Hallmarks of Cancer

Reference:
1. Douglas Hanahan and Robert A. Weinberg, Cell 144, March 4, 2011
2. Douglas Hanahan, Hallmarks of Cancer: New Dimensions. Cancer Discov. 2022
Jan;12(1):31-46
What are ‘ hallmarks of cancer ‘ ?

 Biologic capabilities acquired by cancer cells during the multistep

process of development of human tumors

 Essential Alterations In Cell Physiology That Collectively Lead To

Malignant Growth Of A Normal Cells.
Originally six hallmarks of cancer proposed in 2011:

• Self-sufficiency in growth signals

• Insensitivity to growth-inhibitory (antigrowth)
signals
• Evading apoptosis
• Limitless replicative potential
• Sustained angiogenesis
• Tissue invasion and metastasis
• With development in genetics and epigenetics Hanahan and Weinberg
again redefined “Hallmarks of cancer” in 2011

Two additional hallmarks of cancer are:

• Evading immune destruction.
• Deregulating cellular metabolism
or energetics
Redefined “Hallmarks of cancer” in 2011
2011
Redefined Hallmarks of Cancer: 2022
Hallmarks of Cancer
1. Sustained Proliferative Signalling
• Cancer cells : ‘master of their own destinies’

• Normal cells require growth signals to enter from a

quiescent state into an active proliferative state.

• These signals are transmitted into the cell through

transmembrane receptors that binds to a particular
class of signaling molecules.
• Cancer cells, by deregulating these
signals, become masters of their own
destinies.

• Tumor cells generate their own growth

signals and thereby reducing their
dependence on external stimulation
from their normal tissue
microenvironment.
“The precise identities and sources of the proliferative signals
operating within normal tissues were poorly understood a
decade ago and in general remain so”

Hence, cancer cells to continuously stimulate their own growth

and division signals, leading to uncontrolled cell proliferation
Alternative ways to sustain proliferative signalling

 They may produce growth factor ligands themselves, to which they

can respond via the expression of cognate receptors, resulting in
autocrine proliferative stimulation.

 Alternatively, cancer cells may send signals to stimulate normal cells

within the supporting tumor-associated stroma, which reciprocate by
supplying the cancer cells with various growth factors
1.1 Somatic Mutations in signalling

• Somatic Mutations activate additional downstream pathways that

promote sustained growth

• High-throughput DNA sequencing analyses of cancer cell genomes –

Somatic mutations that constitutive activation of signaling circuits

• RAS-RAF-MAPK PATHWAY
For example:

 40% of human melanomas contain

activating mutations affecting the
structure of the B-Raf protein,
resulting in constitutive signaling
through the Raf to
mitogenactivated protein (MAP)-
kinase pathway
For example:

Mutations in the catalytic subunit of

phosphoinositide 3-kinase (PI3-kinase)
isoforms are being detected in an array
of tumor types, which serve to
hyperactivate the PI3-kinase signaling
circuitry, including its key Akt/PKB
signal transducer
1.2 Disruptions of Negative-Feedback Mechanisms that
Attenuate Proliferative Signaling:

• The importance of negative feedback

loops that normally operate to
dampen various types of signaling and
thereby ensure homeostatic
regulation of the flux of signals
coursing through the intracellular
circuitry
 Defect in negative feed back mechanism leads to uncontrolled
proliferative signaling.

 The prototype of this type of regulation involves the RAS oncoprotein

 The oncogenic mutations of RAS genes impair the intrinsic GTPase

activity of RAS that normally serves to turn its activity off, ensuring
that active signal transmission is transient.
The oncogenic mutations affecting ras genes
compromise Ras GTPase activity
 PTEN phosphatase, which counteracts PI3-
kinase by degrading its product,
phosphatidylinositol (3,4,5) trisphosphate
(PIP3).

 Loss-of-function mutations in PTEN

amplify PI3K signaling and promote
tumorigenesis in a variety of experimental
models of cancer
• mTOR kinase, a coordinator of cell
growth and metabolism that lies
both upstream and downstream of
the PI3K pathway.

