Carcinogenesis

Characteristics of Cancer
Disorder of altered cell differentiation and growth
 Results in neoplasia (“new growth”)

Growth is uncoordinated and relatively

autonomous
 Lacks normal regulatory controls over cell
growth and division
 Tends to increase in size and grow after
stimulus ceases or needs of organism are met
Components of Tissue Renewal
and Repair
Cell proliferation
 Process of cell division
 Inherent adaptive mechanism for replacing
body cells
Cell differentiation
 Process of specialization
 New cells acquire the structure and function
of cells they replace
Apoptosis
 Form
of programmed cell death to eliminate
unwanted cells
 Types of Stem Cells
Unipotent: give rise to one type of
differentiated cell
 Muscle satellite cell
 Epidermal stem cell

 Spermatogonium

 Basal cell of the olfactory epithelium

Oligopotent: produce small number of cells

Pluripotent: give rise to numerous cell
types
Determination and
Differentiation
 The Cell Cycle
Definition: The interval between each cell
division
Genetic information is duplicated
 Duplicated chromosomes are appropriately aligned
for distribution between two genetically identical
daughter cells
Checkpoints in cycle provide opportunities for
monitoring the accuracy of deoxyribonucleic acid
(DNA) replication
 Edited and repaired defects ensure full complement
of genetic information to each daughter cell
 Phase of the Cell Cycle
G1 (gap 1): the post mitotic phase
 DNA synthesis ceases while ribonucleic acid (RNA)
and protein synthesis and cell growth take place
S phase: DNA synthesis occurs, giving rise to
two separate sets of chromosomes, one for each
daughter cell
G2 (gap 2): the premitotic phase
 DNA synthesis ceases; RNA and protein synthesis
continues
M phase: the phase of cellular division or
mitosis
Control of Cell Cycle
 Control of Cell Cycle
The cell cycle is controlled by many proteins from
inside & outside the cell.
Intracellular cyclins and cyclin dependent kinases
(CDKs) control the checkpoints.
Extracellular proteins from other cells called Growth
Factors signal the target cell to divide.
Binding of growth factors to membrane receptor
proteins of the target cell triggers a molecular
signaling pathway - a series of proteins which allows
the cell to pass the checkpoints of the cell cycle.
Cell Cycle is controlled by
genes.
Signal Transduction
 Cell Cycle: Regulation
Cyclines: regulatory proteins, active at specific stages in
the cell cycle
 CDK: cycline dependent kinases

 CDI: CDK inhibitor

Tumor Suppressor Proteins Inhibit Cell
Division & Prevent Cancer
Tumor suppressor proteins are proteins that bind to
checkpoint proteins to stop the cell cycle & prevent cell
division.
An important function of tumor suppressor proteins is to
stop the division of mutated cells until mistakes in DNA
are repaired by enzymes.
TS proteins keep most mutations from being passed on
to daughter cells & developing into cancer.
If the genes for TS proteins mutate or are deleted
cancers may result.
Two important TS proteins are the p53 protein & the RB
protein.
Normal (interphase) cells are ‘parked’ in
G1 (G0) and will not proceed to S phase
unless induced.
eg by growth factors
• Therefore normal cells grown in tissue
culture need growth factors in order to
divide (proliferate)
Cancer cells have lost the G1
checkpoint regulation
 Cell Proliferation
Definition
 The process by which cells divide and
reproduce
Regulation
 Regulated in normal tissue, so the number of
cells actively dividing equal the number of
cells dying or being shed
Two Major Categories of Cells
Existing in Humans
Gametes (ovum and sperm)
 Haploid (containing one set of chromosomes
from one parent)
 Designed for sexual fusion forming a diploid
cell (containing both sets of chromosomes)
Somatic cell
 The diploid cell that forms the rest of the
body
Categories of Cell Types of the
Body
Well-differentiated neurons and cells of skeletal
and cardiac muscle unable to divide and
reproduce
Parent or progenitor cells that continue to divide
and reproduce
 Blood cells, skin cells, liver cells

