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ONCOGENIC PI3K DEREGULATES

TRANSCRIPTION AND
TRANSLATION
Andreas G. Bader, Sohye Kang, Li Zhao and Peter K. Vogt
Abstract | There have long been indications of a role for PI3K (phosphatidylinositol 3-kinase) in
cancer pathogenesis. Experimental data document a requirement for deregulation of both
transcription and translation in PI3K-mediated oncogenic transformation. The recent
discoveries of cancer-specific mutations in PIK3CA, the gene that encodes the catalytic
subunit p110 of PI3K, have heightened the interest in the oncogenic potential of this lipid
kinase and have made p110 an ideal drug target.

SH2 DOMAIN
A protein domain, homologous
to an equivalent domain in the
SRC protein, that facilitates
proteinprotein interactions by
binding to tyrosine-
phosphorylated protein
sequences.
PLECKSTRIN HOMOLOGY
DOMAIN
A protein domain that was
originally identified in the
protein pleckstrin that forms a
phospholipid-binding pocket
and in particular binds to PIP3.
Department of Molecular
and Experimental
Medicine, The Scripps
Research Institute,
10550 North Torrey Pines
Road, La Jolla,
California 92037, USA.
Correspondence to P.K.V.
e-mail: pkvogt@scripps.edu
doi:10.1038/nrc1753
Phosphatidylinositol 3-kinases (PI3Ks) phospho-
rylate phosphatidylinositol and its phosphorylated
derivatives at the 3 position of the inositol ring,
generating second messengers that control cellular
activities and properties including proliferation, sur-
vival, motility and morphology mutations in PI3K
function disrupt these processes110. PI3K proteins
form a family that is divided into three classes that
differ in structure, substrate preference, tissue distri-
bution, mechanism of activation and, ultimately, in
function1115 (FIG. 1). In regulating proliferation and
in tumorigenesis, the most important PI3K proteins
are those that belong to class IA the catalytic
subunit p110 and its associated regulatory subunit
p85. In quiescent cells, p85 binds to p110, stabiliz-
ing p110 and inactivating its kinase activity. Upon
growth factor stimulation, the SH2 DOMAINS (Rous-
sarcoma-oncogene homology-2 domains) of p85 bind
to phosphorylated tyrosine in YxxM motifs in recep-
tor tyrosine kinases or their substrate adaptor pro-
teins. This binding relieves the inhibition of p110
and mediates recruitment of the catalytic subunit
to the plasma membrane16. PI3K activity is further
augmented by the interaction between p110 and the
GTP-bound form of the RAS oncoprotein17,18.
The high-affinity substrate of p110 is phos-
phatidylinositol 4,5-bisphosphate (PIP 2 ). This is
phosphorylated to become phosphatidylinositol
3,4,5-trisphosphate (PIP 3 ), which then recruits
proteins that contain a PLECKSTRIN HOMOLOGY DOMAIN
to cellular membranes 19 . Among these are the
serine-threonine kinase AKT, as well as its activat-
ing kinases PDK1 (3-phosphoinositide-dependent
kinase 1) and possibly PDK2. The identity of the lat-
ter has remained controversial. Potential candidates
for PDK2 include PDK1, AKT, integrin-linked kinase
(ILK), DNA-dependent protein kinase (DNA-PK)
and the RictorTOR (target of rapamycin) com-
plex2024. Recruitment of these kinases by PIP3 results
in the activation of AKT by phosphorylation at T308
and S473. Activated AKT is the predominant and
essential mediator for the regulation of both growth
and proliferation by PI3K. PIP3 is also a substrate
of the phosphatase PTEN (phosphatase and tensin
homologue deleted on chromosome 10), which
dephosphorylates PIP 3 to generate PIP 2. PTEN is
a negative regulator of PI3K signalling and func-
tions as a tumour suppressor25,26. Genetic inactiva-
tion of PTEN occurs in numerous human tumours
and results in a constitutive upregulation of PI3K
signals2730.
Wild-type p110 is not oncogenic, but its
growth-regulatory functions confer a potential for
oncogenicity that can be activated by simple genetic
changes. These include retroviral transduction, point
mutations, gene amplification, overexpression and

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Summary constitutive activation of PI3KAKT signalling36,3840.

