Molecular Perspectives

In Carcinogenesis

Dolores V. Viliran, MD

Department of Biochemistry & Nutrition

FEU-NRMF,INSTITUTE OF MEDICINE
INTRODUCTION

CANCER
Cancer is an overgrowth of cells bearing
cumulative genetic injuries that confer growth
advantage over the normal cells [Nowell’s
Law]

Cancer cells can be characterized as

antisocial, fairly autonomous units that
appear to be indifferent to the constraints
and regulatory signals imposed on
normal cells [Robbin’s]
 CANCER CELLS AND NORMAL
CELLS
CANCER CELLS NORMAL CELLS
Frequent
mitoses

Normal
cell

Nucleus

Few
Blood vessel mitoses

Abnormal
heterogeneous cells

Loss of contact inhibition Oncogene expression is rare

Increase in growth factor secretion
Increase in oncogene expression
Loss of tumor suppressor genes
Intermittent or co-ordinated
growth factor secretion
Presence of tumor suppressor
genes
 CHARACTERISTICS OF
CANCER
• Clonality

• Autonomy

• Anaplasia

• Metastasis
 CHARACTERISTICS OF CANCER
Clonality
• Cancer is a genetic disease at the cellular level.
• Genetic mutations play a critical role in
pathogenesis of cancer.
• Consequences of genetic instability:
– Phenotypic heterogeneity
– Tumor progression
• Proto-oncogenes and oncogenes
• Dominant mutations = mutation resulting from
conversion of protooncogenes to oncogenes
• Recessive mutations = mutation resulting from
damage or loss of tumor suppressor gene.
 Cancer Genetics
• Tumors arise as clones from a single
cell. At the cellular level, cancer is a
genetic disease.
• The development of the malignant clone
is due to mutations in DNA due to:
– Random replication errors
– Exposure to carcinogens
– Faulty DNA repair process
 Evidence that Mutations Cause
Cancer
• Recurring sites of chromosome change are
observed in cancers at sites of genes
involved in cellular growth control.
• Most carcinogens are mutagens.
• Defects in DNA repair systems increase
the possibility of cancer.
 CHARACTERISTICS OF CANCER
Autonomy
• Cancer cells are able to proliferate despite
regulatory influences.
• Unrestricted proliferation results in tumor
formation.
• Mechanisms:
– Growth factor secretion
– Increased number of cell receptors
– Independent activation of key biochemical
process
• Proliferation depends on the cell cycle.
 AUTONOMY
• Brought about by mutations in the cell’s
genetic apparatus
• Most common in tissues with rapid
turnover, especially:
- those exposed to environmental agents
- those whose proliferation is hormone-
dependent
• Proliferation is dependent on the cell
regeneration cycle
 The Cell Cycle
DEATH
G0
DIFFERENTIATION

G2/M checkpoint

Mitosis
M
DNA content = 2n

DNA content = 4n
G2 G1

S G1/S checkpoint
DNA synthesis
CYCLIN, CDK,CDKI:
PHOSPHORYLATION
 Cell Cycle Regulation
• Process assures that cell accurately
duplicates its contents.
• Important checkpoints are present at
G1 and G2 and are regulated by
protein kinases called cyclins (cdk).
• Checkpoints determine whether the
cell proceeds to next phase of the
cycle.
 G2/M Checkpoint
• Regulated by the cyclin B/cdc2
(mitosis promoting factor or MPF).
• Activity of this cyclin with its
substrate results in:
– Chromosome condensation
– Nuclear membrane breakdown
– Spindle formation
 G1/S Checkpoint
• Area most often disrupted in cancer.
• Mechanism of regulation is complex
and involves the phosphorylation of
the Rb gene. This results in:
– Activation of several genes needed for S
phase progression.
– Promotes differentiation through
association with transcription factors.
Rb Gene Activation
 Cyclin Regulators
• Regulated by cdk inhibitors (cdki).
• May be induced by growth inhibitors
and inhibited by positive growth
factors.
• Genetic alterations in cdki occur with
high frequency in some cancers.
 Cyclin Regulators
• p 21: inhibits cell cycle progression and
permits DNA repair to take place.
• P53: “the guardian of the genome”
– In the presence of DNA damage, influences
transcription to either:
• Halt cell cycle progression to facilitate DNA repair.
• In cases of severe DNA damage, activates apoptosis.
– Mutations in p53 are the most common genetic
alterations found in human cancer.
 CHARACTERISTICS OF
CANCER: Anaplasia
• Loss of differentiated function
resulting to bizarre-looking cells
• Large nuclei, prominent nucleoli,
increased chromatin
• Increased and/or abnormal mitosis
• Aneuploidy
• Partial or complete loss of normal
architecture
 Invasion and Metastasis
• The defining characteristic of a
malignancy.
• Invasion: active translocation of
neoplastic cells across tissue
barriers.
• Critical pathologic point: local invasion
and neovascularization. These events
may occur before clinical detection.
ATTRIBUTES OF CANCER

