Cellular signaling and cancer

By/ Dr.Asma Abdullwahab Khalifa

 Introduction / Overview of Cell
Communication
I. Local vs. Long Distance Signaling
 Cell to Cell Contact (ex: cell junctions & cell-cell recognition)
 Secretion of Local Regulators (local)
 Neurotransmitters and Neurons (local)
 Hormones traveling in blood (long distance)

III. 3 Stages of Cell Signaling

1 - Reception (of signal = ligand) & Receptors
* 3 receptors: (1) G-protein couple receptors
(2) Receptor Tyrosine
Kinases
(3) Ion Channel Receptors
2 – Transduction (a multistep pathway)
* Phospholylation by Kinases
* 2nd messengers – ex: cyclic AMP (cAMP) and
Calcium & IP3
3 – Cellular Response
* Cytoplasmic
* Nuclear
 Evolution of Cell Signaling

• A signal transduction pathway is a series of

steps by which a signal on a cell’s surface is
converted into a specific cellular response
• Signal transduction pathways convert signals
on a cell’s surface into cellular responses
 The Three Stages of Cell Signaling:

• The first research into cell signaling involved

how the hormone epinephrine acts on cells
• The cells receiving signals went through three
processes:
– Reception
– Transduction
– Response
EXTRACELLULAR CYTOPLASM
FLUID
Plasma membrane

1 Reception

Receptor

Signaling
molecule
EXTRACELLULAR CYTOPLASM
FLUID
Plasma membrane

1 Reception 2 Transduction

Receptor

Relay molecules in a signal transduction pathway

Signaling
molecule
EXTRACELLULAR CYTOPLASM
FLUID
Plasma membrane

1 Reception 2 Transduction 3 Response

Receptor
Activation
of cellular
response
Relay molecules in a signal transduction pathway

Signaling
molecule
Reception: A signal molecule binds to a
receptor protein, causing it to change
shape
• The binding between a signal molecule (ligand)
and receptor is highly specific
• A shape change in a receptor is often the initial
transduction of the signal
• Most signal receptors are plasma membrane
proteins
 Receptors in the Plasma Membrane

• Most water-soluble signal molecules bind to

specific sites on receptor proteins in the plasma
membrane
• There are three main types of membrane
receptors:
1. G protein-coupled receptors
2. Receptor tyrosine kinases
3. Ion channel receptors
• A G protein-coupled receptor is a plasma
membrane receptor that works with the help of
a G protein
• The G protein acts as an on/off switch: If GDP is
bound to the G protein, the G protein is inactive
 Signaling-molecule binding site

Segment that
interacts with
G proteins

G protein-coupled receptor
 G protein-coupled Plasma Inactive
membrane Activated Signaling molecule enzyme
receptor
receptor

GDP

G protein Enzyme GDP GTP

CYTOPLASM (inactive)

1 2

Activated
enzyme

GTP
GDP
Pi

Cellular response

3 4
• Receptor tyrosine kinases are membrane
receptors that attach phosphates to tyrosines
• A receptor tyrosine kinase can trigger multiple
signal transduction pathways at once

Copyright © 2008 Pearson Education, Inc., publishing as Pearson Benjamin Cummings

Signaling Ligand-binding site
molecule (ligand)
Signaling
molecule
 Helix

Tyr Tyr Tyr Tyr Tyr

Tyr
Tyrosines Tyr Tyr
Tyr Tyr Tyr Tyr
Tyr Tyr Tyr Tyr Tyr
Tyr

Receptor tyrosine
kinase proteins Dimer
CYTOPLASM

1 2

Activated relay
proteins

Cellular
Tyr Tyr P Tyr Tyr P Tyr Tyr P response 1
P
Tyr Tyr P Tyr Tyr P P Tyr Tyr P
Tyr Tyr P Tyr Tyr P P Tyr Tyr P Cellular
6 ATP 6 ADP response 2

Activated tyrosine Fully activated receptor

kinase regions tyrosine kinase
Inactive
relay proteins

3 4
• A ligand-gated ion channel receptor acts as a
gate when the receptor changes shape
• When a signal molecule binds as a ligand to the
receptor, the gate allows specific ions, such as
Na+ or Ca2+, through a channel in the receptor
1 Signaling
Gate
molecule closed Ions
(ligand)