Phosphatidylinositol 3-kinases (PI3Ks) are lipid kinases that generate second
messengers that govern cellular activities and properties including proliferation,
survival, motility and morphology.
PIK3CA, the gene that encodes the catalytic subunit p110 of PI3K, is frequently
mutated in human solid tumours.
Cancer-specific mutations are clustered in the helical and the kinase domains of
p110. The amino-acid residues E542, E545 and H1047 are prominent mutational
hot spots.
The mutations in the p110 hot spots induce a gain of enzymatic function and
confer oncogenic activity both in vitro and in vivo.
The oncogenicity of PI3K is the result of interactions with transcriptional and
translational controls.
the acquisition of sequences that localize the pro-
tein to cellular membranes. The avian PIK3CA gene
and the mouse Akt gene have both been identified
as oncogenes in retroviruses3133. Oncogenic forms
of p110 and AKT are constitutively present in the
cell membrane where they have high levels of kinase
activity34,35.
Mutations in the regulatory subunit of PI3K, p85, have
been identified in transformed cell lines and in human
tumour samples3640. The mutant forms of p85 induce
Expression of mutant p85 in mice induces a lympho-
proliferative disorder and solid tumours36,37.
The gain of function in PI3KAKT signalling,
which is characteristic of experimental transformation
in cell culture and of many human tumours, affects the
synthetic activities of the cell at the transcriptional and
translational levels (FIG. 2).
PI3K and transcription
PI3KAKT signals control several growth-regulatory
transcription factors. Two prominent examples are the
forkhead box (FOXO) proteins and nuclear factor
(NFB). The FOXO transcription factors, which are
a subclass of the much larger FOX protein family,
encompass FOXO1 (FKHR), FOXO3A (FKHRL1),
FOXO4 (AFX) and FOXO6 REF. 41. FOX proteins
bind to DNA as monomers with a highly conserved
domain that is referred to as the forkhead or winged
helix domain that also serves as the defining struc-
tural feature of the protein family. The FOXO proteins
control the pace of the cell cycle, as well as apoptosis,
DNA repair and protection from oxidative damage42,43.
Their activities are regulated by post-translational
modifications that include phosphorylation by sev-
eral kinases, acetylation and ubiquitylation. These

Subunits

Class Structural features of catalytic subunits Catalytic Adaptor Regulation Lipid substrate
specificity

I Adaptor- p110,, p85 Receptor PIP2

binding C2 Catalytic p55 tyrosine PIP
domain domain domain p50 kinases and RAS PI
p85 (p110 is also
A p55 regulated by
Helical G-protein-coupled
RBD receptors)
domain

p110 p101 G-protein-coupled PIP2

B p84 receptors and PIP
RAS PI

Coiled-coil PX
domain domain
II PI3K-C2,, ? Receptor tyrosine PIP2
kinases? PI
Proline-rich C2 G-protein-coupled
domain domain receptors?