Metastasis
Two basic steps:
Destruction of the BM
Attachment to the laminin of distant BM

Genes up-regulated among good metastasizers:

EDGF receptor
Basic Fibroblast Growth Factor
Type IV Collagenase
ε-Cathepsin (under-expressed)
Cathepsin B (a lamininase)
Heparanase
 Angiogenesis
• Process of new blood vessel formation.
• Clinical importance:
– Tumor vessel number correlates positively with
risk and degree of dissemination.
– Several cytokines that stimulate endothelial
cell proliferation also stimulate proliferation of
malignant cells.
INVASION AND METASTASIS
 Triad of Invasion
• Adhesion with the basement
membrane

• Local proteolysis

• Mobility and ability to translocate

through dents in body’s structural
barriers
MOLECULAR CARCINOGENESIS

Mutation

the molecular hallmark of most forms of cancer

Gene Families in Cancer Development

1 - Oncogenes
2 - Tumor Suppressor genes
3 - Mutator genes
 Cancer Genes
• Proto-oncogenes – normally promote normal
cell growth; mutations convert them to
oncogenes.
• Tumor suppressor genes – normally restrain
cell growth; loss of function results in
unregulated growth.
• Mutator or DNA repair genes – when
faulty, result in an accumulated rate of
mutations.
ONCOGENE FAMILY

+ oncogenes

Oncogenes
promote cell proliferation
dominant & highly conserved
types: viral oncogenes [v-oncs]
cellular oncogenes [c-oncs]
Proto-oncogene ⇒ “Mutation” ⇒ Oncogene
ONCOGENE FAMILY

Classification of Oncogenes

A. Secreted Growth Factors

c-sis, hst
B. Cell Surface Receptors
erb B, fms, ret, trk, fes, fms Components of
signal transduction
C. Intracellular Transducers pathways
c-src, c-abl, mst, ras
D. DNA-binding Nuclear Proteins
myc, jun, fos

E. Regulators of the Cell Cycle

bcl, bax, bad
SIGNAL TRANSDUCTION
ONCOGENE FAMILY

Mechanisms of Oncogene Activation

1. Point Mutation GTP

H-ras [codon 12] H-ras

Normal CGC → Gly

Bladder ca CTC → Val
Perpetual cell division

2. Gene Amplification
Double minutes
HSRs
Homogenously
Staining regions Normal copy Multiple copies
ONCOGENE FAMILY

Mechanisms of Oncogene Activation

3. Gene Translocation

Ex. Burkitt’s Lymphoma

ONCOGENE FAMILY

Mechanisms of Oncogene Activation

3. Gene Translocation

Ex. Chronic Myelogenous Leukemia [CML]

ONCOGENE FAMILY

Mechanisms of Oncogene Activation

4. Viral Gene Integration

promoter

Viral promoter
TUMOR SUPPRESSOR GENE
FAMILY
TS Genes
inhibit growth and multiplication of mutated cells
prevent neoplastic transformation
recessive & highly conserved