Plasma
Ligand-gated membrane
ion channel receptor

2 Gate open

Cellular
response

3 Gate closed
 Transduction: Cascades of molecular
interactions relay signals from receptors
to target molecules in the cell
• Signal transduction usually involves multiple
steps
• Multistep pathways can amplify a signal: A few
molecules can produce a large cellular response
• Multistep pathways provide more opportunities
for coordination and regulation of the cellular
response
 Protein Phosphorylation and
Dephosphorylation
• In many pathways, the signal is transmitted by a
cascade of protein phosphorylations
• Protein kinases transfer phosphates from ATP to
protein, a process called phosphorylation
Signaling molecule

Receptor
Activated relay
molecule

Inactive
protein kinase
1 Active
protein
kinase
1
Inactive
protein kinase ATP
ADP Active P
2
protein
PP kinase
Pi 2
Inactive
protein kinase ATP
ADP Active P
3
protein
PP kinase
Pi 3
Inactive
protein ATP
ADP P
Active Cellular
protein response
PP
Pi
 Small Molecules and Ions as Second
Messengers
• The extracellular signal molecule that binds to
the receptor is a pathway’s “first messenger”
• Second messengers are small, nonprotein,
water-soluble molecules or ions that spread
throughout a cell by diffusion
• Cyclic AMP and calcium ions are common
second messengers
• Second messengers participate in pathways
initiated by G protein-coupled receptors and
receptor tyrosine kinases
 Adenylyl cyclase Phosphodiesterase

Pyrophosphate
P Pi

ATP cAMP AMP

 First messenger

Adenylyl
G protein cyclase

G protein-coupled GTP
receptor

ATP
Second
cAMP messenger

Protein
kinase A

Cellular responses
EXTRACELLULAR
Plasma
FLUID
membrane

Ca2+ pump
ATP
Mitochondrion

Nucleus

CYTOSOL

Ca2+
pump
Endoplasmic
reticulum (ER)
Ca2+
ATP pump

Key

High [Ca2+]
Low [Ca2+]
• A signal relayed by a signal transduction
pathway may trigger an increase in calcium in
the cytosol
• Pathways leading to the release of calcium
involve inositol triphosphate (IP3) and
diacylglycerol (DAG) as additional second
messengers
EXTRA-
CELLULAR Signaling molecule
FLUID (first messenger)

G protein

DAG

GTP
G protein-coupled PIP2
receptor Phospholipase C
IP3
(second messenger)

IP3-gated
calcium channel

Endoplasmic
reticulum (ER) Ca2+

CYTOSOL
EXTRA-
CELLULAR Signaling molecule
FLUID (first messenger)

G protein

DAG

GTP
G protein-coupled PIP2
receptor Phospholipase C
IP3
(second messenger)

IP3-gated
calcium channel

Endoplasmic
reticulum (ER) Ca2+

Ca2+
(second
CYTOSOL messenger)
EXTRA-
CELLULAR Signaling molecule
FLUID (first messenger)

G protein

DAG

GTP
G protein-coupled PIP2
receptor Phospholipase C
IP3
(second messenger)

IP3-gated
calcium channel

Endoplasmic Various
reticulum (ER) Cellular
Ca2+ proteins
responses
activated
Ca2+
(second
CYTOSOL messenger)
 Response: Cell signaling leads to
regulation of transcription or
cytoplasmic activities
• The cell’s response to an extracellular signal is
sometimes called the “output response”
• Ultimately, a signal transduction pathway leads
to regulation of one or more cellular activities
• The response may occur in the cytoplasm or
may involve action in the nucleus
• Signaling pathways can also affect the physical
characteristics of a cell, for example, cell shape
 Nuclear and Cytoplasmic Responses

• Many signaling pathways regulate the synthesis

of enzymes or other proteins, usually by turning
genes on or off in the nucleus
• The final activated molecule may function as a
transcription factor
• Other pathways regulate the activity of enzymes
 Growth factor
Reception
Receptor

Phosphorylation
cascade
Transduction

CYTOPLASM

Inactive Active
transcription transcription
factor factor
Response
P

DNA

Gene

NUCLEUS mRNA
 Signal Amplification

• Enzyme cascades amplify the cell’s response

• At each step, the number of activated products
is much greater than in the preceding step
Reception

Binding of epinephrine to G protein-coupled receptor (1 molecule)

Transduction
Inactive G protein
Active G protein (102 molecules)

Inactive adenylyl cyclase

Active adenylyl cyclase (102)

ATP
Cyclic AMP (104)

