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Anti-Cancer Drugs: Molecular Mechanisms of Action

Article  in  Mini Reviews in Medicinal Chemistry · August 2005

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 Mini-Reviews in Medicinal Chemistry, 2005, 5, 685-695 685

Anti-Cancer Drugs: Molecular Mechanisms of Action

Mauro Cesar Isoldi*,1 , Maria Aparecida Visconti2 and Ana Maria de Lauro Castrucci1,2

1Department of Biology, Gilmer Hall, University of Virginia, Charlottesville, VA, 22904-4328, USA
2Departamento de Fisiologia, Instituto de Biociências, Universidade de São Paulo, SP, 05508-900, Brazil
Abstract: Genetic alterations are responsible for all cancers. These mutations produce, in turn, alterations in
key proteins of certain signaling pathways. Amongst the best known and studied alterations related to
malignant transformations are those which occur in Ras protein and p53. In most cases mutations in Ras and
p53 lead to the appearance of practically most malignant transformations. Mutated Ras genes exist in
approximately 20 to 30% of all human cancers. Ras proteins are switches that regulate diverse functions such as
cell proliferation, differentiation and apoptosis. Normal p53 expression, also known as the “genome guardian”,
is a key molecule for suppressing cell proliferation.
The great importance of these proteins rests on their intimacy with the events leading to cell proliferation or
death. The comprehension of the extent of transformation on Ras and p53, and of the diverse biochemical
pathways of intracellular signaling, activated by them, is of extreme importance for the understanding of
malignant transformation, as well as its control, through the creation, for example, of new drugs which
contribute to the elimination of these cells.
To clarify the consequences originated by transformed Ras, p53 and their biochemical interlinks in the different
intracellular pathways, besides the possible intervening points and pharmacological controls presently used
in combating cancer, are the aims of this review.
Keywords: Cell signaling, cancer, mechanisms of action, Ras protein, p53.

The advances in our understanding of the basic molecular carcinomas. Thus, aberrant Ras functioning is believed to
mechanisms involved in the control of cell growth, contribute to the development of at least a major subset of
differentiation and survival have provided a framework for the these neoplasms. In contrast mutated ras genes are rarely
pharmacological manipulation of many diseases. associated with the development of breast, ovarian or cervical
carcinomas (< 5%). Therefore, aberrant Ras functioning may
We now know that three types of genetic alterations or
not be important in promoting or maintaining the malignant
mutations underlie the pathogenesis of virtually all cancers.
and invasive properties of these tumors [5].
These mutations arise in oncogenes, tumor suppressor genes
and genes that govern the exact replication of DNA, e.g. The three ras genes encode four closely related 21 kDa
DNA repair enzymes and cellular checkpoint genes [1]. proteins [6] in human cells (Harvey, Kirsten and N-ras), the
Kirsten-ras gene (K-ras) by far the most commonly mutated
Mutations involving the genes of Ras or p53 proteins
form of ras found in human cancers [7]. K-ras encodes two
encompass, in turn, many known malignant transformations.
proteins, K-Ras4A and K-Ras4B, that differ only in their
Our aim in this review is to discuss the main alterations, carboxy-terminal 25 amino acids. K-Ras4B is the
caused by Ras and p53 transformations, and their predominant protein species expressed in mammalian cells
interactions beyond the main intervention points of their [5]. However, all three types of mutated ras genes are capable
intracellular signaling pathways (Fig. 1). of transforming mammalian cells in culture.
Ras proteins function as biological switches that are
Ras PROTEIN AND CANCER regulated by their association with guanine nucleotides
Among the oncogenes associated with human cancers, (GDP and GTP) [8,9] and regulate multiple signal
the ras oncogenes stand out as particularly attractive targets transduction pathways controlling biological processes as
for the creation of cancer therapeutics. Mutated ras genes diverse as cellular proliferation, differentiation and apoptosis
exist in approximately 20-30% of all human cancers but are [10]. Normal Ras proteins typically reside in the inactive
most commonly found in pancreatic cancer, colon cancer and GDP-bound state. Upon initial stimulation at the surface by
adenocarcinoma of the lung [2,3,4]. Frequency is not a variety of extracellular ligands, Ras guanine nucleotide
uniform with respect to tumor type, thus suggesting that ras exchange factors (Sos and RasGRF/CDC25) are activated to
mutation contributes to the development of some, but not all stimulate the exchange of bound GDP for GTP to form the
tumors [5]. For example, ras mutation are highly prevalent active GTP-complexed Ras protein [11]. This active state is
in pancreatic (90%), lung (40%) and colorectal (50%) transient, and Ras GTPase activating proteins (Ras GAPs;
p120 and NFI-GAP) stimulate the intrinsic GTP hydrolysis
of Ras to cycle it back to the inactive GDP-complexed state
*Address correspondence to this author at the Department of Biology, [12].
Gilmer Hall, University of Virginia, Charlottesville, VA, 22904-4328,
USA; Tel: 434-982-5474; Fax: 434-982-5626; E-mail: It is well appreciated that the activation of diverse cell-
[email protected] surface receptors can stimulate convergent signals that lead to

1389-5575/05 $50.00+.00 © 2005 Bentham Science Publishers Ltd.