Classification of TS genes
A. Cell Adhesion Molecules
APC, DCC

B. Regulators of the Cell Cycle

RB1, Tp53
TUMOR SUPPRESSOR GENE
FAMILY
KNUDSON’S Two-Hit Hypothesis

1st Hit: TS mutation or Inherited mutation

2nd Hit: gross chromosomal loss
TUMOR SUPPRESSOR GENE
FAMILY
Retinoblastoma gene [RB1 gene]
rare form of childhood malignancy
forms: hereditary & sporadic

pRb

105-KDa nuclear protein

inhibits E2F [prevents G1 → S transition]
inhibited by: phosphorylation
viral oncoproteins [E1A, HPV E7]
TUMOR SUPPRESSOR GENE
FAMILY
Tp53 gene

location: 17p13.1
product: p53 protein [53 KDa]
function: induces DNA repair or apoptosis
mutation: point mutation > deletion
results to: loss of function & extended lifespan of p53
Clinical conditions: carcinomas, Li Fraumeni
Syndrome
p53 inhibited by: E1B, HPV E6, mdm2
TUMOR SUPPRESSOR GENE
FAMILY
p53 protein
p53 in action
MUTATOR GENE FAMILY

Mutator Genes
involved in ensuring the fidelity of replication
function: checks for & corrects mismatched pairs
mutation ⇒ inefficient repair & replication leading
increased propensity of oncogenes and tumor
suppressor genes to undergo mutation
first described in E coli [Mut-HSL system]
Fischel, et al = Human homologs
leads to the formation of Microsatellite Instability
[MIN+]
In summary …..

MUTATOR GENES

ONCOGENES TS GENES
Re-cap of Molecular
Carcinogenesis

Proto-oncogene Gain-of-function
TS gene Loss-of-function CANCER

Mutator gene Loss-of-function

 CARCINOGENS
• Occupation related causes
• Lifestyle related causes
– Tobacco
– Diet
– Sexual practices
• Multifactorial causes
• Viral carcinogens
• Chemical carcinogens
• Ionizing radiation
• X-rays
• Stress Sources of Free
• Toxins
• Sunlight Radicals
• Solvents
• Pollution
• Cigarette
• Pesticides Smoking
10 Quad Trillion free radicals per
• Herbicides cigarette!
• Medications
• Airline travel
• Radioactivity
• Food additives R.I.P
• Polluted Foods
• High heat cooking They only said it
was dangerous.
• Synthetic materials
They didn’t say
• Household cleaners
it could be
• Environmental Chemicals
lethal.
Lots more…
MENU
 Occupational Risk Factors
Etiology Site of Malignancy
Arsenic Lung, skin, liver
Asbestos Mesothelium, lung
Benzene Leukemia
Benzedine Bladder
Chromium cpds Lung
Radiation (mining) Numerous locations
Mustard gas Lung
Polycyclic hydrocarbons Lung, skin
Vinyl Chloride Angiosarcoma of liver
 Lifestyle Risk Factors
Tobacco-related:
• Lung cancer
• Pancreatic cancer
• Bladder cancer
• Renal cancer
• Cervical cancer
 Diet-Related Risk Factors
Nitrates
Salt
Low vitamins A, C, E Gastric Cancer
Low consumption of Esophageal
yellow-green Cancer
vegetables
 Diet-Related Risk Factors
High fat Colon Cancer
Low fiber Pancreatic Cancer
Prostate Cancer
Low calcium
Breast Cancer
High fried
Uterine Cancer
foods

Mycotoxins Liver Cancer

 Sexual Practices Risk Factors

Sexual promiscuity
Multiple partners
Unsafe Sex Cervical Cancer
Human Papillomavirus
 Multifactorial Factors
Oral Cavity Cancer
Tobacco + Alcohol
Esophageal Cancer

Tobacco + Asbestos
Tobacco + mining Respiratory
Tract Cancer
Tobacco + uranium + Lung Cancer
radium
CARCINOGEN METABOLISM