III Vps34p p150 Constitutive? PI

analogues

Figure 1 | PI3Ks constitute a family of enzymes that is divided into three classes. The figure summarizes the domain
organization and defining criteria of the three phosphatidylinositol 3-kinase (PI3K) classes. The catalytic subunit of class I
enzymes (p110, , and ) contains an adaptor-binding domain, a RAS-binding domain (RBD), a C2 (protein-kinase-C
homology-2) domain, a helical domain and a catalytic domain. Class IA enzymes are constitutively associated with an adaptor
subunit to form a heterodimeric complex. There are three genes that encode at least five different adaptor subunits, all of which
can bind to phosphorylated tyrosine in YxxM motifs through their SH2 (Rous-sarcoma-oncoprotein homology-2) domains.
The only member of class IB is p110, which associates with the p101 regulatory subunit or the recently identified p84 adaptor
subunit. Class I enzymes are regulated by receptor tyrosine kinases and RAS, and have lipid substrates that include
phosphatidylinositol-bisphosphates (PIP2), phosphatidylinositol phosphate (PIP) and phosphatidylinositol (PI). Class II enzymes
are comprised of a PI3K-C2, and domains. They lack an adaptor-binding site but possess carboxy-terminal phox (PX) and
C2 domains that could mediate binding to phosphoinositides that have been phosphorylated at position D3 and other
membrane lipids. Class II enzyme substrates include PIP2, PIP and PI. Although their adaptor subunits have not been identified,
class II enzymes are believed to be regulated by receptor tyrosine kinases and G-protein coupled receptors. Class III enzymes
comprise analogues of the yeast Vps34p PI3K subunit and a p150 adaptor subunit, and accept only PI as a substrate. They are
believed to be constitutively active, and all isoforms of class I PI3K also possess an intrinsic protein kinase activity.