Undifferentiated stem cells that can be triggered

to enter cell cycle and produce large numbers of
progenitor cells when needed
 Types of Tumors
Adenoma: benign tumor of glandular epithelial
tissue
Adenocarcinoma: malignant tumor of glandular
epithelial tissue
Carcinoma: malignant tumor of epithelial tissue
Osteoma: benign tumor of bone tissue
Sarcoma: malignant tumors of mesenchymal
origin
Papillomas: benign microscopic or macroscopic
fingerlike projections growing on a surface
Factors differentiating Benign
and Malignant Neoplasms
Cell characteristics
Manner of growth
Rate of growth
Potential for metastasizing or spreading
Ability to produce generalized effects
Tendency to cause tissue destruction
Capacity to cause death
 Characteristics of Benign
Neoplasms
A slow, progressive rate of growth that may
come to a standstill or regress
An expansive manner of growth
Inability to metastasize to distant sites
Composed of well-differentiated cells that
resemble the cells of the tissue of origin
 Characteristics of Malignant
Neoplasms
Tend to grow rapidly and spread widely
Have the potential to kill regardless of their
original location
Tend to compress blood vessels and outgrow
their blood supply, causing ischemia and tissue
necrosis
Rob normal tissues of essential nutrients
Liberate enzymes and toxins that destroy tumor
tissue and normal tissue
 Methods by which Cancer
Spreads
Direct invasion and extension
Seeding of cancer cells in body cavities
Metastatic spread through the blood or lymph
pathways
Factors Affecting Tumor Growth
The number of cells that are actively dividing or
moving through the cell cycle
The duration of the cell cycle
The number of cells that are being lost
compared with the number of new cells being
produced
 Carcinogenesis
Hypotheses of the Origin of Neoplasia

1. Oncogenes and Tumor Suppresor Genes

2. Viral Oncogene Hypothesis
3. Epigenetic Hypothesis
4. Failure of Immune Surveillance
1. Oncogenes and Tumor Suppresor
Genes
Genes that Control Cell Growth and Replication

Genes control cell division by cytokines.

Three classes of regulatory genes.
 Promotors – Proto-oncogenes
 Inhibitors – Cancer-suppressor genes – p53
 DNA stability genes.
Non-lethal Genetic damage lies at the
center of carcinogenesis.

Loss/damage to suppressor genes,

Duplication of promotor genes
Loss/damage of DNA stability genes.
 Gene Mutations That Cause
Cancer
Mutations in 4 types of genes cause Cancer
Proto - oncogenes: genes that code for normal proteins used in cell
division
 Growth factors

 Growth factor membrane receptors

 Signaling proteins

 ras proto- oncogene in 30% of cancers.

Tumor Suppressor genes: gene that code for proteins that help prevent
uncontrolled cell division by blocking key steps (e.g. DNA replication).
 Retinoblastoma susceptibilty (RB) gene

 p53 gene in >50% of cancers

DNA stability genes

Alterations of Specific Cellular
Functions in Cancer
Tumor Suppressor Oncogenes
Genes

Inactivation Activation

Differentiation Apoptosis/Proliferation

CANCER
 Proto-oncogenes
Oncogenes:
 Viral proteins which interact with the cellular controll
mechanisms to overcome the strict regulation of
proliferation (v-ras, v-myc, v-abl, ...)
Proto-Oncogenes:
 Cellular proteins which correspond to the viral
Oncogenes but which are strictly regulated. Mutations
in this genes could transform a cell into a tumor cell
(c-ras, c-myc, c-abl, ...).
Tumor genes: Knowledge
from Viruses
 Proto-oncogenes
TYPES OF ONCOGENES
1. Growth factors
2. Growth factors receptors
3. Intracellular signaling transduction factors
Proteins with GTPase activity
Cytoplasmic serine threonine kinases
4. DNA-binding nuclear proteins
5. Cell cycle factors
 Relationship between gene
products of proto oncogene
Growth factors eg IGF
Growth factor receptors
Eg erb-2, ret

Signal transducing
factors
Eg cytoplasmic
kinases

DNA binding
cell cycle proteins
proteins eg concerned
cyclin D with
 Proto-oncogenes
FUNCTION OF ONCOGENES
Cancers have characteristics that indicate, at cellular
level, loss of the normal function of oncogene products
consistent with a role in the control of cellular
proliferation and differentiation in the process known as
signal transduction. It is a complex multistep pathway
from the cell membrane, through the cytoplasm to the
nucleus.
Proto oncogenes have been highly conserved during
evolution, and the protein products they encode are
likely to have essential biological functions.
 Oncogenes Are Mutated
Proto-oncogenes
A cell can acquire a cancer causing oncogene from
 A virus