Three Main Categories:

I. Chemical Carcinogens
II. Physical Carcinogens
III. Viral Agents

Carcinogens Mutations Cancer

Environmental ?
factors
CHEMICAL CARCINOGENESIS

Stages:
Initiation - primary exposure

Promotion - transformation

Progression - Cancer growth

Frank Cancer
CHEMICAL CARCINOGENESIS

Initiation
normal cells are exposed to a carcinogen
not enough to cause malignant transformation
requires one round of cell division
normal cells are exposed to a carcinogen
1. Direct-acting carcinogens
2. Indirect-acting carcinogens

Cytochrome Ultimate
procarcinogen
P450 carcinogen
CHEMICAL CARCINOGENESIS

Promotion
initiated cells are exposed to promoters
promoters are not carcinogens !
properties of promoters reversible
dose-dependent
time-dependent
Types of Carcinogens …...

1. Direct carcinogens
2. Procarcinogens ⇒ Ultimate carcinogens
CHEMICAL CARCINOGENESIS

Direct-acting Procarcinogens
Carcinogens
PAHs
 cyclophosphamide Aromatic amines & Azo dyes
 chlorambucil Aflatoxin B1
 busulfan Nitrosamine & Amides
 melphalan Asbestos
Vinyl chloride
Promoters
 saccharine & cyclamates Chromium, nickel, other metals
 Estrogen Arsenic
 Diesthystilbestrol [DES]
 Physical Carcinogenesis
• Radiation-induced mutation in the
host cell
• Transmits irreversible changes in
gene expression to cell progeny
 Sources of Potentially
Carcinogenic Radiation
• Sunlight
• Artificial sources of UV light
• X-rays
• Radio-chemicals
• Nuclear fission
PHYSICAL CARCINOGENESIS

Ultraviolet Rays

UV-A = 320 - 400 nm

UV-B = 280 - 320 nm
UV-C = 200 - 280 nm
PHYSICAL CARCINOGENESIS

Ultraviolet Rays
UV-C ⇒ filtered by ozone

UV-B

Inhibition of cell division

inactivation of enzymes
induction of mutations
cell death at high doses

Squamous cell cancer

Basal cell cancer
Melanocarcinoma
PHYSICAL CARCINOGENESIS

Ionizing Radiation
includes electromagnetic rays & particulate matter
mechanism: ↑ free radicals & mutations
pathology: leukemias > thyroid ca > lung & breast ca
resistant tissues: bone, skin and the GIT

PRE-IRRADIATION POST-IRRADIATION
 Viral Carcinogenesis
• Viral carcinogens are classified into
RNA and DNA viruses.

• Most RNA oncogenic viruses belong to

the family of retroviruses that
contain reverse transcriptase
mediates transfer of viral RNA into
virus specific DNA.
 Viral Oncogenes
RETROVIRUS

Oncogene
Oncogene Viral RNA Oncogene
REVERSE TRANSCRIPTASE protein
Viral DNA

RNA messenger TRANSCRIPTION

NUCLEUS INSERTION

DNA

TRANSCRIPTION Viral genome

Viral RNA Oncogene

CELL
MEMBRANE CYTOPLASM
 Viruses Associated With The
Development Of Human Neoplasia
VIRUSES NEOPLASMS

DNA VIRUSES
Human papilloma virus Cervical Ca, warts, ano-
genital carcinoma
Herpes simplex virus II Cervical carcinoma
Epstein-Barr virus NPCa, African Burkitt’s
Herpes simplex virus 8 Kaposi’s sarcoma
Hepatitis B virus Hepatocellular Ca
Herpes simplex virus 6 Certain B cell
(HBLV) lymphomas
 Viruses Associated With The
Development Of Human Neoplasia

VIRUSES NEOPLASMS

RNA VIRUSES
Human T-cell leukemia virus I Some T-cell leukemia,

lymphoma
Human T-cell leukemia virus II Some cases of hairy

cell leukemia
Human immunodeficiency virus I Lymphoma; Kaposi’s
sarcoma
VIRAL AGENTS: DNA viruses