• mTOR activation results, via

negative feedback, in the inhibition
of PI3K signaling.
• when mTOR is inhibited (rapamycin) in such cancer cells, the
associated loss of negative feedback results in increased activity of
PI3K and its effector Akt/PKB, thereby blunting the anti-proliferative
effects of mTOR inhibition.
1.3 Excessive Proliferative Signaling Can Trigger
Cell Senescence
 Excessively elevated signaling by oncoproteins, such as RAS, MYC, and RAF
in a normal cell provoke protective response such as induction of cell
death.
 Alternatively, cancer cells expressing high levels of these oncoproteins may
be forced to enter into the nonproliferative but viable state called
senescence.
 Whenever these tumor cells get the favorable microenvironment the enter
into proliferative phase.
• For example,

Cultured cells expressing high levels of the Ras oncoprotein may enter
into the non-proliferative but viable state called senescence; in
contrast, cells expressing lower levels of this protein may avoid
senescence and proliferate.
• Cells with morphological features of senescence, including enlarged
cytoplasm, the absence of proliferation markers, and expression of
the senescence-induced b-galactosidase enzyme, are abundant in the
tissues of mice engineered to overexpress certain oncogenes
2. Evading Growth Suppressors:

 Cancer cells must also circumvent powerful programs that negatively

regulate cell proliferation

 Depend on the actions of tumor suppressor genes

Growth suppressors are acting as the break mechanism to overrule

the initiation or “turning off” of cell division.
The two prototypical tumor suppressor genes encode the retinoblastoma (RB)-
associated and P53 proteins.
 The RB tumor suppressor protein limits cell
proliferation by preventing entry into the S phase of
the cell cycle

 The RB protein integrates signals from diverse

extracellular and intracellular sources and, in
response, decides whether or not a cell should
proceed through its growth and division cycle.
 • TP53 receives inputs from stress and
abnormality sensors that function within the
cell’s intracellular operating systems.

• If the degree of damage to the genome is

excessive, or if the levels of nucleotide pools,
growth-promoting signals, glucose, or
oxygenation are suboptimal, TP53 can call a
halt to further cell-cycle progression
Find examples of the loss of RB and TP53 protein and its impact on the cell
proliferation
Mechanisms of Contact Inhibition and Its
Evasion
• Healthy cell stops dividing when comes in contact with other cells but
cancer cell does not.

• Contact inhibition is an in vitro surrogate of a mechanism that operates

in vivo to ensure normal tissue homeostasis,
• Merlin, the cytoplasmic NF2 gene
product, activate contact inhibition
by coupling cell-surface adhesion
molecules like E-cadherin to
transmembrane receptor tyrosine
kinases.
• Merlin strengthens the adhesiveness of
cadherin-mediated cell-to-cell attachments
and thus inhibits the mitogenic signals.

• Thus, the mutation of NF-2 gene results in loss

of this property and thus grow in uncontrolled
manner.
 LKB1 epithelial polarity protein:

 Organizes epithelial structure and helps

maintain tissue integrity.

 LKB1 expression is suppressed, epithelial

integrity is destabilized, and epithelial
cells become susceptible

LKB1 has also been identified as a tumor suppressor gene that is lost in
certain human malignancies
Role of TGF-β
TGF- is best recognised for its anti-proliferative properties and cancer
cells' ability to evade it.

 TGF-b signalling is frequently misdirected in late-stage cancers away

from reducing cell proliferation and is discovered to activate a cellular
mechanism known as the epithelial-to-mesenchymal transition
(EMT),
3. Resisting Cell Death:
 The idea that apoptosis, or programmed cell death, acts as a natural
check against the development of cancer

 A mechanism known as apoptosis is normally used to instruct cells

to die when they get old or damaged.

 However, cancer cells manage to avoid regular cell death and keep
accumulating in the body.
Cancer cells acquires anti apoptotic regulators:-
• Tumor cells develops a variety of strategies to escape
apoptosis.

• Most common is the loss of P53 tumor suppressor

function,

• Alternatively, tumors may escape apoptosis by

increasing the expression of anti-apoptotic
regulators (Bcl-2, Bcl-XL Mcl-1)
• Anti-apoptotic proteins (e.g, BCL-2, BCL-XL,
MCL-1, BFL-1, BCL-W, and BCL2L10) preserve
outer mitochondrial membrane integrity by
inhibiting the pro-apoptotic members

• By downregulating pro-apoptotic Bcl-2–

related factors (Bax, Bim,Apaf-1 ).
Autophagy Mediates Both Tumor Cell Survival
and Death –
• Severely stressed cancer cells have
been shown to shrink via
autophagy to a state of reversible
dormancy.