 A mutation in a proto-oncogene

Oncogenes still code for the proteins needed for

cell division but they cause cancer by producing
 Increased In growth factor

 Increased In growth factor receptors

 Increased in signal transduction

 Increase in activation of transcription

Increase Increased Increased
In growth Increase in
d in signal
factor activation
In growth transductio
receptors of
factor n
transcripti
on
 Cancer causing Mutations
Proto-oncogenes form oncogenes by
• being misplaced (e.g. by translocation) to a site where
the gene is continually expressed resulting in
overproduction of a protein that stimulates cell division
(e.g. in CML*)
• By mutating to a form that is over expressed.

Mutations in Tumor Suppressor genes cause cancer by

inactivating the genes.
 Tumor-suppressor genes
BIOLOGICAL FUNCTIONS OF TUMOR
SUPPRESSOR GENES
2. Growth Inhibitors (e.g., TGF-β; glucocortocoids)
3. Growth Inhibitor Receptors
4. Signal Transduction Protein Inhibitors
5. Transcription Factors of Growth Inhibitors
 Tumor-suppressor genes
Geneproducts which are normaly responsible for
negative controll of transcription and proliferation
Examples:
 pRb inhibits transcription factors of the E2F-family,
which are needed to get into the S-Phase of the cell
cycle (Restriction Point)
 p53 induces transcription of the CDK-inhibitor (CDI)
p21 which causes a cell cycle arrest (one function)
 p53 is found upregulated in cells with a high level of
NAdamage
Rb protein
 Tumor-suppressor genes
RETINOBLASTOMA
• Retinoblastoma (Rb) is a relatively rare, highly
malignant childhood cancer of the developing retinal
cells of the eye that usually occurs before the age of 5
years.
• Rb can occur either sporadically (non-hereditary form,
ussually involve only one eye), or be familial
(hereditary form, more commonly bilateral), which is
inherited in an AD manner, and also tend to present
at an earlier age.
Retinoblastoma
 Retinoblastoma
Two hit hypothesis
 All cells in the hereditary form have one mutated copy
of the gene RB1,i.e. the mutation is in the germline.
 Retinoblastoma
Two hit hypothesis
In the non-hereditary form a mutation in RB1 gene arises
as a post-zygotic (somatic) event sometime early in
development.
 p53 Gene
p53 senses DNA damage, and induces G1 arrest
and induces DNA repair process.
Cell with un-repairable DNA is directed to
apoptosis by p53 gene.
“P53 is a guardian of the genome.
Its homozygous loss leads to accumulation of
damaged DNA may result in malignancy”
Homozygous loss of p53 is seen in virtually
every type of cancer.
Over half of human malignant cells show loss of
p53 gene by special tests.
The p53 Tumor Suppressor
Protein
The p53 tumor suppressor protein is activated when DNA
is damaged. The p53 gene is called the “guardian angel
of the genome”

P53 activates genes

for proteins that
 Prevent cell
entering S phase
 Repair DNA

 Cause apoptosis
(if DNA is
irreparable)
 DNA Stability Genes
Monitor and maintain the integrity of the
DNA.
Loss of function promotes mutations
 Detection of DNA lesions decreased
 Repair of damage decreased or improper

 Decreased apoptosis
 Routes to Genetic Instability
based on Defective DNA Repair
 Carcinogenesis
Hypotheses of the Origin of Neoplasia
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
 Carcinogenesis
Hypotheses of the Origin of Neoplasia
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
 Carcinogenesis
Hypotheses of the Origin of Neoplasia
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
 Causes of Neoplasia
Environmental causes: (Carcinogens)
Chemicals
Viruses
Radiation
Hereditary causes- Genetic defects.
Combination – common.
Obscure defects
Carcinogenesis:
Chemical Carcinogenesis:
Initiation
 DNA damage eg.Benzpyrene

Promotion
 Histologic change – eg.
Turpentine (co-carcinogens)
Malignant transformation:
 Visible tumor formation –
further DNA damage.
Chemical Carcinogenesis:
Direct Acting Carcinogens:
 Alkylating Agents: Cyclophosphamide
Procarcinogenes (needs activation)
 Polycyclinchydrocarbons – Benzpyrene
 Aromatic amines, dyes - Benzidine

 Natural products: Aflotoxin

 Others: Vinyl chloride, turpentine etc.

