mutation plus a somatic mutation of the second allele leads
to the disease.

Sporadic retinoblastoma

A single tumor appears in one eye sometime in early childhood before

the retina is fully developed and mitosis in it ceases.
In this disease, both inherited RB genes are normal and a single cell
must be so unlucky as to suffer a somatic mutation (often a deletion) in
both in order to develop into a tumor. Such a double hit is an
exceedingly improbable event, and so only rarely will such a tumor
occur. (In both forms of the disease, the patient's life can be saved if
the tumor(s) is detected soon enough and the affected eye(s)
removed.)
Example 2: p53
The product of the tumor suppressor gene p53 is a protein of 53
kilodaltons (hence the name). (You will find that the human gene is
variously designated as P53, TP53 ["tumor protein 53"], and TRP53
["transformation-related protein 53"])
The p53 protein prevents a cell from completing the cell cycle if its
DNA is damaged or
the cell has suffered other types of damage. When the damage is
minor, p53 halts the cell cycle hence cell division until the
damage is repaired. the damage is major and cannot be repaired, p53
triggers the cell to commit suicide by apoptosis.
These functions make p53 a key player in protecting us against cancer;
that is, it is an important tumor suppressor gene. More than half of all
human lung, ovarian, and colorectal cancers harbor p53 mutations and
have no functioning p53 protein.
Mice have been cured of cancer by treating them with a peptide that
turns on production of the p53 protein in the tumor cells. However,
there may be a tradeoff involved: excess production of the p53 protein
leads to accelerated aging in mice .
Example 3: p16INK4a
The product of the tumor suppressor gene INK4a is a protein of 16
kilodaltons (hence the name).Like p53, it blocks progression through

the cell cycle in this case by inhibiting the action of the cyclindependent kinase Cdk4.
As an animal ages, its cells produce increasing amounts of p16INK4a.
This is probably a good thing in that it reduces the risk of the cell
entering uncontrolled mitosis, i.e., becoming a cancer. However, again
like p53, there is a tradeoff. As levels of p16INK4a rise in adult stem
cells and progenitor cells, their ability to reproduce and thus replace
lost or damaged tissue diminishes.
p16INK4a is not simply a reflection of an aging cell but is actively
involved in the process.
Mice expressing higher-than-normal levels of p16INK4a show earlier
replicative senescence while mice in which p16INK4a activity is
blocked continue to repair damaged tissue efficiently but run a higher
risk of getting cancer.
In mice, eliminating senescent cells (they are high in p16INK4a)
prevents (in young mice) and partially reverses (in older mice) some of
the signs of aging such as cataracts, and loss of adipose tissue and
skeletal muscle mass.
In humans, deletions and other mutations of p16INK4a are found in a
variety of cancers.