Inactive protein kinase A

Active protein kinase A (104)

Inactive phosphorylase kinase

Active phosphorylase kinase (105)

Inactive glycogen phosphorylase

Active glycogen phosphorylase (106)

Response
Glycogen
Glucose-1-phosphate
(108 molecules)
 Cell signaling summary
• Ligand
• Receptor
• Adapters or enzymes
• Signal cascades Change in cell phenotype:
• Transcription factors change in cell behavior

 Cancer is really a disease of an aberrant signal

processing.
 Oncoproteins and tumor suppressors create cancer
through their ability to generate signaling imbalance.
 Cancer is a disease of uncontrolled cell proliferation.
• Receptor tyrosine kinase(RTKs), which have
tyrosine amino acids attach to the intracellular
catalytic sections involved in the phosphorylation
of a small set of intracellular proteins. Most of the
growth factor families belong to receptor tyrosine
kinase, including epidermal growth factor receptor
family: EGFR (ErbB- 1), HER2/neu (ErbB- 2), HER3
(ErbB- 3), and HER4 (ErbB- 4); Fibroblast growth
factor receptor (FGFR) family: comprised of 23
members to become the largest family of growth
factor; vascular endothelial growth factor receptor
family; Ephrin receptor family and RET receptor
family
• RTKs have been shown not only to be the key
regulators of normal cellular processes but also to
have a critical role in the development and
progression of many types of cancer. These
receptors are involved in the regulation of cell
growth, proliferation, differentiation and survival.
RTKs have the ability to activate intracellular protein
by phosphorylation, triggering the downstream
signalling.
• RTKs act in pairs.
• When a signal molecule binds to an RTK, this one and the
neighbouring RTK associate with each other forming a
cross- linked dimer.
• Each RTK in the dimer phosphorylates the tyrosines on the
other RTK. This process is called cross- phosphorylation.
• Once cross- phosphorylated, the intracellular enzymatic
domain of the RTKs serve as docking platform for different
intracellular proteins that have SH2 domain to bind to the
phosphorylated tyrosines. This process can cause multiple
different SH2- containing proteins to bind at the same time
to any of these phosphorylated domanis thus allowing the
activation of multiple intracellular signalling at the same
time.
The major cancer- related signalling pathways/ Ligands or other activating functions
in blue and receptors in red. Pathways and activated functions in green. 1: Ras
network (A: Ral pathway, B: MAPK pathway, C: PI3K/ AKT pathway). 2: mTOR
pathway. 3: Jack- Stat pathway. 4: WNT pathways. 5: cAMP pathway. 6: Focal
adhesion pathway. 7: Tgf- beta pathway. 8: Hedgehog pathway. 9: NfKb pathway.
Note the cross talking between all the pathways.
 The Ras Network
• The Ras protein family, composed by N, H, and
K Ras, control a large signalling network
widely involved in cancer.
• Three main pathways have been identified:
Ras Raf Erk (MAP kinases), Ras PI3K AKT/ Pkb,
and Ras Ral Gef.
• Upstream to the Ras proteins is a family of
RTKs mostly binding to growth factors.
 Raf Erk (MAP kinases)
• The mitogenic activated protein kinases pathway is highly
conserved and regulates mainly proliferation and differentiation.
It also controls, with the PI3K pathway, the Retinoblastoma
suppressor gene activity.
• Mechanisms:
Once activated, the Ras proteins interact with the proteins
of the Raf family (A- Raf, B- Raf, and C- Raf) which in turn activated
Mek and then the Erk scaffolding proteins (i.e. Raf1, Mek 1,2, Mp1,
and Erk). ‘Scaffolding protein’ is defined as a protein able to interact
with several different members of a pathway forming multiprotein
complexes. The ‘Erk scaffolding system’ eventually activates the
intranuclear targets (Elk1, cMyc, CyclinD1) acting on proliferation
and differentiation . According to the type of cells either a positive
or a negative loop can be triggered leading to further proliferations
or self limitation.
• Involvement in cancer:
Persistent high levels of ligands in the extracellular
environment, mostly growth factors, and
activating mutations, mostly in the H- , K- , and N-
Ras, B- Raf, C- Raf, or Mek1/ 2 genes, are the main
alterations found in cancer which lead to
abnormal signalling of this pathway. This set of
mutated proteins is becoming an attractive target
for numerous drugs
 PI3K AKT/ Pkb
• The phosphatidylinositol 3’– kinase (PI3K)- AKT
pathway is one of the most crucial in cell
physiology and commonly involved in cancer as it