Human Papillomavirus [HPV types 16, 18, 31, 33 & 35]

Interruption of the E1/E2 ORF

E2 is not expressed

Over-expression of E6 & E7
VIRAL AGENTS: DNA viruses

Epstein-Barr Virus [EBV]

in Burkitt’s, B-cell & Hodgkin’s lymphomas + NP ca
tropism: CD21+ cells [e.g., B cells, epithelial cells]
mechanism: viral entry ⇒ episomal existence ⇒ latency
⇒ (+) LMP-1, EBNA-1, EBNA-2 ⇒ immortalization

Hepatitis B virus [HBV]

induction of chronic hepatocyte injury ⇒ (+) HBx

HBx activates protein kinase c for transformation
VIRAL AGENTS: RNA viruses

Human T-cell Leukemia Virus [HTLV]

a retrovirus
tropism: CD4+ cells
mechanism: Tax protein

↑ transcription ↑ c-fos, c-sis, IL-1 and IL-2

Viral replication T cell proliferation

 Principal Pathways of
Malignancy
1. Proliferation
2. Cell-Cycle Progression
3. DNA Repair
4. Immortalization
5. Apoptosis
6. Angiogenesis
7. Metastasis and Invasion
SIGNAL TRANSDUCTION
 PROLIFERATION
(Growth Factor Signaling Pathway)
• Uncontrolled and uncoordinated
proliferation
• Uncontrolled growth stimulated by:
3. Increased secretion of Growth Factors
(PDGF,EGF,FGF,VEGF,NGF)
4. Increased Growth Factor receptors
5. Independent activation of certain enzyme
and protein production pathways
 PROLIFERATION
(Growth Factor Signaling Pathway)
• Receptor Tyrosine kinase Pathway
(RTK)-Main pathway
• RTK ligands: NGF PDGF FGF EGF
• Functions of RTK:
4. promotion of cell survival
5. regulation of cell proliferation and
differentiation
6. modulation of cellular metabolism
 PROLIFERATION
(Growth Factor Signaling Pathway)

RTK SIGNALING PATHWAYS

• Ras-MAP Kinase Pathway- most
prominent
• PI3 kinase Pathway
• Phospholipase C Pathway
PROLIF ERATI ON
 PROLIFERATION
(Growth Factor Signaling Pathway)
Therapeutic implications
Blocking of GF mitogenic signaling is achieved
by:
• Preventing binding of GF to receptor or
receptor dimerization with specific agent
• Preventing receptor activation with small
molecule inhibitors
• Blocking cytoplasmic proteins downstream
of the activated receptor pathway
 The Cell Cycle
DEATH
G0
DIFFERENTIATION