• This particular survival response may

enable the cancer cells to survive
during anticancer therapy or during
shortage of nutrition.
Necrosis: Death of body tissue
• Necrosis has pro-inflammatory & tumor promoting potential

• Necrotic cells can release bioactive regulatory factors which can

directly stimulate viable neighbouring cells to proliferate

• More important, necrotic cell death releases pro-inflammatory

signals into the surrounding tissue microenvironment, in contrast to
apoptosis and autophagy, which do not.
Let's discuss about how cells die.
4. Enabling Replicative Immortality
Healthy cells self limit their replication
• In normal cell division, a small portion of
the end of each chromosome called
telomere, is lost every time DNA is copied.

• Loss of telomere reaches a critical point

and cell will no longer divide and replicate
and undergo p53 dependent cell cycle
arrest or apoptosis.
• But in cancer cells activation of an

enzyme called telomerase can maintain

telomeres and allow cells to replicate

limitlessly.
  The telomeres, composed of multiple
tandem hexanucleotide repeats,

 Eventually losing the ability to protect the

ends of chromosomal DNAs from end-to-
end fusions;

 Such fusions generate unstable dicentric

chromosomes that results in a scrambling
of karyotype that threatens cell viability.
Figure: Improved mode of the human telomeres

Jonathan B. Chaires, 2014, PLoS ONE 9(12): e115580.

 Telomerase, the specialized DNA polymerase that adds telomere
repeat segments to the ends of telomeric DNA

 Absent in non-immortalized cells but expressed at functionally

significant levels in the vast majority (>90%) of spontaneously
immortalized cells, including human cancer cells.
How cancer cell proliferate

 Cancer cells often avoid senescence or cell death by maintaining

their telomeres despite repeated cell divisions,

 Cancer cells achieve proliferative immortality by activating or

upregulating the normally silent human TERT gene (hTERT)

 hTERT encodes telomerase, a protein with reverse transcriptase

activity
hTERT which adds genetic units onto
the telomeres to prevent them from
shortening to the point of causing
senescence or cell death
 What is Quiescent cells and Senescent cells ?

Quiescence occurs due to lack of nutrition and growth

factors whereas senescence takes place due to aging and
serious DNA damages

Quiescent cells can re-enter the cell cycle, while

Senescent cells cannot re-enter the cell cycle
Quiescent cells:

• Quiescence is a reversible G0 state

• Therefore, the cells that reside in

quiescence are quiescent cells.

• Quiescent cells are in an inactive stage.

• Cells enter into the quiescent state due to

lack of nutrition and growth factors.
Quiescent Cells:

 Quiescent cells are characterized by a low RNA content, lack of cell

proliferation markers and increased label retention, indicating low cell
turnover.

 Quiescent cells are at rest.

 They can be activated and re-enter the cell cycle.

 Quiescence is beneficial, and it delays stem cell ageing.

Senescent cells:
• Senescence is a state of stable cell cycle arrest.

• senescence is a G0 state which is irreversible

• Senescence occurs due to ageing and serious DNA damage.

• These cells cannot re-enter the cell cycle. Senescent cells can no
longer replicate. Moreover, senescence is a degenerative process.
Senescent cells:

 Though senescent cells stop dividing, cells remain viable and

metabolically active for a certain period of time.

 Since senescent prevent replication, it serves an important anti-

tumorigenic function.
Similarity between the Quiescent cells and
senescent cells
• Both cells are two states of stable cell cycle arrest

• Both cells are in G0 state, which is an inactive state

• Therefore, both types of cells stop actively dividing

• Hence, both cells remain viable and metabolically active

Differences between the Quiescent cells
and senescent cells
5. Inducing Angiogenesis
• The formation of new blood vessels out of pre-existing capillaries.

ANGIOGENIC SWITCH OF TUMORS INVOLVES :

• Sprouting
• Splitting
• Remodeling of the existing vessels

WHY IT IS IMPORTANT?
Supply of oxygen and nutrients
Removal of waste products
TUMOR ANGIOGENESIS
• Three major steps

(A) Initiation of the angiogenic response.

(B) Endothelial cell(EC) migration, proliferation and tube formation.
(C) Finally the maturation of the neovasculature.
(A) Initiation of the angiogenic response.

