Cancer cells attain autonomy

Low reqmt for growth stimulatory molecules

No sensitivity to inhibitory signals from adjacent cells
The altered transcriptional activity - growth
Extracellular signals influence cell growth
ignalling molecules can be large or small
Large molecules are polypeptides!proteins
"eceptors-nucleus by ptn-ptn interaction!cytos#eletal changes and movmt of
macromolecules within intracellular compartments$
Low mwt cpds-lipid sol regulators-ex % retinoic acid !steriod hormones$
Categorisation of EC signals
&cc to source of signal
Endocrine- hormones from bd stream
&utocrine
paracrine
'rowth factor receptors
'rowth factor receptors can be divided to ( groups$
Tyr phosphorylation is less than )*
+inase receptor % those contain tyr #inase activity as an integral part of receptor
Those that can recruit a #inase as a result of ligand binding
"eceptors with integral #inase activity
Extracellular ligand binding domain
Transmembrane region
,ultifunctional cytoplasmic tail-&T- binding site plus tyrosine #inase activity
"eceptors have to be dimeric to be active
"eceptors that recruit tyrosine #inases
JAK-STAT . cyto#inesgrowth factors that stimulate components of the hematopoietic system$
These particular receptor proteins do not carry covalently associated tyrosine #inase
domains and instead form noncovalent complexes with tyrosine #inases of the
/a# 0/anus #inase1 class 02igure 3$((1 2ollowing ligand-mediated receptor dimeri4ation! the /a#
en4yme associated with each receptor molecule phosphorylates tyrosines on the cytoplasmic tails
of the partner receptor molecule! much li#e the transphosphorylation occurring
after ligand activation of receptors li#e the E'2-" and -5'2-"$ The resulting
phosphotyrosines attract and are bound by 6(-containing transcription factors
termed T&Ts 0signal transducers and activators of transcription1$ 7nce they are associated with
the receptors! these T&Ts become phosphorylated by the /a#s$ This creates individual T&T
molecules that possess both 6( groups and phosphotyro sines$ 8mportantly! the 6( groups
displayed by the T&Ts have a specificity for binding the phosphotyrosine residues that have just
been created on the T&Ts$ Consequently! T&T-T&T dimers form! in which each T&T
uses its 6( group to bind to the phosphotyrosine of its partner$ The resulting
dimeri4ed T&Ts migrate to the nucleus! where they function as transcription
factors 0see 2igure 3$((1$ The T&Ts activate target genes that are important for cell proliferation
and cell survival$ 8ncluded among the proliferation and survival genes are 8nyc! the genes
specifying cyclins 5( and 59 0which enable cells to advance through their! anti-apoptotic
protein :cl-;L$ re-engineering of the tat9 protein through the introduction of a pair of cysteine
residues! which causes the resulting mutant tat9 to dimeri4e spontaneously! forming stable
covalent disulfide cross-lin#ing bonds$ These stabili4ed tat9 dimers are structural and functional
mimics of the dimers that are formed normally when tat9 is phosphorylated by /a#s$ This mutant
tat9 protein is now constitutively activetat9 has been in particular found to be activated in
many melanomas!breast cancer and head and nec# cancers$
"eceptors that act via ser<thr #inase
:oth stimulatory and inhibitory signals can be relayed
The phosphorylated proteins and 6 domains do not provide the doc#ing mechanism
with other proteins
T'2-beta receptor has widespread functions- induction of EC, proteins! stimulation of
-5'2 and chemotaxis
&denylate cyclase-lin#ed receptors
"eceptors have a single protein chain loop across the membrane seven times
The cytoplasmic domain interacts with ( regulatory proteins to form the adenylate
cyclase complex
Ligand binding activates adenylcyclase and increases intracellular c&,-
'n"6 modulates ovarian steroid hormone function and is the polypeptide in these
receptors$
Integrin signalling
8ntegrins chiefly perform three functions
)$physically lin#ing cells to EC,
($8nforming cells whether or not tethering to certain EC, components has been
achieved$
9$2acilitating motility by ma#ing and brea#ing contacts with the EC,$
integrins may cluster and form multiple lin#s to the EC, in small! locali4ed areas termed focal
adhesions 0see 2igure =$(>&1 $ uch clustering provo#es activation of focal adhesion #inase