G2/M checkpoint

Mitosis
M
DNA content = 2n

DNA content = 4n
G2 G1

S G1/S checkpoint
DNA synthesis
 Cell Cycle Regulation
• Process assures that cell accurately
duplicates its contents.
• Important checkpoints are present at G1
and G2 and are regulated by proteins
Cyclins and Cyclin-dependent Kinases
(CDKs).
• Checkpoints determine whether the cell
proceeds to next phase of the cycle.
 Cyclins and Cyclin-dependent
Kinases (CDKs)
• CYCLINS – activate protein kinases
• CDKs – protein enzymes which
selectively phosphorylate specific
serine/threonine residues in their
substrates
• Dimeric complex with
catalytic subunit (CDK 1-9)
regulatory subunit (Cyclin A-H,T)
 G2/M Checkpoint
• Regulated by the cyclin B/cdc2 (mitosis promoting
factor or MPF).
• Regulated mainly by intracellular signal
(Completion of DNA Synthesis)
• MPF is activated by dephosphorylation by cdc25
• Cyclin B is degraded by Anaphase Promoting
Complex (APC)
• Role of G2/M checkpoint: to prevent mitosis when
DNA is damaged and not yet repaired
CYCLIN, CDK,CDKI:
PHOSPHORYLATION
 G1/S Checkpoint
• Area most often disrupted in cancer.
• Mechanism of regulation is complex and
involves the phosphorylation of the Rb
gene.
• Regulated by extracellular signals (e.g. GF)
• “R” point (restriction)- point late in G1
beyond which cell cycle progression
becomes independent from external GF
• Regulated mainly by CDK4/cyclin D
Rb Gene Activation
 Cyclin Regulators- CDK
Inhibitors
• CDK inhibitors – inhibit the activity of CDK-cyclin
complex
• Two Groups:
1) INK4 family – p15 16 18 19
2) CIP-KIP family – p21 p27
Actions:
P15- change response to anti-mitogenic agents
P16- inhibits CDK4/cyclin D
P19- induces p53 stabilization
P21-induces cell cycle arrest via activation by p53
P27- inhibits CDK2/cyclin E
 Cyclin Regulators
• p 21: activated by p53 inhibiting cell cycle
progression and permitting DNA repair to
take place.
• P53: “the guardian of the genome”
– In the presence of DNA damage, influences
transcription to either:
• Halt cell cycle progression to facilitate DNA repair.
• In cases of severe DNA damage, activates apoptosis.
– Mutations in p53 are the most common genetic
alterations found in human cancer.
p53 in action
 CELL-CYC LE
PR OG RES SION
 Clinical Significance
Oncogenic alterations in cell cycle regulators:
• Loss of p53 and pRB function as tumor
suppressors
• Increased expression of Cyclin D1(Mantle Cell
Lymphoma)
• CDK4 amplification in sarcomas, glioma
• Mutations in p16-binding domain of CDK4(Familial
Melanoma)
• Inactivation of INK4
• Alterations in Cyclin D1,p16
• Decreased levels of p27 (Breast Ca)
• Over expression of cdc25
 Therapeutic Implications
Approaches using Inhibitors of CDKs
as therapeutic agents
• Small molecules
• Protein therapy
• Antisense
• Gene therapy
Most cytotoxic agents block the cell
cycle in the S/G2/M phases
 DNA REPAIR PATHWAYS
• Cancer as “Malady of Genes”
• Defects in the maintenance of genome
stability
• Repair Mechanisms:
4. Mismatch excision repair
5. Base excision repair
6. Nucleotide excision repair
7. Double strand base repair
 DNA REPAIR PATHWAYS
Clinical Significance
HNPCC – mutations in genes involved in DNA
repair pathways (MSH1 MSH2)
• Somatic defects in repeated DNA elements
leading to Microsatellite instability (MSI)
• Inactivation of TGF-β (tumor suppressor)
• Inactivation of BAX gene
IMM OR TALI ZATION
 Telomeres and Telomerase
Telomeres- specialized structures at
chromosome ends generated and
maintained by telomerase
Telomerase- ribonucleoprotein enzyme which
preserves the integrity of telomeres
* key component in immortalization of
cancer cells
Telomere length- represents a molecular
clock that determines the life span of the
cell
 Telomeres and Telomerase
Clinical Significance
• Most normal adult tissues have NO telomerase
activity
• Telomerase activity is present in 90% of tumors
Therapeutic Implication
hTERT- protein identified to be catalytic subunit of
telomerase
• limiting component of telomerase activity