 Viral Oncogenesis:
Insertion of viral nucleic acids  mutation
Alterations in Oncogenes, cancer
suppressor genes and genes regulating
DNA repair resulting in up-regulation of cell
division  Carcinogenesis.
Nobel Laureates – Varmus and Bishop
 v-fes,v-sis  proto-oncogenes.
 v-sis  sis  PDGF  Brain tumours.
 Viral Oncogenesis:

Human Papilloma Virus

 Cervical neoplasia – warts, papilloma, ca cx
Epstein-Barr virus –
 Burkitts Lymphoma, Nasopharyngeal ca.
Hepatitis B & C virus
 Hepatocellular carcinoma.
 Radiation Carcinogenesis:
Ionizing radiation  dysjunction 
random fusion  mutation.

Neoplasia

Mutations

X Ray workers – Leukemia

Radio-isotopes – Thyroid carcinoma
Atomic explosion – Skin cancer, Leukemia
 Hereditary Causes:
Due to inhereted abnormal genes.
FAP – gene C5, polyposis 
Adenocarcinoma colon
Retinoblastoma – Rb gene – (C13)
Neuroblastoma – (C17)
Trisomy 21 – Down’s syndrome –
Leukemias in infants.
Clinical Manifestations of Cancer
Tissue Integrity
 Compressed and eroded blood vessels,
ulceration and necrosis, frank bleeding, and
hemorrhage
Cancer Cachexia
 Weight loss and wasting of body fat and
muscle tissue; profound weakness, anorexia,
and anemia
Paraneoplastic Syndromes
 Manifestations in sites not directly affected by
the disease
Molecular Basis of Neoplasia:
d ity
Proto-oncogene H e re

i o n
i a t
a d
R
a l
n c i c
V-O e m
C h

t h er
O

Oncogene
 Multiple Genetic Changes
Cause Cancer
Cancers result from a series of genetic changes in a cell
lineage
Some cancers begin with an inherited germ line mutation.
Some inherited cancers follow a dominant pattern, e.g.
inherited retinoblastoma caused by a mutation in the Rb
tumor supressor gene increases cancer risk 10,000 times.
More than one somatic mutation is often necessary.
Accumulation of mutations over time leads to uncontrolled
cell division.
Example: Colon cancer develops in a stepwise fashion.
 Multiple Genetic Changes
Cause Cancer
Multiple Hits and Multiple Factors
 Knudson proposed that carcinogenesis requires 2 hits
 1st event – initiation
 Carcinogen = initiator
 2nd event – promotion
 Agent = promoter
 Multiple hits occur – 5 or more
 Each hit produces a change in the genome which is
transmitted to its progeny (ie. clone)
 Lag period
 Time between exposure (first hit) and development of clinically
apparent cancer
 Altered cell shows no abnormality during lag period
Multiple Genetic Changes
Cause Cancer
 Multiple Genetic Changes
Cause Cancer
1. DNA of a normal cell
 Thispiece of DNA is an exact copy of the DNA from which
it came. When the parent cell divided to create two cells,
the cell's DNA also divided, creating two identical copies of
the original DNA.
2. Mutation of DNA
This DNA has suffered a mutation, either through mis-
copying (when its parent cell divided), or through the
damaging effects of exposure to radiation or a chemical
carcinogen.
3. Genetically altered cell
Body cells replicate through mitosis, they respond to their surrounding
cells and replicate only to replace other cells. Sometimes a genetic
mutation will cause a cell and its descendants to reproduce even
though replacement cells are not needed.
The DNA of the cell highlighted above has a mutation that causes the
cell to replicate even though this tissue doesn't need replacement
cells at this time or at this place.
4. Spread and second mutation
The genetically altered cells have, over time, reproduced unchecked,
crowding out the surrounding normal cells. The growth may contain
one million cells and be the size of a pinhead. At this point the cells
continue to look the same as the surrounding healthy cells.
After about a million divisions, there's a good chance that one of the
new cells will have mutated further. This cell, now carrying two mutant
genes, could have an altered appearance and be even more prone to
reproduce unchecked.
5. Third mutation
Not all mutations that lead to cancerous cells result in the cells
reproducing at a faster, more uncontrolled rate. For example, a
mutation may simply cause a cell to keep from self-destructing. All
normal cells have surveillance mechanisms that look for damage
or for problems with their own control systems. If such problems
are found, the cell destroys itself. Over time and after many cell
divisions, a third mutation may arise. If the mutation gives the cell
some further advantage, that cell will grow more vigorously than its
predecessors and thus speed up the growth of the tumour.
6. Fourth mutation
The new type of cells grow rapidly, allowing for more opportunities for
mutations. The next mutation paves the way for the development of
an even more aggressive cancer.