02&+1 ! a nonreceptor tyrosine #inase li#e rc! which is associated with the cytoplasmic tails of
the ? subunits of certain integrin molecules and becomes phosphorylated! presumably
by transphosphorylation! once integrins congregate in these focal adhesions$ 7ne of the re@lting
phosphotyrosine residues on 2&+ provides a doc#ing site for rc molecules 02igure 3$(A&1$ rc
then proceeds to phosphory tyrosine-phosphorylated 2&+! can recruit os into this complex$ The
latter proceeds to activate its normal downstream targets including "as and -89+$
Three important signaling pathways of Ras
'rowth factor elicits a multiple cellular repsonse$
The cognate receptor of the '2 is able to activate a specific combination of downstream
signalling pathways$
Exaggerated forms of this signalling operates in cancer cells that experiences continous
growth factor stimulation$
This could be due to autocrine signalling loop or a mutant or constitutively activated
receptor$
Bhen "& binds to 'T-!a domain of the "as protein ! termed its E22ECT7" L77- is
able to interact physically with several alternative downstream signalling partners that are
#nown as "as effectors$
Each of these effectors bind quite tightly to the effector loop of the 'T--bound "as
protein while having little affinity for the loop presented by its '5--bound form$
The first of these Ras effectors was the Raf kinase.
The activation of "af by "as depends on the relocalisation of "af within the cytoplasm$
"as proteins are always anchored to the inner surface of plama membrane$
7nce it binds 'T-!&ctivated "as attracts and binds "af via its effector loop$
-rior to this "af is present in cytosol$
&ssociation of "as-"af seems to shift the three dimensional configuration of the "af
#inase molecule$
5uring the time its is anchored to "as!"af may become phosphorylated by #inases$
"af now actively proceeds to phosphorylate activate ,E+ 0dual #inase1 which can
phosphorylate both tyr residues and ser-threonine residues$
,E+ phosphorylates E"+ $
E"+ phosphorylates substrates that in turn regulate various cellular processes inlcuding
transcription$0el#! ets! &-1
&mong the genes induced by the "af-,E+ %Er# pathway is the activation of the
promoters of the two immediate early genes encoding the 2os and /un transcription
factors$
7nce synthesised! these two proteins can associate with one another to form &--) !a
widely acting heterodimeric trancription factor that is often found in hyperactivated form
in cancer cells
8mportance . &ctivation of transcription factors
&ctivation of #inases
&ctivated #inases phosphorylate and reconfigure chromatin associated proteins ma#ing it
more conducible to transcription
&ctivates numerous growth promoting genes
Confers anchorage independence and loss of contact inhibition$
Contributes to the change in cell shape associated with transformation$
Second downstream pathway that Ras controls is the Akt/PK kinase
This pathway evo#es other cellular responses$
The most important is suppression of cellular apoptosis$
This anti-apoptotic effect is criticial for cancer cells
-hospholipids contain at their hydrophilic heads!an inositol group$
The inositols moiety of such phospholipids can be modified by the addition of phosphate
groups$
The resulting phosphoinositol may then be cleaved away from the remaining largely
hydrophobic portions of a phospholipid molecule$
ince it is purely hydrophilic !this phosphoinositol!termed 8-9 can then diffuse away
from the membrane $
8t thereby serving as an intracellular hormone to dispatch signals from the plasma
membrane to distant parts of the cell$
uch intracellular hormones are termed as Csecond messengersD
The second product of this cleavage!5&' serves to activate #ey sigalling #inase in the
cell-the serine threonine #inase #nown as protein #inase C0-+C1
'T- activated "as binds to -89+ and enhance functional activation
"as--89+ binding causes -89+ to be closely associated with -,
-hospolipase C performs a catalytic mechanism! generating inositol triphosphate 08-91
and diacylglycerol 05&'1$
8n this cascade! 5&' remains on the cell membane and activates the signal cascade by
activating protein #inase C 0-+C1$ -+C in turn activates other cytosolic proteins by
phosphorylating them$ The effect of -+C could be reversed by phosphodiesterases
0-5E1$
8-9 enters the cytoplasm and activates 8-9 receptors on the smooth endoplasmic reticulum 0E"1!
