Molecular Carcinogenesis

Krisna Murti

Department of Anatomical Pathology, Faculty of Medicine,

University of Sriwijaya
Leading causes of death in U.S.

Death Rate

1600
1400
Deaths per 100,000

1200 Total
1000 Heart
Cancer
800
Stroke
600
Accident
400
200
0
1950 1960 1970 1980 1990 2000
Year
from CDC
 Change in causes of death
Rate Per 100,000
600 586.8
1950
500
2000
400

300 258.2

180.5 193.7 200.9

200

100 60.9 48.1

23.7
0
Heart Cerebrovascular Pneumonia/ Cancer
Diseases Diseases Influenza
* Age-adjusted to the 2000 US standard population.
Source: US Mortality Volume 1950, National Vital Statistics Report, 2002, Vol. 50, No. 15.
 Invasive cancer versus age
Invasive Cancer Incidence in U.S.

3500

3000

2500
per 100,000

2000

1500

1000

500

85+
10-14
15-19
20-24
25-29
30-34
35-39
40-44
45-49
50-54
55-59
60-64
65-69
70-74
75-79
80-84
<1
1-4
5-9

Age at Diagnosis (Years)

data from National Cancer Institute

http://www.cdc.gov/cancer/npcr/uscs/report/
Cancers by type and geneder

American Cancer Society

 Cancer death rates
MALE FEMALE

American Cancer Society

Etiologies?
When you shake hands with me!

1775 – Percival Pott discovers “occupational cancer”

of scrotum in chimney sweeps and in hands
of gardeners who spread coal tar
 Coal tar causes skin cancer

1891 -- Katsusabura Yamagiwa shows that coal tar

causes skin cancer when painted on rabbits’ ears.
 Radiation causes cancer

1908 – Clunet shows that X-rays cause cancer in animals

X-rays are mutagens
Human migration and cancer

Rubin and Farber, Pathology

 Same virus, different outcomes

EBV

Nasopharyngeal
Mononucleosis Burkitt’s Lymphoma
Cancer

Immune Suppression
Malaria Dietary Factors
AIDS
Organ Transplants
 Known causes of human cancer

• Chemical Exposure
– Tobacco smoke
– Environmental (Polychlorinated
biphenyls)
– Occupational (coal tar,
asbestos, aniline dye)
– Diet (aflatoxin)
• Radiation (UV, ionizing)
• Infection
– Viruses (EBV, hepatitis B,
papilloma)
– Bacteria (Helicobacter)
• Inherited familial cancer syndromes
 Diagnosis of neoplasia (basic)

Symptoms Screening
Incidental
Weight loss Pap smear
Radiology
Rectal bleeding Mammogram
> ~1 gm (109 cells)
Persistent cough Occult blood

Biopsy

Histopathology

Autopsy

Staging
 Cancer arises from single cells

metastatic adenocarcinoma within lymphatic

vessel in lung (WebPath)

1858 – Rudolf Virchow proposes that “omnis cellula e cellula”

All cells come from cells
Metastatic cancer cells resemble the primary
All cells of a cancer come from a single cell
 Clonality of cancer

• Cancers are usually clonal in origin

– X-inactivation studies in human
cancer

• Transformation can be observed in

cell culture
 Initiators and promoters

• Tumor Initiators = Mutagens

– X rays
– Ultraviolet Light
– DNA alkylating agents

• Tumor Promoters = Proliferation Inducers

– Phorbol Esters (croton oil)
– Inflammation (hepatitis)
– Estrogens and Androgens
– Epstein-Barr Virus
 Genetic theory of cancer

dispermic fertilization in sea urchin

Theodor Boveri, 1914

normal cancer
IF by Bill Brinkley
Which genetic changes?
Molecular basis of cancer
 Cancer is a genetic disease

• Somatic mutations occur in most cancers

• Inherited germline mutations occur in rare
familial cancer syndromes
• Increases in mutation rate or genomic
instability increase frequency of cancer
• Aneuploidy is a hallmark of cancer cells
• Genetic selection at the level of single cells
 Growth fraction
Growth Doubling
Experimental tumors Fraction (%) Time (days)

L1210 (mouse) 86 0.5

B16 (mouse) 55 1.9
LL (mouse) 38 2.9
DMBA (rat) 10 7.4

(hours)
Human tumors
Embryonal carcinoma 90 27
Lymphoma (high grade) 90 29
Squamous cell carcinoma 25 58
Adenocarcinoma 6 83