• can be a target for small molecule inhibitor
 APOPTOSIS
• APOPTOSIS – programmed cell death
• Important in:
3. Steady-state kinetics of normal tissues
4. Focal deletion of cells during normal
embryonic development
5. Seen after chemotherapy and radiation
* Balance between proliferation and apoptosis
is critical in determining growth or
regression
 Components of Apoptotic
Pathway
1) CASPASES (Cysteine-containing aspartate-
specific proteases)
• Initiator Caspases – activated in response to cell
death signal
• Executioner or Effector Caspases- progress the
death signal activating cascade resulting to DNA
fragmentation and cell death
Caspase prodomains – DED CARD
Death ligands – TNF-α , Fas , TRAIL
Survival Signals – NFκβ
 Components of Apoptotic
Pathway
2) CYTOCHROME C – component of mitochondria
released in response to apoptotic signals
3) BCL-2 Family of Proteins- located upstream in the
pathway
• Provides pivotal decisional checkpoint in the fate
of the cell after a death stimulus
• Contains BH1-BH4 domains necessary for
interaction
• Anti-apoptotic – BCL-2 BCL-xL
• Pro-apoptotic – BAX BAD BAK BID
 APOPTOTIC PATHWAYS
1) FAS-mediated apoptosis
• FAS – cell surface receptor of TNF family
which binds to FAS-L
• Eliminates unwanted activated T cells
• Pathway for cytotoxic-mediated signaling
2) P53-mediated apoptosis
• important after chemotherapy and radiation
• Induction of BAX and downregulation of BCL-2
• Induced expression of FAS and DR5
 Clinical Significance
• Over expression of BCL-2 as a prognostic
indicator
• Mutations of BAX in GI Ca and leukemias
• P53 provides a link between cell
proliferation and apoptosis
• Cell survival signals: NFκβ BCL-2
• P53 mutations confer chemoresistance
EVA DING A POPTOS IS
 Therapeutic Implications
• Antisense oligonucleotide against
BCL-2 in the treatment of lymphoma
• BCL-2 antisense as chemosensitizing
agent in solid tumors
• TRAIL ( TNF-related apoptosis
inducing ligand) to induce apoptosis
 ANGIOGENESIS
• Formation of new blood vessels from
existing vascular bed
• Carried out by endothelial cells (EC) and
extra cellular matrix (ECM)
• Regulated by angiogenic factors (inducers
and inhibitors)
* A tumor is unable to grow larger than 1
mm3 w/o developing a new blood supply
Components of Angiogenesis
1) ENDOTHELIAL CELLS
• Fenestrated
• Increased cell adhesion molecules
( E-selectin)
• Increased integrins αγβ3 essential
for viability during growth
• Activated ECs release: bFGF PDGF
IGF-1
Components of Angiogenesis
2) INDUCERS OF ANGIOGENESIS
• VEGF – main inducer
• TGF- β
• TNF-α low concentration - inducer
high concentration -
inhibitor
• PDGF/thymidine phosphorylase
• TGF-α
• EGF
• IL-8
 Components of Angiogenesis
3) CELL ADHESION MOLECULES (CAM)
• Mediate cell-cell adhesion processes
• Selectins
• IG Supergene family- ICAM VCAM
• Cadherins
• Integrins- vitronectin receptor
4) PROTEASES
• Degrade ECM to provide suitable
environment for EC migration thru adjacent
stroma Ex: Metalloproteinases (MMP)
Components of Angiogenesis
1) ANGIOGENESIS INHIBITORS
• Interferon
• TSP-1
• Angiostatin
• Endostatin
• Vasostatin
CLINICAL SIGNIFICANCE:
Tumor angiogenesis switch is triggered as
a result of shift in the balance of
stimulators to inhibitors
ANG IOG ENES IS
 Therapeutic Implications
• Metalloproteinase inhibitors (MMPI) –
block the degradation of basement
membrane
• Inhibitors of endothelial function-
thalidomide, TNP 470,endostatin
• Anti-angiogenic factors – tyrosine
kinase inhibitors of VEGF bFGF PDGF
• Interferon – angiogenic inhibitor
• COX-2 inhibitor – thromboxane A2 as
critical intermediary of angiogenesis
INVASION AND METASTASIS
 Invasion and Metastasis
• The defining characteristic of a
malignancy.
• Invasion: active translocation of
neoplastic cells across tissue
barriers.
• Critical pathologic point: local invasion
and neovascularization. These events
may occur before clinical detection.
PROCESS OF METASTASIS
 Triad of Invasion
• Adhesion with the basement
membrane.
