Molecular basis of cancer:

Carcinogenesis & Regulator

genes

Ika Rahmawati Sutejo

Departement of Biochemistry
2016
 Topic
General Principles of Cancer
Definition of tumor and cancer
System of tumor nomenclature
Characteristic of benign and malignant tumors
Cancer Epidemiology
Risk Factors

Molecular Basis of Cancer from Normal Cell to Cancer Cell

Molecular Basis of Multisteps Carcinogenesis
Hallmarks of cancer
Oncogenes and cancer
Cancer suppressor genes
Apoptosis & cell cycle

Clinical Pathobiology of Cancer

Tumor antigens and tumor markers
Immunosurvaillance
Clinical Features of Tumors: Effect of tumor on host, Grading and staging of tumors,
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management
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What is cancer

Cancer : abnormal cell growth in a tissue

escape from normal control
invasive, metastasis

How does cancer occur ?

Proliferation Apoptosis

Tumor Degenerative disease
Malignancy Autoimmune disease
Multiple genetic abnormalities
Multi step 4
Carcinogenesis

5
Initiation-promotion scheme

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Inisiation & promotion

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Tumor Progression and Heterogeneity

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 HALLMARK
OF CANCER

1. Sustaining proliferative signaling

2. Evading growth suppressors
3. Avoiding immune destruction
4. Enabling replicative immortality
5. Tummor promoting
inflammation
6. Activating invasion and
metastasis
7. Inducing angiogenesis
8. Genome instability and
mutation
9. Resisting cell death
10. Deregulating cellular energetics

Hanahan & Weinberg, 2011

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What is the cause of cancer ?
External factors Internal factors

Chemicals Repair failure

Ultraviolet Error
Biology Carcinogens rearrangement

PERSISTENT GENE ABNORMALITY

In the cell nucleus : > 100 protooncogenes

many tumor suppressor genes
Identified in different chromosomes
(chromosome 1-22, X and Y)
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 Viruses and Cancer
Viruses Associated with Human Cancers
Virus Associated tumors

DNA viruses
Epstein-Barr Burkitts lymphoma
Nasopharyngeal cancer
Hepatitis B Liver cancer
Papilloma virus Benign warts
Cervical cancer
RNA viruses
Human immunodeficiency virus (HIV-1) Kaposis sarcoma
Human T-cell leukemia virus Type I (HTLV-1) Adult T-cell leukemia
HTLV-2
HTLV-5 Hairy cell leukemia
Cutaneous T-cell leukemia
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 Effect of EBV gene product
Product Factor mechanism of induction biological effect
LMP1 MMP-9 induces transcription invasiveness, motility
LMP1 COX-2 induces transcription angiogenesis
LMP-1 VEGF induces transcription angiogenesis
LMP-1 FGF-2 induces transcription angiogenesis
LMP-1 HIF-1a induces transcription angiogenesis
LMP-1 MUC-1 induces transcription metastasis
LMP-1 MMP-1 induces transcription collagenase, metastasis
LMP-1 IL-8 induces transcription angiogenesis
BZLF-1 MMP-9 induces transcription degradation of ECM
EBNA-3C Nm23-H1 interacts with tumor supressor
protein, NM23-H1 metastasis
LMP-2A mTOR increases protein translation
ERK growth related genes & down motility, metastasis
reg. epith diff
Effect of HPV Protein E6 & E7 on
the Cell Cycle

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 Nutritionally Linked Cancers
Carcinogen Promoter
Site Inhibitors
From From Mechanism

Esophagus Pickled, salted ? Yellow-green

foods vegetables
Stomach Pickled, smoked Salt Atrophic Yellow-green
foods, nitrate gastritis vegetables
Liver Mycotoxins, Hepatitis Cytotoxicity ?
nitrosamines antigen
Senecio alkaloids, Alcohol Cytotoxicity
hepatitis antigen
Colon Fried foods Fats Bile acids Bran fiber,
calcium ion,
vegetables
Breast Fried foods Fats Hormonal Same as
imbalances above
Prostate Fried foods? Fats Hormonal ?
imbalances
Endometrium ? Fats Obesity, ?
and ovary estrogen 14
 Irreversibility or Reversibility of Carcinogenesis
as a Function of Action Mechanism

Type of Type and Mode of Action

Reversibility Evidence
Cancer of Carcinogen

Glandular, Genotoxic from salted Poor Migrant studies:

Stomach pickled foods maintenance of
risk in migrants

Lung Small amounts of Good People who stop

genotoxic carcinogens, smoking have
polycyclic aromatic progressively
hydrocarbons and lesser risk
nicotine derived
nitrosamines, large
amounts of phenols,
catechols, terpenes,
epigenetic cytotoxic
enhancing agents
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Chromosomes and Cancer
Primary Chromosomal Aberrations in
Hematological Malignancies
Acute Myeloid Abnormality
Leukemia (AML)
t(1;7)(p11;p11) del/t(12p)
Trisomy 4 t(15;17)(q22;q11)
Monosomy 5 inv(16)(p13;q22)
t(6;11)(q27;q23) del(16)(q22;q24)
Monosomy 7 I(17q)
Trisomy 8 del(20)(q11;q13)
t(9;11)(p21;p23) Trisomy 21
t(10;11)(p14;q13) Trisomy 22
Trisomy 11

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Acute Lymphocytic Abnormality
Leukemia (ALL)
t(1;11)(p32;q23) t(10;14)(q24;q11)
t(1;19)(q23;p13) t(11;19)(q23;p13)
t(4;11)(q21;q23) del(12)(p11;p13)
t(8;14)(q24;q32) t(11;14)(p13;q11)
t(8;22)(q24;q11) t(1;19)(q23;p13)
t(2;8)(p12;q24) Trisomy 21
t(9;22)(q34;q11)

Abnormality

Chronic Myeloid Leukemia (CML) t(9;22)(q34;q11)

Chronic Lymphocytic Leukemia (CLL) Trisomy 12

t(8;14)(q24;q32)
Lymphoma t(8;22)(q24;q11)
t(2;8)(p12;q24)
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 Primary Chromosomal Aberrations
in Solid Tumors
Tumor Abnormality

Lipoma t(3;12)(q27-28;q14-15)
Ewings sarcoma t(11;22)(q24;q12)
Renal carcinoma t or del(3)(p11-21)
Wilms tumor t or del(11)(p13)
Bladder carcinoma Changes of chromosome 1, i(5p)
Breast cancer Changes of chromosome 1, t or del(16q)
Ovarian cancer Changes of chromosome 1
Germ cell tumor of testis i(12p)
Meningioma Monosomy 22, del(22q)
Neuroblastoma del(1)(p13-32)
Retinoblastoma del(13)(q14)
Malignant melanoma t or del(6q)/i(6p)
t or del(1)(p12-22)
Uterine carcinoma Changes of chromosome 1
Note : t, translocation; del, deletion; i, isochromosome 18
Hereditary forms of cancers

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Adenoma-Carcinoma Sequence

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 Control cell proliferation
and differentiation

ONCOGENE

TUMOR SUPPRESSOR GENES

Control growth
arrest and
apoptosis

21
 Genes involved in cancer
Neoplastic transformation involves :
oncogene
tumor suppressor genes
mutator genes
apoptotic
Activation of oncogenes :
play a predominant role in the formation of cancer
development

Inactivation of tumor suppressor genes :

multiple site of loss of heterozygosity are most
prevalent forms of cancer 22
How does cancer occur ?
External factors Internal factors

Protooncogene Oncogene
Tumor Suppressor Gene Inactive

TUMOR

When ???
1. Deletion / Point Mutation
2. Gene Amplification
3. Chromosome Rearrangement

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Protoonkogen Onkogen
PROTOONKOGEN

Five types of proteins encoded by proto-

oncogenes participate in control of cell
growth:
Class I: Growth Factors
Class II: Receptors for Growth Factors
and Hormones
Class III: Intracellular Signal Transducers
Class IV: Nuclear Transcription Factors
Class V: Cell-Cycle Control Proteins
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 Products of Proto-Oncogenes
Growth Factors Growth Factor Receptors

PDGF ERBB1 ERBB2 HER2/NEU FMS ROS

EGF, FGF TRK IGF-1R RET EEK
IGF 1 and 2 BEK MET SEA KIT
TGF and FGFR1-4 FIG EPH ELK
Signal Transducers
Membrane associated/guanine nucleotide binding
RAS GSP GIP
proteins
Membrane associated/cytoplasmic protein tyrosine ABL SRC FES
kinases FGR SYN LCK

Cytoplasmic protein serine-threonine kinases

RAF MOS COT

Nuclear Transcription Factors

MYC FOS JUN MYB SKI EVI1 REL 26

 Pathway of signal transduction

PDGF receptor Ras proteins PDGF [ s/s ]

EGF receptor (erbB) [ H-ras ] Src protein EGF
M-CSF receptor (fms) [ N-ras ] kinase [src] M-CSF
[ K-ras ] Growth factors

Growth-factor
GTP-binding Membrane/cyto-
receptors acting
proteins skeleton-associated
via tyrosine-
specific protein- tyrosine-specific [ myc ]
kinase activity protein kinases [ fos ]
[ jun ]
Thyroid hormone
[ fes ] [ raf ]
receptor [ erbA ]

Nuclear protein

Cytoplasmic Steroid-type Serine/threonine-

tyrosine-specific growth-factor specific protein
protein kinases receptors kinases
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 Sinyal transduction