686 Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 Isoldi et al.

Fig. (1). Ras and p53 pathways. Ras and p53 activate multiple effector pathways. Each of these events triggers a downstream cascade
that leads to the modification of proteins affecting many aspects of cell growth and survival.

activation of Ras [13]. Once activated Ras binds to at least then causes the translocation of Ras to the inner surface of the
three types of effector protein: kinases of the Raf family, plasma membrane [5, 24]. This tetrapeptide sequence is
phosphoinositide (PI) 3-kinase and RalGDS proteins [14]. generally referred to as the CAAX box or CAAX motif, and
consists of cysteine followed by two aliphatic amino acids,
Ras activation of the Raf serine/threonine kinases and the
and any other residue (usually serine or methionine) which is
activation of the ERK mitogen-activated protein kinases
present in all Ras precursor proteins [25-27].
(MAPKs) remain a key signalling important for Ras biology
[13], as it initiates further cytoplasmic and nuclear events to Three enzymatic reactions occur in these residues:
alter cell growth and differentiation [15,16,17]. This pathway prenylation, proteolytic removal and carboxymethylation.
will be mentioned in full detail due to its importance for the Prenylation of the cysteine residue occurs within minutes of
control of malignant transformation. synthesis: the cytoplasmic Ftase catalyzes the covalent
addition of a C15 farnesyl isoprenoid moiety, from a farnesyl
Phosphoinositide (PI) 3-kinases, once activated, promote
pyrophosphate (FPP) donor to the cysteine residue of the
the conversion of phosphatidylinositol (4,5)-bisphosphate
Ras CAAX motif. This modification is rapidly followed by
[PtdIns(4,5)P 2] to phosphatidylinositol (3,4,5)-triphosphate
proteolytic removal of the AAX residues and
[PtdIns(3,4,5)P 3]. One established target of (PI) 3-kinase is
carboxylmethylation of the now farnesylated cysteine residue.
Akt/PKB (protein kinase B) [18] which, among other
H-Ras, N-Ras and K-Ras4A undergo additional modification
functions, promotes cell survival. PI-3kinase may also
of upstream cysteine residue(s) by the fatty acid palmitate,
activate Rac GTPase, and this Rho family protein is an
whereas K-Ras4B contains a lysine-rich sequence directly
important mediator of oncogenic Ras transformation.
upstream of the CAAX motif [25]. Of these three reactions,
Akt/PKB and Rac facilitate activation of the NF-kB
the first and most important for the Ras biological activity is
transcription factor by Ras [19, 20] and exert an anti-
the prenylation of the cysteine residue [28].
apoptotic role in the Ras pathway [21].
Although the CAAX-signaled modifications are essential
The mutant ras genes present in human tumors encode
to promote Ras membrane association, the palmitate
single amino acid mutations (at residues 12, 13 or 61) that
modification or the lysine-rich sequence serves as a second
render these oncogenic Ras mutant proteins insensitive to
membrane-targeting signal to facilitate full plasma membrane
GAP stimulation. Hence, they remain constitutively active
association [25], along with additional sequence information.
in the absence of external stimuli [11, 22].
However, while each carboxy-terminal modification increases
Ras proteins that are encoded by mutated ras genes are protein hydrophobicity and contributes to Ras membrane
unable to cleave GTP, and remain trapped in their GTP association, the initial farnesylation step alone is sufficient to
bound state. The pathophysiological consequence of these promote significant membrane association and transforming
mutations is that Ras driven cell proliferation signal is held potential [29]. Therefore, inhibitors of Ftase may serve as
in the “on” position and inappropriately stimulates potent anti-Ras drugs, as it will be discussed below.
continuous cell proliferation [23]
Ras proteins are initially synthesized as inactive PHARMACOLOGICAL INTERVENTION ON Ras
cytoplasmic proteins. However, a series of rapid post-
translational modifications, signaled by a consensus carboxy- Ras function is regulated by GDP/GTP cycling. This fact
terminal tetrapeptide sequence, present in all Ras proteins, suggests two possible approaches for antagonizing Ras
Anti-Cancer Drugs Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 687

(A) (B)
CH3 CH3 CH3 OH SH
O O
H H
H3C P N N
O H2N OCH3
(α-hidroxyfarnesyl) phosphonic acid O
O
FTI-277

SCH3

SH
(C) O
H H CH3
N N
H2N O O
O CH3

SO2CH3
L-744, 832

Fig. (2). Chemical structures of FTIs. (A) Farnesyl diphosphate analogues; (B) and (C) peptidomimetics (CAAX mimetics).

