CARCINOGENESIS

The Molecular Basis of Cancer

Nonlethal genetic damage lies at the heart of

carcinogenesis
3 classes of normal regulatory genes: growth
promoting (proto-oncogenes), anti-oncogenes
(growth inhibiting / suppressor genes), apoptotic
genes (regulate programmed cell death) the
principal targets of genetic damage.
DNA repair genes, affect cell proliferation or
survival indirectly by influencing the ability of the
organism to repair non-lethal damage of other
genes.
Carcinogenesis is a multi-step process at both the
genetic and phenotypic level

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 Oncogenes: ONCOGENES
ACTIVATION
Mechanism by which protooncogenes are
transformed into oncogenes
Changes in the structure of the gene abnormal
gene product (oncoprotein)
- Point mutations
- Insertion and deletion
Changes of regulation: gene expression
enhanced / inappropriate production of the
structurally normal growth-promoting protein
- Chromosomal translocations
- Gene amplification
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 Oncogenes activation:

POINT MUTATION
ras oncogene the besat example
A very large number of human tumors carry ras
point-mutations
Mutation affect a domain critical to the GAP-
induced hydrolysis of GTP mutant ras proteins
have a reduced ability to hydrolyze GTP
Frequency:
- 90%: pancreatic adenocarcinoma
- 50%: colon and thyroid cancer
- 30%: lung adenocarcinoma & myelid leukemia
- 0%: most ovarial and breast tumors

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 Model for Action of ras gene

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 Oncogenes activation:
Insersi & Delesi
Insersi: infeksi virus
Delesi: hilangnya satu atau beberapa
basa pada susunan DNA

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 Oncogenes activation:
Chromosomal
translocations

Burkitts lymphoma

Chronic Myelogenous
Leukemia

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 Oncogenes activation:

Gene Amplification

Amplification of N-myc
gene in neuroblastoma:
double minutes / HSR
(Homogenous-staining
region)

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 Cancer suppressor genes
and apoptotic genes

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 CANCER SUPPRESSOR GENES
Misnomer
Physiologic function: regulate cell growth (not to
prevent tumor formation) apply brakes to cell
proliferation
Discovered by studying rare disease such as
retinoblastoma
Knudson Hypothysis as a paradigm for two-hit
hypothysis of oncogenesis amply substantiated by
other suppressor gene, and now can be formulated in
more precise terms, using retinoblastoma as paradigm

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 CANCER SUPPRESSOR GENES
Two-hit Hypothysis of Oncogenesis

In hereditary cases, one genetic change (first hit)

is inherited from affected parent present in all
somatic cells of the body
The second mutation (second hit) occurs in one of
many retinal cells (which already carry the first
mutation)

In sporadic case, both mutations (hits) occur

somatically within a single cell tumor
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 CANCER SUPPRESSOR GENES:

Rb gene

Paradigm of:

two hit hypothysis of

carcinogenesis

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 Role of RB as Cell-cycle Regulator

Virtually all cancers show dysregulation of the cell cycle by

affecting the four genes (red asterisk)
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 Sub-cellular location of protein
product of tumor suppressor genes

2 broad categories regarding the functions:

Molecules that regulate nuclear transcription and cell
cycle
Cell surface: TGF-receptor, E-cadherin
Under plasma mebrane: NF-1
Cytoskeleton: NF-2
Cytosol: APC/-catenin, PTEN, SMAD 2, SMAD 4
Molecules that regulate signal tranduction
Nucleus: Rb, p53, WT-1, p16(INK4a), BRCA-1,
BRCA-2

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 Selected tumor-suppressor gene involved in human neoplasm

TGF- receptor

Function: Growth inhibition

Tumors associated with somatic mutation:
Carcinoma of colon
Tumors associated with inherited
mutation: Unknown

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 Selected tumor-suppressor gene involved in human neoplasm

E-cadherin
Function:
Cell adhesion
Tumors associated with somatic
mutation:
Ca. gaster & breast
Tumors associated with inherited
mutation:
Familial gastric cancer

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 Selected tumor-suppressor gene involved in human neoplasm

NF-1
Function:
Inhibition of ras signal
transduction
Tumors associated with somatic mutation:
Schwannoma
Tumors associated with inherited mutation:
Neurofibromatosis type 1 and
sarcomas