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modifications determine the cellular localization
and, ultimately, the fate and functions of FOXO pro-
teins nuclear FOXO proteins act as transcriptional
regulators whereas cytoplasmic FOXO proteins are
transcriptionally silent and are subject to proteasomal
degradation.
PIP2 PI3K PIP3 PIP3 PIP3
AKT PDK
PTEN
TSC
GSK3
MDM2
RHEB
IKK
TOR
NFB
S6K
p53
4EBP MIZ1
S6 FOXO c-MYC
HIF1 AP1
4E -catenin
Translation Transcription
Figure 2 | PI3KAKT signalling affects translation and
transcription. The lipid kinase phosphatidylinositol 3-kinase
(PI3K) phosphorylates phosphatidylinositol-bisphosphates
(PIP2), generating phosphatidylinositol-trisphosphates (PIP3),
which recruit the serine-threonine kinase AKT and
3-phosphoinositide-dependent kinase (PDK) to the plasma
membrane. Phosphorylation by PDK activates AKT. The
signal branches at the level of AKT, regulating downstream
proteins that control translation and transcription. During
translation, AKT signalling stimulates protein synthesis by
activating the ribosomal protein S6 and the translational
initiation factor 4E. Stimulation of these translational
regulators is indirect and involves the signalling molecules
TSC (tuberous sclerosis complex), RHEB (RAS homologue
enriched in brain) and TOR (target of rapamycin). TSC is a
heterodimeric complex consisting of TSC1 (also known as
hamartin) and TSC2 (also known as tuberin) and functions
as a GTPase-activating protein for the small GTPase
RHEB155159. TSC2 is directly phosphorylated and
inactivated by AKT, which leads to increased RHEB
activity155,160. In the GTP-bound state, RHEB activates TOR,
presumably by direct interaction161. TOR then mediates
phosphorylation of S6K and 4EBP (4E-binding protein),
thereby stimulating the former and inactivating the latter,
leading to increased S6 and 4E activity. During transcription,
AKT affects numerous transcription factors, the control of
which is either direct or indirect. Direct targets are the
forkhead box proteins, FOXO, and the cell cycle inhibitor,
MIZ1, both of which are inhibited upon AKT-mediated
phosphorylation. AKT-dependent regulation of p53, nuclear
factor B (NFB), c-MYC, activator protein 1 (AP1) and
-catenin is indirect and also independent of TOR. However,
TOR does regulate the transcriptional activity of the hypoxia-
inducible factor HIF1162,163. GSK3, glycogen synthase
kinase 3; IKK, IB kinase; MDM2, murine double minute;
PDK, 3-phosphoinositide-dependent kinase; PTEN,
phosphatase and tensin homologue deleted on
chromosome 10.
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Nuclear FOXO proteins activate several distinct
transcriptional programmes. One of these leads
to G1 arrest through increased expression of the
cyclin-dependent kinase inhibitors p27kip1 and p21cip1,
and reduced expression of cyclin D1 and cyclin D2
REFS 4447. AKT terminates these activities by initiat-
ing nuclear export of FOXO proteins. It phosphorylates
FOXO at three conserved residues4852, thereby mask-
ing the nuclear localization signal, interfering with
DNA-binding, disrupting the association with the
co-activator p300CBP (CREB-binding protein) and
mediating the interaction with 14-3-3 proteins and the
exportin CRM1 REFS 49,5358. The result is inhibition
of FOXO nuclear functions and removal of FOXO
from the nucleus.
FOXO that has been exported from the nucleus
as a result of AKT-dependent phosphorylation is
rapidly degraded5962. The phosphorylation by AKT
not only triggers nuclear export of FOXO but it also
creates a binding site for the ubiquitin ligase S-phase
kinase-associated protein 2 (SKP2, a suppressor of
a cyclin-dependent-kinase-inhibitor proteolysis
defect), which then marks FOXO for proteasomal
degradation60. This radical elimination of transcrip-
tionally active FOXO is a key feature of cells that
have been transformed by oncogenic PI3K or AKT
in these cells, levels of FOXO are dramatically
reduced and the growth-regulatory activity of FOXO
is suspended59. FOXO activity also influences other
signalling pathways in a complex with SMAD,
FOXO regulates p21cip1 gene expression during TGF
signalling, and FOXO3a is inactivated by the NFB
pathway45, 63.
Whereas FOXO is inactivated by AKT signalling, the
transcription factor NFB is activated. In human cancer
cells, NFB positively regulates cell survival, prolifera-
tion, chemoresistance, angiogenesis, cellular invasion
and oncogenesis6466. NFB is negatively regulated by
the inhibitor IB, which anchors NFB in the cyto-
plasm. Positive regulation of NFB occurs through IB
kinases (IKKs), which phosphorylate IB and mark it
for proteasomal degradation. Released from IB, NFB
travels into the nucleus and forms a functional tran-
scriptional unit with its dimerization partner, REL-A,
which is also known as p65 REFS 64,65.
AKT stimulates the transcriptional activity of NFB
by three distinct modes, all of which require IKK func-
tion. First, AKT interacts with the IKK complex, phos-
phorylates the subunit and increases IKK activity67,68.
Next, AKT indirectly stimulates IKK activity through
the mitogen-activated protein kinase kinase kinase
COT69. Finally, AKT targets the transactivation domain
of REL-A, increasing NFB activity7072.
In specific cellular settings, the transcriptional activ-
ity of NFB is required for oncogenic transformation by
PI3KAKT signalling. For example, disruption of the
nuclear activity of NFB with peptide antagonists of
its nuclear localization signal, or with non-degradable
forms of IB, inhibits oncogenic transformation and
cell proliferation, and induces apoptosis in pancreatic
cancer cells73,74. NFB activation by PI3KAKT can also
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Table 1 | Aberrant PI3K-signalling in human cancers
Component Type of alteration References