which opens calcium channels on the smooth E"! allowing mobili4ation of calcium ions through
specific Ca(E channels into the cytosol$ Calcium participates in the cascade by activating other
proteins$
-8-( can be further phosphorylated by -89 #inase 0-89+1 to yield phosp hatidylinositol 09!A$=1-
triphosphate 0-8-91 7nce formed ! the phosphorylatedF head groupF of -8-9 serves to attract
proteins carrying -6 domains! w hich thereby become tetheredto the inner surface of the plasma
membrane$ -lec#strin homology 0-61 -6 domain-containing protein is the serine-threonine
#inase named &#t! also #nown as protein #inase : 0-+:1$ This association of &#tl-+: with the
plasma membrane 0together with phosphorylations of &#t<-+: that soon follow1 results in the
functional activation of &#t<-+: as a #inase$ 7nce activated by these phosphorylations! &#tl-+:
proceeds to phosphorylate a series of protein substrates that have m@ltiple effects on the cell$
Three major biological effects that &#tl-+: has on cells are
0)1 aiding in cell survival by reducing the possibility that the cell apoptotic suicide
program will become activatedG
0(1 stimulating cell proliferationG and
091 stimulating cell growth!$
phosphatases! -TEN! removes the 9H phosphate group from -8-9 in cancer cells- hyperactivity of
-89+ or inactivity of -TEN$ -89+ is hyperactivated in almost one-third of human colon
carcinomas$ -TEN activity is lost because of chromosomal gene mutations or 5N& methylation
events that suppress expression of the -TEN gene$ &ltogether! -TEN activity is lost in 9I to AI*
of all human cancers$
Rho protein! &mong the -6-bearing cellular proteins are a group of guanine nucleotide
exchange factors 0'E2s1 that function analogously to os! being responsible for activating
various small 'T-ases that are distant relatives of the "as proteins$ These other 'T-ases belong
to the "ho family of signaling proteins! which includes "ho proper and its two cousins! "ac and
CdcA($ they participate in reconfiguring the structure of the cytos#eleton and the attachments that
the cell ma#es with its physical surroundings$ 8n so doing! these "ho-li#e proteins control cell
shape! motility! and! in the case of cancer cells! invasiveness$ 2or example! CdcA( is involved in
reorgani4ing the actin cytos#eleton of the cell as well as controlling filopodia! small! fingerli#e
extensions from the plasma membrane that the cell uses to explore its environmentG while "ac is
involved in the formation of lamellipodia! broad ruffles extending from the plasma membrane
that are found at the leading edges of motile cells$
A third Ras-reg"lated pathway in#ol#es a pair of Ras-like proteins termed Ral-A and Ral-.
2unctional activation of these "al proteins involves replacement of bound '5- with
'T-$
Communication between "as and "al is mediated by "al 'uanine nucleotide exchange
factor which can stimulate a "al protein to shed its '5- and bind 'T-$
"as binds to "al-'E2 and cause a conformational shift in "al-'E2 that activates their
intrinsic 'uanine nucleotide exchange factor 0'E21 activity
The "al pathway can inactivate Jtwo of the "ho proteins cdcA( and "ac$
"ac can also emit mitogenic signals and by stimulating the production of "7
!antagonise the actions of several "ho proteins$
"al proteins play #ey roles in the motility that enable cancer cells to invade and metasi4e$
"ac can also emit mitogenic signals and by stimulating the production of "7
!antagonise the actions of several "ho proteins$
$
,ultiple '&-s! and 'E2s have been shown to undergo overexpression! downregulation!