• Local proteolysis
• Mobility and ability to translocate
through rents in body’s structural
barriers.
 ADHESION
• De-regulated function of CAM (E-cadherin)
• Changes in catenin expression leads to loss of
cadherin function
• Integrin over expression in naturally
occurring cancers
• Downregulation of integrin in more advanced
stages of cancer
• Upregulation of ICAM-1 which enhances
extravasation
• Adhesion molecules on EC: E-selectin,VCAM
ICAM
 LOCAL PROTEOLYSIS
• Degradation of basement membrane
to traverse barriers
• Carried out by:
• Serine proteases -uPA elastase
plasmin cathepsin G
• Cysteine proteases- cathepsin B L
• Aspartate proteases – cathepsin D
• Matrix metalloproteinases-
gelatinases interstitial collagenases
stromelysins matrilysins
 MOTILITY
• Tumor cells can move randomly or
directionally toward attractants
• Modulators of motility
GF, hyaluronases, components of ECM,
tumor-secreted factors, host-derived
factors
THERAPEUTIC IMPLICATIONS:
MMPI and monoclonal antibodies
against integrin
METAS TAS IS AND
INV AS ION
Which of the following is TRUE
of carcinogenesis?
A. Carcinogenesis occurs as a result of
genetic mutation secondary to physical and
chemical agents only
B. The ultimate carcinogens are usually
electrophiles which can readily attack NA
C. The most common base involved in
mutagenesis is adenine
D. Tumor suppressor gene is transformed to
oncogene
Which of the following is TRUE
of carcinogenesis?
• Carcinogenesis occurs as a result of
genetic mutation secondary to physical and
chemical agents only
• The ultimate carcinogens are usually
electrophiles which can readily attack
NA
• The most common base involved in
mutagenesis is adenine
• Tumor suppressor gene is transformed to
Tumor p53 suppressor protein is
reffered to as guardian of the
genome bec. it:
A. Enhances the survival of tissues
B. Allows apoptosis to occur on seriously
damaged cells
C. Plays a key role in G2 checkpoint control
D. Arrests the cell cycle at Go phase
Tumor p53 suppressor protein is
reffered to as guardian of the
genome bec. it:
• Enhances the survival of tissues
• Allows apoptosis to occur on seriously
damaged cells
• Plays a key role in G2 checkpoint control
• Arrests the cell cycle at Go phase
 TRUE statements about
oncogenes, EXCEPT:
A. They positively affect cell proliferation
B. Single mutant allele is enough to cause
phenotypic
C. They are mutant protooncegenes
D. Mutation involves a loss in function
 TRUE statements about
oncogenes, EXCEPT:
• They positively affect cell proliferation
• Single mutant allele is enough to cause
phenotypic
• They are mutant protooncegenes
• Mutation involves a loss in function
 This is not a characteristics of
cancer
A. Loss of contact inhibition
B. Uncontrolled proliferation
C. Gain in function of mutator gene
D. Loss of differentiated function
 This is not a characteristics of
cancer
• Loss of contact inhibition
• Uncontrolled proliferation
• Gain in function of mutator gene
• Loss of differentiated function
 TRUE statements about RAS
oncogene activation except:
A. It involves a point mutation in codon 12
B. The mutated RAS results to increased
GTPase activity
C. The mutated gene codes for valine instead
of glycine
D. It is over-expressed in bladder cancer
 TRUE statements about RAS
oncogene activation except:
• It involves a point mutation in codon 12
• The mutated RAS results to increased
GTPase activity
• The mutated gene codes for valine instead
of glycine
• It is over-expressed in bladder cancer
 A biochemical change found in
fast growing tumor cells:
A. Increased catabolism of nucleobases and
nucleotides
B. Inappropriate synthesis of certain growth
factors and hormones
C. An adult pattern of isozymes
D. Markedly decreased glycolysis
 A biochemical change found in
fast growing tumor cells:
• Increased catabolism of nucleobases and
nucleotides
• Inappropriate synthesis of certain
growth factors and hormones
• An adult pattern of isozymes
• Markedly decreased glycolysis
Any Questions ?