• The first step in the formation of a capillary sprout from a pre-existing

mature blood vessel. (Proangiogenic growth factors secreted by the tumor
cell population)

• Proteolytic enzymes such as MMPs,Cathepsins etc

• localized degradation of the surrounding basement membrane

(B) Endothelial cell (EC) migration, proliferation and
tube formation.

• Next step is stimulus directed migration of ECs towards tumor mass

emanating from tumor itself

• Followed by division of ECs and lengthening the stalk of ECs sprout

• Lumen formation occurs with completion of capillary sprouts and loops

 (C) Finally the maturation of the neovasculature
Normal Tumor
 Single layer of peri-endothelial smooth muscle
cells that wraps around the endothelial cells are
known as pericytes.
 This is Critical for the development of a new
mature vascular network.
 It provide mechanical support, stability,
regulate the diameter of vessel and vascular
permeability
Pericytes
Pericytes are Important Components of the
Tumor Neovasculature-

• They provide important mechanical and

physiologic support to the endothelial cells

• Pericyte also secrete PDGF which helps

recruit pericytes and smooth muscle cells.
 A Variety of Bone Marrow-Derived Cells Contribute to
tumor Angiogenesis-

 These include cells of the innate immune system including

macrophages, neutrophils, mast cells, and myeloid progenitor.

 This tumor associated inflammatory cells can help to trigger the

angiogenic switch by providing tumor microenvironment and secrete
various growth factors
Angiogenic switch
• Epigenetic induction
• Genetic induction
Hypoxia, low Ph,
Mutant p53, Activated
cytokines (IL-6), bFGF,
oncogenes
Sex hormones

VEGF A VEGF A

VEGFR-1 VEGFR-2 VEGFR-3 Lymphogenesis

Vascular permeability, Migration, Mobilization, Proliferation, Survival

 Steps in activating invasion and metastasis

 Tumor suppressor genes mutation

Metastasis occurs  Proto-oncogene mutations: Group of genes

through a series of that cause normal cells to become cancerous
steps: when they are mutated
 local invasion,
 Intravasation,
 Transport,  Enhanced protease activity (e.g. MMPs)
 Extravasation,  Enhanced cell motility / interaction with
surrounding tissue.
 Colonization
 Decreased cell to cell adhesion and contact(
e.g. E-cadherin loss)
Steps in activating invasion and metastasis
• Intravasation and transport
through BM:

 Intravasation through BM
into blood vessel.

Interaction with vascular cells

Survival in circulation /
immune eversion
• Arrest and extravasation at secondary
site
Tumor cells interact with vascular cells.

• Invasion into secondary tissue and

formation of micro or macrometastasis
1) Interaction and adaptation to tissue
microenvironment.
2) Establishment of new vasculature.
3) Secondary tumor establishment or
dormancy.
Epithelial–mesenchymal
transition (EMT) play a critical
role in promoting metastasis in
epithelium-derived carcinoma
Epithelial to mesenchymal transition
program
• Loss of epithelial characteristics

• Attainment of mesenchymal markers

• Dynamic, multifaceted and often reversible

process

• Switch in intermediate filament usage

• Actin filament - EMT rearrangement

Resistance to anoikis (is a form of programmed cell death)

Phenotypic switch triggered by multiple components

 Where is EMT observed

1. EMBRYONIC
2. WOUND HEALING 3. CANCER METASTASIS
DEVELOPMENT
 Fibroblasts generation  Cancer progression via
 Gastrulation, neural crest
following tissue injury cell’s transition
cell migration, and
 Attenuation of  Incomplete EMT
 organ development
inflammation
 Organ fibrosis
Epithelial cells Mesenchymal cells
• Regular Columnar Morphology • Irregular round or elongate morphology
• Increased cell adhesion • Loss of polarity
• Presence of cell-cell junctions • Reduced cell–cell adhesive properties
• Relatively static cells • Cytoskeleton modulation
• Dynamic adhesions
• Lamellipodia and filopodia
• Highly mobile cells
Yang, J., Antin, P., Berx, G. et al. Guidelines and definitions for research on epithelial–
mesenchymal transition. Nat Rev Mol Cell Biol 21, 341–352 (2020).
1. Loss of Tight Junctions, Adherens Junctions and
Desmosomes