Summary of 40 years of Hanahan & Weinberg,

research (1971-2011) 2000) 28
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 / inappro-priate production of the
structurally normal growth-promoting protein
- Chromosomal translocations
- Gene amplification
 Point Mutation
ras oncogene the best 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
- 10%: most ovarial and breast tumors
Model for Action of ras gene
 Chromosomal
translocations

Burkitts lymphoma

Chronic Myelogenous
Leukemia
Gene Amplification

Amplification of N-myc gene

in neuroblastoma: double
minutes/HSR (Homogenous-
staining region)
Tumor NF-1

Suppressor
Gene
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 Tumor Suppresor Gen

Negative regulator of cell cycle & involve in growth

arrest
Knudsons Two-Hit hypothesis (1975) the
development of several human cancer is thought
to involveloss of heterozygosity
Tumor Suppressor Genes (LOH) of tumor
suppressor gene
Rb gene, p53, HNPCC, PAC, DCC

genes involved in the control of abnormal cell proliferation

genes loss or inactivation malignancy
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 Tumor Suppressor Genes in Hereditary
Cancers : The Retinoblastoma Model

The prototype for studies on hereditary cancer is :

RETINOBLASTOMA
a childhood cancer
occur in two forms
affects the retina

Studies of the polymorphic enzyme, esterase D, which had

previously been mapped to 13q14, supported the evidence for
RB1 (retinoblastoma gene) in this area.

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38
Hereditary retinoblastoma :

close linkage was demonstrated between esterase D alleles

and retinoblastoma
suggesting that the two genes had to be close

Non-hereditary retinoblastoma :

20% of retinoblastomas but not the constitusional DNA of

the patient have an abnormality
usually absence or a deletion in one copy of chromosome 13
reduced levels of esterase D in the tumor

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Deletion of 13q14 in Retinoblastomas

The phosphorylation
state of the RB
Position of protein varies
RB1 gene throughout the cell
cycle and correlates
with its capacity to
interact with the
transcription factors
and with viral
Chromosome Chromosome 13 oncoproteins.
13 deleted for the
region containing
the RB1 gene 40
 RB
G0 No transcription
Mode of Action E2F DP-1

of RB
RB
G1 E2F
No transcription
DP-1
Cyclin D1/CDK4-6
Transcription
Cyclin E/CDK2 P P
RB E2F DP-1
S
TTTCGCGC
P
Cyclin A/CDK2
P P Phosphorylation
During G1, cyclin D1/CDK4-
of RB is
RB
G2 During
6
maintained
and cyclin
G0 and
by
E/CDK2
Gcyclin
1 RB is
underphosphorylated
phosphorylate
A/CDK2 until mitosis
RB andwhen
and
E2F-
P
is is
1
it bound
released
dephosphorylated
to the
to interact
E2F-1
transcription
with
ready and
either
promote
tofactor
re-enter G1
Dephosphorylation M RB complexed
transcription
or to go intowith
the
fromDP-1.
stationary
genes
necessary for S phase.
phase.

RB
G0 G1 E2F
No transcription
DP-1 41
Tumor suppressor p53
P53 halts progression
when DNA damaged
to give cell time to
repair or
triggers apoptosis of
damaged cell by
activating Bcl-2 causing
mitochondria to
release cytochrome C
and activate caspase
system
If damaged (mutated)
cell moves to S phase
then it may replicate
DNA lesion
Mode of Action of p53
p53 gene

? p53
?
STABILIZATION

WAF1/CIP1 gene

p21 APOPTOSIS

DNA REPAIR
CYC CYC
CDK CDK
STOP
Kinase No kinase G1 S
GO

activity activity
M G2
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 Apoptosis

Program Cell Death (PCD)

Genetically controlled unwanted cell
- during morphogenesis
- during proliferation and differentiation
Differs from necrosis / accidental cell death
- active process
- no surrounding tissue damage or induction of
inflammatory responses
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APOPTOSIS

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Regulation of cell death

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THE OUTLINE OF APOPTOSIS

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 TARGET STRATEGIS TERAPI KANKER

Memicu Apoptosis Sel Hambat Proliferasi

1 Kanker 2 Sel Kanker

Hambat Transduksi
Siklus Sel Sinyal
Pertumbuhan

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ChemoTx research
<1990 1980-2000 2000-2010 >2010
Sifat Eksploratif Eksploratif Targeted Targeted
Strategi Screening Activity Guided Metabolomics AB-conjugate
screening based-
screening
Jangka waktu > 30 th 10-20 th < 10 th < 5 tahun

Efisiensi 1 : >1000 1 : - 1000 1: <100 1: -20

Output NCE NCE NCE/ekstract AB-conjugate

Outcome Broad spectrum Broad spectrum Specific marked Specific marked

tox tox cancer cancer

Contoh Taxol Doxorubicin Brucein/Brucea AB Her2-Taxan

japanica
HER-2 as Target Tx Ca Mammae

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