p110 Gain of function by amplification, 110,143,144,164166
overexpression or somatic mutation
p85 Gain of function by mutation 3640
PTEN Loss of function by mutation or 2729,158,167
transcriptional downregulation
AKT Gain of function by amplification, 144,167170
overexpression or increased
phosphorylation
AKT, serine-threonine kinase that is a homologue of thymoma viral oncoprotein; p110, catalytic
subunit of phosphotidylinositol 3-kinase (PI3K); p85, regulatory subunit of PI3K; PTEN,
phosphatase and tensin homologue deleted on chromosome 10.
result in a positive auto-feedback loop NFB signal-
ling indirectly represses transcription of the PTEN gene
and therefore amplifies the PI3K signal75,76.
Other transcriptional regulators whose activities are
affected by PI3KAKT signalling include MIZ1 (Myc-
interacting zinc-finger protein 1), p53, AP1 (activator
protein 1), c-MYC, -catenin and HIF1 (hypoxia
inducible factor 1). The exact roles of these proteins
during PI3K-mediated oncogenesis are currently
unknown, but they have all been linked to oncogenic
transformation. AKT phosphorylates and thereby inac-
tivates the cell-cycle inhibitor MIZ1 REF. 77, and also
suppresses p53 activity by a mechanism that involves
MDM2 (murine double minute)78,79. By contrast, the
activities of AP1, c-MYC and -catenin are increased
by AKT. These targets are negatively controlled by
GSK3 (glycogen synthase kinase 3), which is inacti-
vated by AKT-mediated phosphorylation8087. In cancer
cell lines that upregulate PI3K signalling, levels of HIF1
are increased, and the gene targets of this transcrip-
tional regulator (for example, vascular endothelial
growth factor) are activated88.
PI3K and protein synthesis
The key evidence that links protein synthesis to
PI3K-mediated growth signals comes from studies
MACROLIDE
with rapamycin and its target, TOR. Rapamycin is a
A group of antibiotics that are
produced by various strains of MACROLIDE antibiotic that interacts with a small cellu-
Streptomyces and have a lar protein FKBP12 (FK506-binding protein 12)89,90.
complex macrocyclic structure. The rapamycinFKBP12 complex binds to the kinase
TOR, interfering with the TOR-Raptor (regulatory
POL I AND POL IIIDEPENDENT
associated protein of TOR) connection and block-
TRANSCRIPTION
Class I RNA polymerases
ing the phosphorylation of TORRaptor targets9196.
synthesize all ribosomal RNAs TOR has multiple and diverse functions, including the
except the 5S ribosomal RNA; control of protein synthesis97. It stimulates POL I AND
class III RNA polymerases POL IIIDEPENDENT TRANSCRIPTION, thereby regulating the
transcribe transfer RNA genes
abundance of the components that make up the pro-
and the gene that encodes the
5S ribosomal RNA. tein-synthesizing machinery of the cell98102.
The activities of Pol I and of Pol III have long-term
mRNA CAP STRUCTURE effects, but TOR also regulates protein synthesis on
The 5 end of virtually all a shorter timescale by inducing the phosphorylation
mRNAs is protected by a cap of S6K (p70 S6 kinase) and of the 4EBP (translation
with the chemical structure initiation factor 4E-binding protein) family of pro-
m7GpppN (where m7G
represents 7-methylguanylate, p
teins97,103. S6K is a positive regulator of protein syn-
represents a phosphate group thesis and cell size104107. Its mechanism of action is not
and N represents any base). fully understood; recent experiments have challenged
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previously proposed models in which S6K controls
the translation of mRNAs that contain a 5-terminal
oligopyrimidine tract adjacent to the mRNA CAP STRUC
108,109
TURE . The 4EBP proteins are negative regulators of
protein synthesis. They interact with the cap-binding
protein 4E and thereby prevent the formation of the
4F translational initiation complex, which depends on
the availability of 4E110. The phosphorylation of 4EBP
by TOR breaks the attachment to 4E, allowing 4E to
build a functional initiation complex and recruit the
scaffolding protein 4G and the RNA helicase 4A111113.
TOR signals to its targets, S6K and 4EBP, through
interaction with a binding partner, Raptor114,115. The
function of Raptor in this process depends on a spe-
cific motif in the target proteins, named the TOS (TOR
signalling) motif116118 (FIG. 2).
The oncogenic activities of PI3K and of AKT are
exquisitely sensitive to rapamycin. Low concentrations
of the drug completely prevent transformation that is
induced by oncogenic versions of PI3K and of AKT
in cell culture119. However, rapamycin does not inhibit
transformation that is induced by other oncoproteins,
although some, such as SRC, are sensitive to rapamycin
in combination with inhibitors of MAP kinase signal-
ling119,120. Sensitivity of tumours to rapamycin is also
seen in animal model systems121,122. PTEN-deficient
mice develop spontaneous cancers that show consti-
tutive gain of function in PI3KAKT signalling. The
tumours respond to treatment with the rapamycin
derivative CCI-779, which shows that TOR activity is
essential for PI3K-dependent oncogenesis. In PI3K-
transformed or AKT-transformed cells, S6K and 4EBP
are constitutively phosphorylated, indicating activation
of the former and inactivation of the latter119. This con-
stitutive phosphorylation is sensitive to rapamycin, and
is therefore TOR-dependent119.
The correlation between TOR-dependent inter-
vention in protein synthesis and PI3K-induced
oncogenic transformation is complemented by data
on another regulator of protein synthesis, the mRNA-
binding protein YB-1 (Y box-binding protein 1)123126.
YB-1 is transcriptionally downregulated in PI3K-
transformed and AKT-transformed cells127. Vector-
mediated expression of YB-1 in normal cells induces
specific resistance to transformation by PI3K or AKT
but does not affect transforming activities of other