• Cell-cell contact disassembly

• Cytoskeletal protein redistribution
• Polarity disruption

2. Cytoskeletal Changes

• Actin stress fibers formed

• Anchor FACs for cell migration process
3. Transcriptional Shift

● Epithelial genes suppressed, mesenchymal genes activated -

EMT-TFs

● Vimentin upregulation , fibronectin deposition

4. Increased Migration and Motility

● N-Cadherin upregulation

● MMP secretion

● Integrin stimulation by ECM proteins

Biological Mechanisms in EMT
• INCREASED PROTEINS
● N-cadherin
● Vimentin
● Fibronectin
● Snail 1 (Snail)
● Snail 2 (Slug)
● Twist
● FOX C2
● SOX 10
● ZEB1
● MMP-2/3/9
● N-cadherin
Biological Mechanisms in EMT
• DECREASED PROTEINS
● E-cadherin - CDH1
● Desmoplakin
● Cytokeratin
● Occludin
• EXTRACELLULAR PATHWAYS
● TGFβ
● NF-kB
● Wnt/ß-catenin signaling

OTHER FACTORS
● ECM components
● IL-6 and ● Tumor Associated macrophages
 EMT in Cancer:
• METASTASIS:

 Cell invasion into Extracellular matrix

 Cytoskeletal reorganization, altered expression of cell adhesion

molecules

 Degradation of BM via MMP-2 and MMP-9 activation

 EMT in Cancer:
• TUMOR ANGIOGENESIS
• Expression of VEGF and EGF levels
• Reduced E-cadherin
• Notch and VEGF pathways cross talk – hypoxic tumors promote EMT

• IMMUNE ESCAPE
• High EMT Score, immune checkpoints presence
 EMT in Cancer:
• CHEMORESISTANCE
• Multidrug resistance, radioresistance
• Dysregulation of particular Transcriptional Factors
• Link between Cancer stem cells (CSCs) and therapy resistance
Live fluorescence microscopy imaging of metastasis — the
spread of cancer — showing melanoma cells (green) invading
lymphatic vessels (thick red vessels). Credit: W. KILARSKI, A.
LUND/EPFL
Energy Metabolism
Reprogramming Energy Metabolism
• Cancer metabolism is different than normal tissue metabolism.

• First time it was noted in 1920 by Biochemist Otto Warburg that

when cancer cells are provided with glucose, they generate large
amount of lactate regardless of whether oxygen is present or not.

• This metabolic difference is referred as THE WARBURG EFFECT……

 The normal cells utilize aerobic
respiration to completely catabolize of
glucose and generate cellular energy.

 Cancer cells rely primarily on glycolysis for

their metabolism to make lactate and it is
called aerobic glycolysis.
 ~18 fold lower efficiency of ATP
production relative to glycolysis and
mitochondrial oxidative phosphorylation

 However, cancer cells upregulated the

GLUT1, glucose transporter, that increase
glucose import into cytoplasm
 Glycolysis is activated under
the hypoxia condition

 That activate the HIF1α and

HIF2α

 It upregulate the glucose

transporter and multiple
enzymes of glycolysis pathway
 In many cancer types glucose uptake was
seen using Positron Emission Tomography
(PET) with radiolabelled analog

 18F-fluorodeoxuribose, FDG
 Cancer cells metabolize glucose for purpose
other than generating ATPs.

 Increase glycolysis allows the diversion of

glycolytic intermediates into various
biosynthetic pathways i.e. nucleoside and
amino acid synthesis

 Warburg like metabolism seems to be

presenting in rapidly dividing embryonic cells.
 Lactate produced from aerobic glycolysis
causes acidification of tumor cell which has
been shown to promote invasion and
metastasis.

 Lactate can also act as a nutrient for some

cells in the tumor.
Aerobic gylcolysis also generates ATP but less than aerobic

respiration.

 Cancer cells metabolize glucose for purpose other than generating

ATPs.

 Lactate produced from aerobic glycolysis causes acidification of

tumor cell which has been shown to promote invasion and

metastasis.

 Lactate can also act as a nutrient for some cells in the tumor.
What happen to lactate in cancer cells?