is at the center of a large network regulating
several vital functions of the cell.
• It is activated by several types of cellular stimuli
or external toxic insults and regulates
transcription of genes and translation into
proteins, cell cycle, and cell survival.
• It is under control, through the Ras family, of RTK
or, through the cAMP pathway of the GPCR.
 Mechanisms
• PI3K enzymes are divide into three classes: I, II, and III.
• Class I are dimers formed by a regulatory and a catalytic subunits which, in normal
cells, are activated by either growth factors, through RTKs receptors, GPCRs and
Ras proteins.
• Once activated, Ras stimulates class Ia and Ib PI3K isoforms respectively.
• PI3K catalyses the production of phosphatidylinositol-3,4,5- triphosphate (PIP3) at
the cell membrane.
• PIP3 in turn serves as a second messenger that helps to activate AKT.
• This regulation is also mediated by Pdk1, a positive regulator, and Pten that has an
inhibitory effect.
• Once active, AKT phosphorylates more than 200 substrates controlling key cellular
processes by phosphorylating substrates involved in cell survival, motility, growth,
glycolysis, cell cycle, and DNA repair.
• It also interacts with mTOR pathway which in turn controls cell survival, motility,
microtubules lipids, autophagy, and protein synthesis.
• Class II PI3K are monomers mainly involved in regulating
vesicular traffic including the transport on the cell membrane
of the glucose transporter Glut4.
• PI3K- C2alpha is the member of this class for which we have
more information and, alongside Glut4, is also involved in
endocytosis and inducing sprouting angiogenesis.
• Another member of this class, PI3K- C2beta, less well
investigated, appears to be involved instead in cell migration
and inhibition of apoptosis.
• Both class I and class II are downstream to RTKs and GPCRs.
• Finally, Class III PI3K are involved in vesicular trafficking and
nutrient sensing
• PI3K/ AKT pathway also exercises its physiological regulatory
roles through epigenetic mechanisms as AKT phosphorylates
substrates that regulate chromatin conformation. Because of
this epigenetic modification, the activity of these substrates is
altered, making the DNA available, or not for transcription.
 Involvement in cancer
• This pathway has a unique role in cancer, not only because of the variety of cellular
functions affected but also because, unlike other pathways, every major hub of this
pathway can be altered in cancer.
• Class Ia PI3K includes four catalytic subunits (p110 alpha, beta, gamma, and delta)
and one regulatory subunit p85, which are involved by genetic damage. Mutations
are found in p110 alpha, beta, delta and p85. Amplification is instead found in p110
alpha, beta, gamma, and delta.
• Pten is a suppressor gene as it physiologically blocks AKT activation, and like all
suppressor genes it can be also altered in hereditary tumours.
• AKT three members (1, 2 and 3) can be affected by mutation (AKT3) and
amplification (AKT1 and AKT2).
• As the Pik3 pathway is also involved in epigenetic regulation, alterations to its
activity can lead to epigenetic changes which can promote oncogenesis. An
example is the ability of AKT to reduce DNA replication- associated DNA
methylation, increasing transcriptional activity. AKT stabilizes, through
phosphorylations, DNA methyltransferase 1 (DNMT1), which in turn prevents DNA
methylation by other enzymes. As a result, transcription increases.
• By phosphorylating the histone Ezh2, AKT diminishes the affinity of this histone for
chromatin thus reducing its transcription- repressing activity.
 Ral (Ras-like small GTPases)
• This is a small collateral pathway which is divided
into two: through RalA is involved in control of
motility, glycolysis, and interact with the MAPK
pathway. It is also connected with the Jack/ Stat
pathway to activate Jak. RalB controls endocytosis
and apoptosis through direct binding to Exo84 and
inducing downstream activation of ULK1 and
Beclin1-VPS34 complex.
• Both branches affect transcription, mitophagy, and
autophagy.
• Involvement in cancer. Ral is very much involved in
oncogenesis, although our knowledge is still
sketchy. Ralbp1 is a large protein with several
binding domains and is a hub involved in
regulation of endocytosis, motility, autophagy, and
metabolism regulation.
• Glycolysis is regulated by delivering phopso-Drp1
to mitochondria.
• Exo84 and Sec5 are other crucial hubs involved in
transcription
 The mTOR pathway