oncoproteins127. YB-1 acts downstream of TOR, as the
phosphorylation levels of the TOR targets S6K and
4EBP are not changed in YB-1-overexpressing cells127.
A deletion analysis of YB-1 shows that interference
with PI3K-induced and AKT-induced transforma-
tion is correlated with the ability of YB-1 to inhibit
cap-dependent protein synthesis128. All interfering
YB-1 mutants also bind to mRNA, including the cap
structure, and are localized in the cytoplasm128. The
data on YB-1 provide an independent line of evidence
for an essential role of protein synthesis in PI3K-
induced and AKT-induced transformation. It is likely
that this role does not extend to protein synthesis in
general but is restricted to the efficient translation of
specific mRNAs.
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Microarray analysis of polysomal RNA samples have 4E, which recruits an RNA helicase that can unwind
shown a differential effect of PI3K, AKT and rapamy- 5-mRNA structures to facilitate ribosome scanning132.
cin treatment on the expression of specific mRNAs129 Numerous mRNAs that code for growth factors, onco-
131
. These included mRNAs with lower translational proteins and cell cycle regulators have highly struc-
fitness, presumably due to highly structured, com- tured 5-untranslated regions, making their efficient
plex 5-untranslated regions. Efficient translation of translation dependent on the abundance of 4E132.
such mRNAs is dependent on high levels of active We conclude that deregulation of protein synthesis
is necessary for PI3K- induced and AKT-induced
p110 Frequency of mutation (%) transformation, but it might not be sufficient, as there
are no reports that show oncogenic transformation by
Mutated residue Colon Breast Others Total TOR or the TOR activator, RHEB (RAS homologue
enriched in brain).
R38 1.1 0.13
The effects of AKT on transcriptional regulators are
Q60 0.2 0.04
p85 R88 0.4 0.2 0.1 0.13
also essential features of the transformation process
P104 0.4 0.04 transcriptional and translational deregulation are both
G106 0.4 0.04 necessary and complement each other. In addition to
R108 0.4 0.04 deregulating transcription and protein synthesis, onco-
E110 0.2 0.04 genic PI3K also affects cell replication and survival by
K111 0.2 0.04 post-translational modification of proteins. Examples
RBD G118 0.4 0.04 of these effects that are outside the scope of this review
G122 0.4 0.04 are the AKT-induced phosphorylation and inactiva-
P124 0.4 0.04
tion of the cyclin-dependent kinase inhibitors p21cip1
N345 0.4 0.3 0.13 and p27kip1, as well as of the pro-apoptotic proteins
D350 0.1 0.04 BAD (BCL2-anatagonist of cell death) and caspase 9
C378 0.1 0.04
C2 REFS 133140.
S405 0.2 0.04
E418 0.2 0.04
C420 0.8 1.2 0.39 PI3K as a drug target
E453 0.4 0.2 0.09 Genetic aberrations that lead to a gain in PI3K
signalling are commonly observed in human can-
P539 0.4 0.3 0.13 cers TABLE 1. The recent sequencing of cancer cell
E542 3.8 3.6 0.5 1.63 genomes has revealed point mutations in PIK3CA110.
E545 9.8 6.2 3.3 4.76 These mutations occur at significant frequencies
Helical Q546 2.6 0.2 0.2 0.47 in several types of solid tumours (FIG. 3). They are
observed in colon as well as in breast and hepato-
Q661 0.4 0.04
cellular carcinomas. The mutations in PIK3CA in
H701 0.3 0.1 0.13 human tumours are somatic, cancer-specific and
K733 0.4 0.04 heterozygous. They are mostly missense mutations;
no truncating or nonsense mutations have been
C901 0.4 0.1 0.09 identified, but a few cases of in-frame deletions
F909 0.4 0.2 0.09
and insertions have been detected1,3,5. Rare cases of
S1008 0.4 0.04
double mutations, in which two amino-acid resi-
P1011 0.1 0.04
Y1021 0.2 0.13 dues are altered, have also been reported5,8. In breast
Kinase carcinoma, mutations in PIK3CA and loss of PTEN
T1025 0.8 0.2 0.1 0.17
E1035 0.1 0.04 function are almost always mutually exclusive 8 .
M1043 0.8 0.2 0.1 0.21 However, mutations in PIK3CA are often correlated
N1044 0.2 0.04 with overexpression of ERBB2 (also known as HER2/
A1046 0.2 0.04 NEU), indicating that gains of function in these two
H1047 7.1 14.8 1.5 5.48
signalling components could have a synergistic effect
G1049 0.2 0.1 0.09
H1065 0.1 0.04
in counteracting PTEN8.
The most important attribute of the cancer-
Total 32.3 29.3 6.5 15.1 specific mutations in PIK3CA is a non-random
Figure 3 | Point mutations in PIK3CA observed in human tumours. Analyses of the phos-
distribution along the coding sequence (FIG. 3). This
phatidylinositol 3-kinase PIK3CA gene in human tumour samples have identified somatic point non-randomness is indicative of selection for muta-
mutations that affect a total of 38 residues110. Mutations other than point mutations have not tions that confer a proliferative advantage, which
been included in the figure. The mutations localize to various domains of the p110 primary act as dominant oncoproteins. The majority of the
structure as indicated. Hot-spot mutations that occur at high frequency are confined to mutations map to three sites E542 and E545 in
residues E542, E545 and H1047 and are highlighted in orange. Overall frequencies of mutation the helical domain and H1047 in the kinase domain of
have been determined for cancers of the colon (266 samples)1,3, breast (580 samples)13,5,6,8
p110 (FIG. 3). E542 and E545 are commonly changed
and others, which include liver (73 samples)5, brain (382 samples)1,4,10, stomach (291
samples)1,5,7, lung (253 samples)1,5 and ovary (489 samples)3,6,9, with a total of 2,334 tumour to lysine, whereas H1047 is frequently substituted
samples. Protein domains have been determined using NCBI tools. C2, protein-kinase-C with arginine. At all three sites, the mutations cause
homology-2 domain; p85, p85-binding domain; RBD, RAS-binding domain. a gain of enzymatic function1,141. The three mutant