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 Selected tumor-suppressor gene involved in human neoplasm

NF-2
Function:
Unknown
Tumors associated with somatic mutation:
Schwannoma and meningioma
Tumors associated with inherited mutation:
Neurofibromatosis type 2,
acoustic schwannoma &
meningioma

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 Selected tumor-suppressor gene involved in human neoplasm

APC
Function:
Inhibition of signal transduction
Tumors associated with somatic mutation:
Ca. of stomach, colon, pancreas;
melanoma
Tumors associated with inherited mutation:
Familial Adenomatous Polyposis
coli; colon cancer

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 Selected tumor-suppressor gene involved in human neoplasm

Rb
Function:
Regulation of cell cycle
Tumors associated with somatic mutation:
Retinoblastoma, osteosarcoma,
Ca breast, colon, lung
Tumors associated with inherited mutation:
Retinoblastoma, osteosarcoma

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 Selected tumor-suppressor gene involved in human neoplasm

p53

The guardian of the genome

Located on chromosome 17p13.1
The most common target for genetic alteration in
human tumors
A little over 50% of human tumors contain
mutation in this gene

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 Selected tumor-suppressor gene involved in human neoplasm

p53
Function:
Regulation of cell cycle & apoptosis
in response to DNA damage
Tumors associated with somatic mutation:
Ca. gaster & breast
Tumors associated with inherited mutation:
Li-Fraumeni syndrome
Multiple carcinoma and sarcoma

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 The role of
p53 in
maintaining
the integrity
of the
genome

THE
GUARDIAN
OF THE
GENOME
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 Selected tumor-suppressor gene involved in human neoplasm

WT-1
Function:
Nuclear transcription
Tumors associated with somatic mutation:
Wilms tumor
Tumors associated with inherited mutation:
Wilms tumor

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 Selected tumor-suppressor gene involved in human neoplasm

p16(INK-4a)
Function:
Regulation of cell cycle by
inhibiting CDK
Tumors associated with somatic mutation:
Pancreatic, esophageal cancer
Tumors associated with inherited mutation:
Melanoma

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 Selected tumor-suppressor gene involved in human neoplasm

BRCA-1
Function:
DNA repair
Tumors associated with somatic mutation:
Unknown
Tumors associated with inherited mutation:
Ca of female breast and ovary

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 Selected tumor-suppressor gene involved in human neoplasm

BRCA-2

Function:
DNA repair
Tumors associated with somatic mutation:
Unknown
Tumors associated with inherited mutation:
Ca of male and female breast

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 Apoptosis
(programmed cell death)

Internally programmed and coordinated death / loss of

single cells spread among healthy cells,
in a form of cell death designed to eliminate
unwanted cells,
through the serial event activities,
by a set of responsible gene product.

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 Genes that Regulate Apoptosis
bcl-2 family:
Antagonists
bcl-2 and bcl-xL
Agonists
bax, bcl-xS,
bad, bid

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 Regulation of cell death

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 Biological Mechanism

1. Signaling pathways apoptosis

initiation
2. Control and integration balance
between negative regulatory molecule
(inhibit) and positive (stimulate)
3. Common-execution phase actual
death program accomplished largely by
caspase family protease
4. Removal of death cells by phagocytosis

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 APOPTOSIS

Caspase: cystein protease, unique ability of these enzymes

to cleave after aspartic residue
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 APOPTOSIS

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 The role of apoptosis
1. Growth an development
- Embriogenesis: implantation, organogenesis,
differentiation, involution
2. Homeostatic mechanism to maintain cell population in
tissue
- hormone dependent involution
- proliferative cells population: intestinal crypt epihtel
3. Defense mechanism
- neutrophils death during an acute inflammatory
response
- cell death induced by cytotoxic T cells: in cellular
immune rejection
4. When cells are damaged by disease or noxious agent
- virus infection: hepatitis Councilman bodies
- low doses injurious stimuli: heat, radiation, anti-cancer
drugs, hypoxia
5. Aging process

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 MORPHOLOGY
Cell shrinkage
Chromatin condensation
Formation of cytoplasmic blebs and
apoptotic bodies
Phagocytosis of apoptotic cells by
adjacent healthy cells (parenchymal
cells or macrophages)
Intact plasma membrane during
apoptosis

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 MORFOLOGI ultrastructural change sequences
NECROSIS & APOPTOSIS