 In cancer, lactate plays an important role in angiogenesis

stimulating VEGF protein expression in endothelial cells.
 Lactate can enter tumor endothelial cells and lactate
released from tumor cells through MCT4 ( proton-coupled
lactate transporter) is enough to stimulate angiogenesis and
tumor growth
Glucose

Cancer Cells Surrounding Cancer Cells

Lactate

Part of Citric Acid Cycle

Mutation in metabolic enzymes causing
cancer
 Mutation in IDH [ isocitrate dehydrogenase]– In glioma, glioblastoma,
AML, myelodysplastic syndrome,ALL, prostate, colorectal cancer.

FH[fumarate hydratase]– metabolizes fumarate in TCA…mutation leads

to cancer
Targeting metabolism to treat carcinoma

 Folate– it can enhance cell proliferation. So antifolate is used as

chemotherapy.

 Metformin– recently two studies have shown that cancer related

mortality is decreased with metformin use - may be toxic to the cancer
Evading Immune Destruction
 The cells and tissues are constantly
monitored by an immune system,

 Immune surveillance is responsible for

recognition and elimination of cancer cells.

 Solid tumours that do appear have somehow

managed to avoid detection and thereby
evading eradications
Evading Immune Destruction
How tumor cells escape immune
recognition and destruction:

1) Low immunogenicity of tumor cells

 Failure to produce tumor antigen

 Mutation in MHC gene needed for
antigen processing.
 Inability to recognize tumor cells by
immune system.
Evading Immune Destruction
2) Tumor induced immune suppression-
 Factors secreted by tumor cells eg. TGF-b
inhibit T cells directly.

 TGF-β inhibits the development,

proliferation, and activation of immune
cells including T cells (CD4+ effector T cells
and CD8+ cytotoxic T cells), NK cells, and
macrophages.
Evading Immune Destruction
3) Tumor induced privileged
site-
 Factors secreted by tumor
cells create a physical barrier
to the immune system.

 Physical barrier, including

tumor stroma and extracellular
matrix, that limits the
penetration
Evading Immune Destruction
4) Tumor treated as self
antigen-
 Tumor antigens are taken up
and presented by APCs in
absence of co-stimulation
taken as self antigens and
escape from immune
destruction.
Evading Immune Destruction
4) Tumor treated as self antigen-
 PD-1/PD-L1 pathway controls the
induction and maintenance of immune
tolerance within the tumor
microenvironment.
Tumor-Promoting Inflammation

 Virtually every tumor contains immune cells present at varying

densities.

 Such immune responses are largely thought to reflect an attempt by

the immune system to eradicate tumors

 It has been demonstrated that the inflammatory response caused by

tumours paradoxically promotes the development and spread of
tumours.
 Inflammation can support a variety of characteristic functions, By
delivering bioactive chemicals for a favourable tumour micro-
environment,
a. Growth factors that sustain proliferative signaling
b. Survival factors that limit cell death
c. Proangiogenic factors
d. Extracellular matrix-modifying enzymes that facilitate
angiogenesis, invasion and metastasis.
Additionaly, Reactive oxygen species that are actively mutagenic for
nearby cancer cells, accelerate their genetic evolution towards the
malignancies
• Recent study, suggest that immune system operates as a significant
barrier to tumor formation and progression,

• Example: It was observed that tumors arose more frequently or grew

more rapidly in the immuno-deficient mice. Particularly, deficiency in
the development of function of CD8+, CTL, DC4+, T helper cells, NK
cells
 Genome Instability and Mutation
 Genomic instability and mutation acts as enabling hallmark of cancer

 DNA damage or mutation in a normal cell results in cell cycle arrest

followed by DNA repair or apoptosis.

 Interference in this process may occur either by lack of recognizing

and repair of damaged DNA
Level of disruption types of gene instablity
1) Nucleotide instability:
Include nucleotide substitution, deletion or insertion
E.g. xeroderma pigmentosum

2) Microsatellite instability
Include defect in mismatch repair leads to contraction or expansion of
microsatellite
E.g. Lynch syndrome (hereditary colorectal (colon) cancer)
 Level of disruption types of gene instability
3) Chromosomal instability:

 Most prominent form

 90% of human cancer exhibiting chromosomal abnormalities. It include

chromosomal anuploidy, amplifications, deletions, translocations and
inversions.