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proteins also induce oncogenic transformation when
expressed in primary chicken-embryo fibroblasts and
in NIH 3T3 cells141,142. AKT signalling is constitutively
activated in the mutant-transformed cells, as indicated
by high levels of phosphorylated AKT and of its down-
stream targets S6K and 4EBP141,142. The oncogenic
transformation that is induced by the three mutant
p110 proteins is inhibited by rapamycin, indicat-
ing that TOR and TOR-dependent protein synthesis
are essential components of the transformation
process141.
Mutations in p110 and other mechanisms of acti-
vating PI3K signalling appear to be mutually exclusive.
In ovarian cancer, mutations in p110 are confined
almost exclusively to tumours that do not show ampli-
fication of the PIK3CA gene3. Since amplification of
PIK3CA is common in ovarian cancers, mutations
in p110 are rare in these tumours3, 9, 143. Likewise, in
gastric tumours, the incidence of PIK3CA amplifica-
tion far exceeds that of mutations in PIK3CA5,7,144. By
contrast, tumours that lack PIK3CA amplification,
such as in breast cancer or colorectal cancer, more
frequently harbour mutations in p1103, 6.
The mechanisms by which the mutations in p110
induce a gain of function remain to be determined. In
the absence of structural data for p110, it is neverthe-
less possible to model the catalytic and helical domains
of the molecule using the published crystal structure of
the homologous p110 isoform145 (FIG. 1). In this model,
the two most prevalent mutations in the helical domain,
E542K and E545K, map to the surface of the protein6,146.
Both mutations cause an electrostatic charge reversal
from negative to positive that could change the abil-
ity of the protein to interact with regulatory proteins.
Although deletion mapping has defined separate bind-
ing regions for the regulatory subunit p85 and for RAS
that lie in the amino-terminus of p110, the data do
not rule out an extended interaction of p85 or RAS with
this helical domain. The p85 subunit can function as a
negative regulator of p110, and reduction of p85 bind-
ing by the E542K or E545K mutations could increase
p110 enzymatic activity147. There is also the possibil-
ity that these mutations disrupt the ability of p110
to interact with a still unknown regulatory protein.
Another possible explanation for the gain of function
that is induced by the mutations in the helical domain
is that these cause a change in enzyme conformation.
These mutations could affect the C2 (protein-kinase-C
homology-2) domain and thereby increase the affin-
ity of that domain for lipid membranes. The third of
the prevalent mutations, H1047R, is located near the
activation loop in the catalytic domain. It is likely that it
affects specificity or affinity of p110 towards the lipid
substrate of PI3K.
Mutated kinases are ideal drug targets especially
when mutated forms of these proteins are only expressed
in tumour cells and result in a gain of function. Studies
of patients with non-small-cell lung cancer have shown
that tumours that express mutant forms of the recep-
tor tyrosine kinase EGFR (epidermal growth factor
receptor) are more responsive to EGFR inhibitors such
as erlotinib and gefitinib, compared with tumours that
express wild-type EGFR148150. Therapy with these inhib-
itors increased survival times of patients with tumours
that express EGFR151.
A rough estimate that is based on the published
frequencies of tumour-specific mutations in PI3K indi-
cates that the number of newly diagnosed patients with
cancer who carry one of the hot spot mutations in
PI3K (described in FIG. 3), is in excess of 10,000 per year
in the United States and there are many more world-
wide. Specific inhibitors of mutant forms of PI3K could
therefore have a significant therapeutic effect. The fea-
sibility of generating mutant-specific PI3K inhibitors
has not been established. However, recent advances in
the design and assembly of bivalent enzyme inhibitors
indicate that compounds with the requisite specificity
and potency could be within reach152154.