or mutation in different types of cancer $
Tumor modification of rho activity can suppress apoptosis and therefore contribute to
artificial cell longevity
&fter natural apoptosis is suppressed! abnormal tumor growth can be observed through
the loss of polarity in which rho proteins play an integral role$
The growing mass can invade across its normal boundaries through the alteration of
adhesion proteins potentially caused by rho proteins$
2inally! after inhibition of apoptosis! cell polarity and adhesion molecules! the cancerous
mass is free to metastasi4e and spread to other regions of the body$
$nt-%- catenin pathway
8n addition to transducing mitogenic signals! this pathway enables cells to remain in a relatively
undifferentiated state-an important attribute of certain types of cancer cells$ Bnt factors! acting
through 2ri44led receptors! suppress the activity of glycogen synthase #inase-B 0'+-9?1$
Cadherins are transmembrane that form attachments in the extracellular space and become lin#ed
via intermediary proteins to the actin cytos#eleton$ K-catenin along with L-catenin and p-)(I
form the mechanical lin#age between cadherins and actin cytos#eleton 8n the absence ofBnts!
'+9? phosphorylates several #ey substrate proteins! which are thereby tagged for destruction$
The most important of these substrates is ?-catenin! normally a cytoplasmic protein that exists in
three states$ 8t may be bound to the cytoplasmic domain of cell-cell adhesion receptors! such as E-
cadherin 02igure 3$(3&1$ &lternatively! in a fully unrelated role! it exists in a soluble pool in the
cytosol! where it turns over very rapidly! having a lifetime of less than (I minutes$ &nd
finally! ?-catenin operates in the nucleus as an important component of a transcription
factor$ Bhen Bnt signaling is activated! however! '+-9? firing is bloc#ed and ?catenin
is saved from rapid destructionG its half-life increases from less than (I minutes to )-( hours! and
its steady-state concentrations increase proportionately$ ,any of the accumulated ?-catenin
molecules then move into the nucleus to activate transcription by binding to Tcf<Lef proteins 0see
2igure 3$(3:1$ The resulting multi-subunit transcription factor complexes proceed to activate
expression of a number of important target genes! including those encoding
critical proteins involved in cell growth and proliferation! such as cyclin 5l and
,yc!
Bnts is increased four to ten fold above normal in breast cancer
Clear evidence of nuclear translocation of M%catenin in (I* of prostrate carcinomas
8n =-N* prostrate carcinomas mutations of M%catenin occurs0liver!colon carcinoma!and
melanoma$
&ll colon carcinomas M%catenin degradn is defective-due to defects in apc
&-protein-co"pled receptors dri#e normal and neoplastic proliferation
'-protein %coupled receptors 0'-C"s1 are transmembrane proteins that weave their way
bac# and forth through the plasma membrane seven times$
@pon binding their extracellular ligands! each of these serpentine receptors activates one
or more types of cytoplasmic heterodimeric ' protein! so named because of its three
distinct subunits0'- L!'-M and 'O1!the first of which binds either '5- or 'T-$
The activated state of ' L is achieved when it binds 'T-$
7nce stimulated by a '-protein-coupled receptor 0'-C"1! the '- L subunit of a
heterotrimeric ' protein dissociates from its two partners!' M and '- O and proceeds to
activate a number of distinct cytoplasmic en4ymes$
ome of these are adenyl cyclase which converts &T- into c-&,- and phospholipase C-M
which cleaves -8--( to yield 5&' and 8-9$
The latter are potent second messengers that can function to stimulate cell proliferation$
'-protein %coupled receptors 0'-C"s1 are transmembrane proteins that weave their way
bac# and forth through the plasma membrane seven times$
@pon binding their extracellular ligands! each of these serpentine receptors activates one
or more types of cytoplasmic heterodimeric ' protein! so named because of its three
distinct subunits0'- L!'-M and 'O1!the first of which binds either '5- or 'T-$
$
$The rc +inase !the first cellular oncoprotein can be activated by certain 'T--bound '-L
subunits$
&t the same time!complexes of the other two subunits of the heterotrimeric ' protein-'-M
and '-O have been found to stimulate yet other important mitogenic signaling proteins!