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 APOPTOSIS: ultrastructure feature

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 Apoptosis in skin with
immune reaction

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 Biochemical features

Protein Cleavage
Specific: protein hydrolysis involving the activation
of several members of cystein protease (caspase)
Protein cross-linking
Transglutaminase activation: apoptotic bodies
DNA breakdown
DNA pecah breakdown 50 300 kbp
Internucleosomal breakdown by endonuclease
(Ca++ & Mg++ dependent) endonucleosom
(120 200 kbp) DNA ladders
Phagocytic recognition
Apoptotic cells ekspress phosphatidylserine

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 Electrophoresis of the
DNA extractedi from cell
culture underwent
apoptosis
A. CONTROL
B. APOPTOSIS
C. NECROSIS

DNA ladders
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 Perubahan fenotipik
Sifat pertumbuhan
- lepas dari kontrol
- kegagalan maturasi
- transplantable
- immortal
Perubahan morfologik
Kariotipik
Antigenik
Deviasi metabolik
Membran sel
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 Patogenesis Kanker - resumee

The Hallmark of Cancer

Self Sufficiency in Growth Signals
Insensitivity to Growth Inhibitory
Signals
Evasion of Apoptosis
Limitless Replicative
Development of sustained
angiogenesis
Ability to Invade and Metastasize

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 The
Hallmark
of Cancer

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 The Hallmark of Cancer

Self Sufficiency in Growth Signals

Growth Factors
(PDGF, TGF-)
Growth Factor Reseptor
ERBB1 (EGF receptor), ERBB2 (HER)
Signal Transducing Protein
RAS & ABL
Nuclear Transcription Factors
MYC, MYB, JUN, FOS, REL
Cyclin and Cyclin-Dependent Kinases (CDK)

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 The Hallmark of Cancer

Insensitivity to
Growth Inhibitory Signals

RB Gene and Cell Cycle

Transforming Growth Factor
(TGF-) Pathway
Adenomatous Polyposis Coli
-Catenin Pathway
TP53 Gene: Guardian of the
Genome

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 The Hallmark of Cancer

Evasion of Apoptosis
Aktivasi onkogena
pemacu pertumbuhan

Inaktivasi gena Transformasi sel

supresor tumor

Mutasi gena
Regulasi apoptosis

Menghindari apoptosis
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 The Hallmark of Cancer

Limitless Replicative
Sel normal mempunyai kapasitas
menggandakan diri (doubling) 60-70X non-
replicative senescence (karena pemendekan
telomer terus menerus) dapat dicegah
dengan cara aktivasi enzim telomerase
aktif pada sel stem, absen pada sel somatik
aktif pada sel tumor telomer tetap panjang
replikasi terus terjadi

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 The Hallmark of Cancer

Angiogenesis terus-menerus

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 The Hallmark of Cancer

Ability to
Invade and
Metastasize

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 The Hallmark of Cancer

Ability to Invade and Metastasize

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 Patogenesis Kanker
FAKTOR-FAKTOR LAIN

Genomic Instability
Enabler to Malignancy
Molecular Basis of
Multistep Carcinogenesis

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 FAKTOR-FAKTOR LAIN
Genomic Instability Enabler to
Malignancy
Gangguan pada kemampuan reparasi DNA genom
menjadi tidak stabil contoh:
Sindroma HNPCC
Kelainan yang diturunkan: xeroderma pigmentosum
rentan terhadap sinar UV
Mutasi p53 pada beberapa kelainan, mis: sindroma
Bloom, ataxia-telangiektasia sensitif terhadap radiasi
ionisasi; Anemia Fanconi sensitif terhadap nitrogen
mustard (DNA cross-linking agent)
BRCA1 & BRCA2 pada Ca payudara

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 FAKTOR-FAKTOR LAIN

Molecular Basis of
Multistep Carcinogenesis
Karsinogenesis adalah suatu proses multistep

Berbagai macam kanker mempunyai jalur patogenesis multipel

masing-masing (tidak sama)
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 Molecular model for the evolution of
colorectal carcinoma

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 Daftar Pustaka
Robbins (1999): Pathological Basis of
Disease
Robbins (2003): Basic Pathology
Rubin (2005): Essential Pathology
Underwood (2991): General and Systemic
Pathology

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