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Acknowledgements
The work of the authors is supported by grants from the National
Cancer Institute.
NATURE REVIEWS | C ANCER
Competing interests statement
The authors declare no competing financial interests.
Online links
DATABASES
The following terms in this article are linked online to:
National Cancer Institute: http://www.cancer.gov
breast cancer | colorectal cancer| ovarian cancer | pancreatic
cancer
Swiss-Prot: http://www.expasy.org/sprot
4EBP | AKT | AP1 | BAD | -catenin | caspase 9 | c-MYC |
COT | CRM1 | cyclin D1 | cyclin D2 | DNA-PK | EGFR |
2005 Nature Publishing Group
REVIEWS
ERBB2 | FKBP12 | FOXO1 | FOXO3A | FOXO4 | FOXO6 |
GSK3 | HIF1 | IKK | ILK | MDM2 | MIZ1 | NFB | p21cip1 |
p27kip1 | p53 | PDK1 | PTEN | REL-A | RHEB | S6K | SKP2 |
SMAD | YB-1
FURTHER INFORMATION
Atlas of Genetics and Cytogenetics in Oncology and
Haematology (PIK3CA): http://www.infobiogen.fr/services/
chromcancer/Genes_gc/GC_PIK3CA.html
Peter K. Vogts homepage: http://www.scripps.edu/research/
faculty.php?tsri_id=1847
The PI3KPTENAKT signalling pathway: www.humpath.
com/article.php3?id_article=890
Access to this interactive links box is free online.
VOLUME 5 | DECEMBER 2005 | 929