Normal Human Bone Marrow 35 --

Theories of cancer genesis
Chromosomal changes in the genome of
cancer cells: iceberg tip
Nucleotide changes in the genome of cancer cells:
unseen site of the iceberg

a change in one base

when a base is added to the sequence

when one or more bases in the sequence is

replaced by the same number of bases
when a base is deleted from the sequence
 Tumor clonality by X-inactivation

Heterozygous Female Zygote Monoclonal Tumor

[single G6PD isoenyzme]
X A XB
OR

AB
Random Inactivation
of X Chromosomes
Malignancy
During Early Development AB

Polyclonal Tumor
[two G6PD isoenzymes]
Tumor clonality as a diagnostic

• Immunoglobulin and TCR genes

rearrangement

• Rearrangements are unique in each cell

• Rearrangements display allelic exclusion

Monoclonality of cancer
 Clonality of lymphoid proliferation

Cell Type Benign Malignant

B Lymphocyte Ig Light Chain Ig Kappa or

Heterogeneity Lambda Only

Plasma Cells Heterogeneous Ig Monoclonal Ig

Electrophoresis Spike

T Lymphocyte Heterogeneous Homogeneous

Variable Regions Variable Regions
Multistep process of cancer growth
Responsible genes in cancer
development
Responsible genes in cancer
development
 Oncogenes

• Oncogenes code for oncoproteins

• Mutation of proto-oncogenes

• Proto-oncogenes code for cellular proteins

which regulate normal cell growth and
differentiation
o Growth factors
o Growth factor receptors
o Signal transduction proteins (RAS)
o Nuclear regulatory proteins
o Cell cycle regulators
 Oncogenes

• Proto-oncogenes = ras

• Oncogenes = mutated ras

• Always activated

• Always stimulating proliferation

Important oncogenes
Important tumor–suppressor anti-
oncogenes
Molecular basis of cancer
Protein encoded by proto-oncogenes
participate in control of cell growth
Functions of cellular proto-oncogenes
Functions of cellular proto-oncogenes

Amino acid substitutions in Ras family proteins (inactivates GTPase)

Proto-oncogenes → oncogenes
Oncogenes are usually dominant
(gain of function)
Oncogenes are usually dominant
(gain of function)
Gene amplification
Chromosomal rearrangements or
translocations
Tumor suppressor genes
Tumor suppressor genes
Rb genes
p53
Cell cycles
Two of the most important
internal factors are kinases
and cyclins
Kinases and Cyclins
DNA repair genes
 Molecular
mechanisms of
DNA double
strand break
repair
Important of DNA repair
Translocation and Bcr-Abl fusion in
CML
Activation of proto-oncogenes
Multistep carcinogenesis
Cancer-associated genes

Red: Proto-oncogenes
Blue: Cancer suppressor genes
Green: DNA repair genes
Purple: Genes regulates apoptosis
Stem cells as the target of carcinogens
Activation mechanisms of proto-
oncogenes
Activation mechanisms of proto-
oncogenes
Thousands of targets
Kinase growth factor pathway
Activated receptor

Tyrosine kinase receptor Ligand

binding Cell membrane

Tyrosine
kinase Signal
domain
transduction

Proliferation Survival Migration

Tumour growth
and metastases Original by Dr. Axel Ullrich
EGFR signaling

• The EGFR is activated

by growth factors (e.g. epidermal
growth factor (EGF) and
transforming growth factor-
(TGF-)

• EGFR-activation leads to
the building of either receptor
homo- or hetero-dimers

• Receptor dimerization initiates

# Intracellular signaling cascade
# Gene activation
# Stimulation of cell cycle
progression
 EGFR signaling

* Baselga 2002
EGFR2/HER2 signaling
Mechanisms of HER2 overexpression
Uncontrolled growth of cancer
Tumor progression
Metastasis mechanism
Pathological diagnosis of cancer

• Histology
• Cytology
• Histochemistry and cytochemistry
• Immunohistochemistry
• Electrone microscopy
• Tumor markers
• Flow cytometry
• Molecular diagnostic techniques
• DNA microarray analysis of tumor
Malignant versus benign tumors
Invasion and metastasis
 Hallmark of cancer

Hanahan and Weinberg, 2000

 Hallmark of cancer

Hanahan and Weinberg, 2011

 Signaling interactions in the tumor
microenvironment during malignant progression
Molecular mechanisms:Targeted therapy
development
Resume
Thank you

Anger may be frozen

But the sun of forgiving
Can make it melted

By Gede prama