At this point the tumour is still contained.

7. Breaking through the membrane
The newer, wilder cells created by another mutation are able to push their
way through the epithelial tissue's basement membrane, which is a
meshwork of protein that normally creates a barrier. The invasive cells
in this tumour are no longer contained.

At this point the cancer is still too small to be detected.

8. Angiogenesis
Often during the development of earlier stages of the tumour, or
perhaps by the time the tumour has broken through the basement
membrane (as pictured above), angiogenesis takes place.
Angiogenesis is the recruitment of blood vessels from the network of
neighbouring vessels.
Without blood and the nutrients it carries, a tumour would be unable to
continue growing. With the new blood supply, however, the growth of
the tumour accelerates; it soon contains thousand million cells and,
now the size of a small grape, is large enough to be detected as a
lump
9.Invasion and dispersal
The tumour has now invaded the tissue beyond the basement
membrane.

Individual cells from the tumour enter into the network of newly formed
blood vessels, using these vessels as highways by which they can
move to other parts of the body. A tumour as small as a gram can
send out a million tumour cells into blood vessels a day.
10. Tumour cells travel – metastasis
What makes most tumours
so lethal is their ability to
metastasize -- that is,
establish new tumour sites at
other locations throughout
the body.
Secondary tumours.

Metastasis is now underway,

as tumour cells from the
original cancer growth travel
throughout the body. Most of
these cells will die soon after
entering the blood or lymph
circulation.
11. Metastasis
To form a secondary tumour, a tumour cell needs to leave the
vessel system and invade tissue. The cell must attach itself to a
vessel's wall. Once this is done, it can work its way through the
vessel and enter the tissue.
Although perhaps less than one in 10,000 tumour cells will
survive long enough to establish a new tumour site, a few
survivors can escape and initiate new colonies of the cancer.
 Multi-step Theory

Initiation
Promotion
Progression
 Initiation
An initiated cell is one in which a chemical
carcinogen has interacted with DNA to produce
a mutation, often a single base alteration, in the
genome.
An initiated cell is not a tumor cell because it has
not yet acquired autonomy of growth.
The DNA alteration may remain undetected
throughout the life of the organism unless further
events stimulate development of a tumor.
 Tumor Promotion
In general, tumor promotion can be viewed as
the clonal expansion of an initiated cell via
altered gene expression that gives the cell a
selective growth advantage.
Tumor promoters cause cells to proliferate but
not to terminally differentiate, resulting in
proliferation of preneoplastic cells (benign
lesions).
Unlike initiators, most promoters do not bind
covalently to DNA and usually do not cause
mutations.
 Tumor Progression
Tumor progression describes the process
whereby tumors acquire the ability to grow,
invade local tissue and establish distant
metastases.
Increased genetic instability and karyotypic
alterations are hallmarks of progression.
Inherited or acquired mutations in genes such as
p53 or DNA mismatch repair can increase the
rate of mutation in other genes (mutator
phenotype) and, therefore, promote the tumor
progression.
 Phenotypic characteristics of
cancer cells
Immortalization
Transformation
Loss of contact growth inhibition
Autonomy of proliferation
Avoidance of apoptosis
Aberrant differentiation
Induction of angiogenesis
Overview of Carcinogenesis