• mTOR (mammalian Target Of Rapamycin) pathway is a hub nested
between other pathways and at its core are two super complexes: the
mTorc1 and the mTorc2 which have serine/ threonine protein kinase
activity.
• It owes its unusual name to the fact that this pathway was discovered
to be the target of Rapamycin, an antifungal drug. Both super
complexes are mainly controlled by the PI3K pathway, through the
Tsc1/ 2- Tcb1D7 intersection mTorc1 and directly, mTorc2.
• However, the Tsc1/ 2- Tcb1D7 intersection mTorc1 is also regulated by
the Wnt, the Mapk, the AMPK, and the hypoxia pathways. When
active mTorc1 regulates, through transcription of target genes, a
series of functions, including cell proliferation and macromolecules
expression. It also inhibits mTorc2. mTorc 2 instead acts as a loop and
sends regulatory signals back to the PI3K, the Wnt, and the cAMP
pathways, eventually affecting cell survival, glucose metabolism, and
motility.
 Mechanisms
• Activation of PI3K and Mapk is a key event to induce proliferation, however it is
essential that a cell does not proliferate if certain conditions, like availability of
nutrients, adequate energy metabolism, and molecular building blocks, are not
available.
• mTor is a hub which provides coordination between proliferative signals and the
presence of adequate intracellular environmental conditions .
• mTorc1 is a crucial hub which coordinates the presence of a proliferative signal with
the availability of necessary features like ATP, oxygen, glucose, and also amino acids
• In a normal cell, through the mTorc1, the proliferating signal can be overrun and
deactivated if the conditions are not right less proliferation will be achieved.
• If intracellular conditions are adequate and mTorc1 is activated, it will not only
allow proliferation but will also stimulate mitochondria and glycolysis to maintain
optimal ATP concentration, protein synthesis, and lipid synthesis to provide the
building blocks for mitosis
• mTorc2 through the Sgk1 genes and, again the PI3K/ AKT pathway, it is also
involved in controlling apoptosis and glucose metabolism. Finally, through the
regulation of Wnt/ Ca2+ and cAPM pathways promotes cell motility and
cytoskeleton organization
 Involvement in cancer
• The mTOR pathway is often activated in tumours,
and its not only regulates gene transcription and
protein synthesis to regulate cell proliferation and
immune cell differentiation but also plays an
important role in tumur metabolism
• Therefore,, avariety of drugs have been proven to
have high activity in combination with mTOR
inhibitors
 Other patways
• The JAK- STAT pathway
• The WNT pathway
• The cAMP/ GPCR pathway
• The focal adhesion kinase pathway
• The other dual address pathways: NFKb, TGF-
beta, and hedgehog
 Summary
• Oncogenic mutations can cause the affected genes to be
overexpressed (e.g., gene amplification) or produce mutated
proteins whose activity is dysregulated (e.g., point mutations,
truncations, and fusions).
• Examples include proteins involved in signaling pathways that are
commonly activated in many physiological responses, such as
growth factor receptor tyrosine kinases (RTKs; e.g., the epidermal
growth factor receptor, EGFR), small GTPases (e.g.,Ras),
serine/threonine kinases (e.g., Raf and Akt), cytoplasmic tyrosine
kinases (e.g., Src and Abl), lipid kinases (e.g., phosphoinositide 3-
kinases, PI3Ks), as well as nuclear receptors (e.g., the estrogen
receptor, ER).
• Components of developmental signaling pathways, such as Wnt,
Hedgehog (Hh), Hippo, and Notch can also be affected, as can
downstream nuclear targets of signaling pathways—for example,
transcription factors (e.g.,Myc andNF-kB), chromatin remodelers
(e.g., EZH2), and cell cycle effectors (e.g.,cyclins).
• Alternatively, deletions and other mutations can inactivate
negative regulators that normally function as tumor
suppressors.
• Indeed, one of the most commonly mutated genes in cancer
is the tumor suppressor p53, the so-called “guardian of the
genome.”
• p53 is a critical molecule that controls cell proliferation and
stress signals such as apoptosis and DNA damage responses.
• pRB and CKIs such as p16 are other tumor suppressors whose
mutation deregulates the cell cycle.
• Many tumor suppressors function as negative regulators of
cytoplasmic signaling—for example, the adenomatous
polyposis coli protein (APC) is a negative regulator of the Wnt
pathway, and the lipid phosphatase PTEN is a negative
regulator of the PI3K-Akt pathway.