such as one form of phosphatidylinositol 9-#inase 0-89+1$
The ability of '-C"s to activate mitogenic pathways suggests that deregulation of
signaling by these receptors and associated ' proteins may well contribute to cell
transformation and tumorigenesis$
Cancer cells of small-cell lung carcinomas! a common tumor of cigarette smo#ers!release
a number of distinct peptide factors! some with neuropeptide li#e properties
8n CLC!s the tumor cells may simulataneously secrete bombesin also termed as
'"-!brady#inin!gastrin! neurotensin and vasopressin$
&t the same time these cells display the '-C"s that recognise and bind these released
factors resulting in the establishment of autocrine signaling loops$
CLC cells incubated with antibody that binds and neutralises a secreted autocrine
growth factor led to cessation of growth
8n thyroid adenomas and carcinomas T6" and '-C" have point mutations leading to
ligand independent firing
'o"r more . they control the intracellular locali4ation of Fdual-addressF proteins that
operate much li#e ?-catenin. these proteins normally reside in the cytoplasm! and when activated
by certain signals! they are dispatched to the nucleus! where they function as components of
specific transcription factors to drive gene expression$
("clear factor- k)eta . Components of this cascade are rarely found in mutant forms in
human cancer$
This pathway is often constitutively activated$
8n breast cancer this pathway is highly active
,alignancies of lymphocyte lineages
,any other pre-malignant growths
The "EL gene which encodes one of the subunits of N2-+: seems is amplified in about
one-fourth of diffuse large :-cell lymphomas resulting in a A-to 9= fold increase in gene
expression$
(otch is a transmembrane proteinG four different varieties of Notch 0products of four different
genes1 are expressed by mammalian cells$ &s mentioned in the last chapter! after Notch! acting as
a cell surface receptor! binds a ligand 0NotchL1! it undergoes two proteolytic cleavages! one in its
ectodomain! the other P$vithin its transmembrane domain$ The latter cleavage liberates a largely
cytoplasmic protein fragment from its tethering to the plasma membrane$ This fragment of Notch
then migrates to the nucleus of targeted cells! where it functions! together with partner proteins! as
a transcription factor$ 7verexpression in cervical carcinoma!colon carcinoma!squamous
carcinoma of lung$ This overexpression is often accompained by nuclear localisation of the
cytoplasmic cleavage fragment of Notch! indicating active signaling through this pathway$
/agged and delta mutations cervical and prostrate cancer 8n )I* acute Lymphocytic Leu#emia
have constitutively active forms of notch$ This is because of the genetic deletions of the portions
of the N7TC6-) gene encoding the extracellular domain of the Notch-) protein$
the Patched receptor by its ligand! 6edgehog! causes -atched to release the moothened protein
from inhibitionG moothened is then able to emit downstream signals$ Through a poorly
understood series of changes! the activated moothened protein alters the fate of cytoplasmic 'li
protein$ Normally! in the absence of intervention by moothened! the 'li precursor protein is
cleaved into two fragments! one of which moves into the nucleus! where it functions as a
transcriptional repressor$ 6owever! when moothened is activated! it protects the 'li precursor
from being cleaved$ The resulting intact 'li protein migrates to the nucleus! where it
serves as an activator of transcription$
6ighly expressed in 'lioblastomas
,edulloblastomas
:asal cell carcinomas
#in tumours
Tumors of cerebellum
:reast and esophageal carcinomas
T&' * . T'2-? acts to arrest the growth of many cell types! and later in cancer progression!
when it contributes! paradoxically! to the phenotype of tumor invasiveness$ activation
of this pathway leads to dispatch of mad transcription factors to the nucleus! where they can
collaborate with other nuclear transcription factors to activate the expression of a large
constituency of genes 02igure 3$(Q51$ 8n the absence of critical mads! epithelial cancer cells can
escape the growthinhibitory actions ofT'2-? and thriv