Examples: Breast, prostate, non small cell lung cancer, leukaemia,

neuroblastoma etc.
Level of disruption types of gene instablity
 Defects in these caretaker genes:

 DNA damage and inactivation of repair machinery

CGH (comparative genomic hybridization)-

One method of molecular genetic analysis to compare patient DNA to

reference DNA to check the gains and losses of gene copies in the patients
cell genome by using florescent dye .
TUMOR MICROENVIRONMENT
TUMOR MICROENVIRONMENT

ECM= Extracellular matrix

T cells =
Adipocyte =
CSC = Cancer stem cells
Macrophages =
Neutrophil =
Fibroblasts =
Cancer-Associated Fibroblasts

 Cells with similarities to the fibroblasts that provides structure

support to most of normal epithelial tissues.

 In the tumour microenvironment, a subset of activated fibroblasts

known as cancer-associated fibroblasts (CAFs) exhibits substantial
variability.
CAFs play a key role in cancer progression by contributing to;

 Extracellular matrix (ECM) deposition and remodeling,

 Extensive crosstalk with cancer cells,

 Epithelial-to-mesenchymal transition (EMT),

 Invasion, metastasis, and therapy resistance

Cancer-associated fibroblasts (CAFs) constitutes at least two
distinct cell types:
1) Cells with similarities to the fibroblasts
that provides structural support to most
of normal epithelial tissues

2) Myofibroblasts, whose biologic roles

and properties differ markedly from those
of the widely distributed tissue-derived
fibroblasts.
Myofibroblasts:

 Myofibroblasts are identifiable by their expression of alpha-smooth

muscle actin (SMA).

 They are rare in most healthy epithelial tissues, although certain

tissues, such as the liver and pancreas,

 Myofibroblasts transiently increase in abundance in wounds and are

also found in sites of chronic inflammation.
Myofibroblasts:

• Myofibroblasts are problematic in chronic inflammation, contributing

to the pathological fibrosis observed in tissues such as lung, kidney,
and liver

• Have been demonstrated to enhance tumor phenotypes, notably

cancer cell proliferation, angiogenesis, and invasion and metastasis
Endothelial Cells

• Endothelial cells form a single cell layer

that lines all blood vessels and regulates
exchanges between the bloodstream and
the surrounding tissues.

• Forming the tumor-associated vasculature

 The angiogenic switch activates the quiescent endothelial cells, to
enter into a cell biological program.

 Endothelial cells have prominent VEGF, angiopoietin, FGF signal in

addition, Notch, Neuropilin, ROBO and Eph-A/B signals

 This will help to develop novel therapies.

Pericytes:

 Specialized mesenchymal cell type with

fingerlike projections that wrap around
the endothelial tubing of blood vessels.

 Provide supportive framework to

endothelial cells.
Pericytes:
• In normal tissues, pericytes are known to provide paracrine support
signals to the normally quiescent endothelium
• Pericytes also collaborate with the endothelial cells to synthesize the
vascular basement membrane that anchors both pericytes and
endothelial cells and helps vessel walls to withstand the hydrostatic
pressure of blood flow.

• supporting the tumor endothelium

Immune Inflammatory Cells

• Fighting against the infection and wound healing, the immune cells
transiently appear and disappear

• these cells are associated with the various tissues such as fibrosis,
angiogenesis, and neoplasia

• So, tumor promoting cells now includes;

Immune Inflammatory Cells
 So, tumor promoting cells now includes; Macrophage subtypes, mast
cells, neutrophils as well as T and B lymphocytes

 Signalling molecules by inflammatory cells that serve as effector of

their tumor promoting actions such as

 EGF, VEGF-A/C, FGF2, chemokines, cytokines , MMP-9, cysteine

cathepsin proteases etc.

 Facilitate tissue invasion, and to support the metastatic

dissemination and seeding of cancer cells.
Stem and Progenitor Cells of the Tumor
Stroma
• Also known as cancer initiating cell.

• Cell type capable of initiating and sustaining growth of the tumor.

• These fraction of tumor cells were capable of tumor development

and have great proliferative within a tumor.
Stem and Progenitor Cells of the Tumor
Stroma
• Progenitor cells are descendants of stem cells that then further
differentiate to create specialized cell types

• The bone marrow has increasingly been implicated as a key source

of tumor associated stromal cells.

• Mesenchymal stem and proginator cells have been found to transit

into tumors from the marrow
Stem and Progenitor Cells of the Tumor
Stroma