activity. First, the generation of variant “super” Ras Gaps FTase INHIBITORS (FTIs)
that can stimulate GTPase activity of oncogenic Ras FTIs comprise a novel class of antineoplasic agents
proteins, and thus convert them to inactive proteins, has developed to inhibit FTase [34]. In mammalian, there are
been attempted. Unfortunately, this has been unsuccessful. two prenyltransferases: farnesyltransferase (FTase) and
Second, since the inactive and active forms of Ras differ in geranylgeranyltransferase type I (GGtase). Both enzymes are
protein conformation [11, 22], compounds that preferentially heterodimeric proteins (approximately 94kDa), composed of
recognize the active GTP-bound form may specifically target common alpha sub-units and divergent albeit homologous
mutated Ras proteins. beta sub-units [32,33]. The sequence CAAX box determines
A technically more feasible approach became apparent the process. FTases prefer protein substrates with serine-,
with the finding that Ras requires prenylation to induce methionine- or glutamine-ending CAAX box while GGtases
malignant transformation. Ras must be anchored at the prefer proteins with CAAX boxes that terminate with leucine
plasma membrane to transmit signals through the MAP [35].
kinase pathway. As Ras does not contain a transmembrane The discovery that the tetrapeptide sequence (CAAX
domain, localization is accomplished by the post- sequence) was necessary and sufficient to serve as substrate
translational addition of a lipid (farnesyl) moiety to its for Ftases motivated researchers to design new Ftase
carboxy-terminal. Subsequently, many investigations were inhibitors [36].
initiated exploring farnesyltransferase, the enzyme
responsible for this modification, as an anticancer drug target The FTase inhibitors were of particular interest since the
(Figs. 2 and 3) [30-33]. oncogenic activity of Ras is dependent on its farnesylation

Cl
(A) (B)

N
N
N N H3C
N NH2
O O
H3C N
Cl
N

L-778, 123
R115777
(C)
Cl
Br
N O
Br
NH2
N
N

O SCH66336

Fig. (3). Chemical structures of new generation of FTIs. (A) L-778,123; (B) R115777 and (C) SCH66336.
688 Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 Isoldi et al.

[29]. FTase inhibitors produce reverse oncogenic Raf regulation of cell survival is independent of MEK/ERK
transformation of cell and tumor lines which constitutively pathway, and involves the activation of NF-kB transcription
express active mutant Ras [37]. complex. NF-kB proteins are conserved transcription factors
implicated in the regulation of inflammation,
Initially, farnesyltransferase inhibitors (FTIs) were
morphogenesis, differentiation, proliferation and apoptosis
envisaged as a general mean of impairing the membrane
[52, 53].
localization of oncogenic Ras proteins.
Like acylation with myristate [38] or palmitate [39] and Ras, ARF AND p53
modification with glycosil phosphatidylinositol [40],
prenylation has been viewed as a mechanism for post- Expression of active forms of Ras leads to elevated levels
translational attachment of proteins to membranes [41]. of p53. The mechanism by which Ras induces p53 is not
completely understood. There are three known pathways:
Peptidomimetics of CVIM, the CAAX Box sequence of expression of ARF, of DMP1, a transcription factor, and of
K-Ras4B, were initially used as the first inhibitors of FTases the tumor suppressor protein PML. Ras and Raf promote the
(FTIs) rationally synthesized [36]. At first it was believed expression of ARF [54], which then binds to and promotes
that these peptides could block as much the activity of nucleolar sequestration of Mdm2 [55, 56]. The sequestration
FTase as of the enzyme geranylgeranyltransferase I (GGTase of Mdm2 allows the increase of p53 levels, leading to the
I), which transfers a geranylgeranyl isoprenoid to the protein. induction of target genes which promote cell cycle arrest
Although this is true for H-Ras, the same can not be said in [57].
relation to K-Ras. Subsequent work demonstrated that, in
the presence of FTIs, K-Ras could still be prenylated by the DMP1 is a transcription factor that may induce ARF-
related enzyme GGTase I in vivo [42, 43]. dependent cell cycle arrest. DMP1 is a 120- to 130-kD
nuclear phosphoprotein consisting of 761 amino acids. It is
As K-ras is also a substrate forgeranylgeranyltransferase
composed of a central DNA-binding domain containing three
I, inhibition of farnesyltransferase alone is not sufficient to
Myb-like repeats flanked at the amino- and carboxy-termini
preclude membrane attachment of this oncogenic protein.
by acidic transactivation domains [58, 59]. DMP1 binds to
This finding led investigators to explore the combination of
nonameric consensus DNA sequences [CCCG(G/T)ATGT],
an FTI with a geranylgeranyltransferase inhibitor (GGTI).
and competes with eukaryotic transcriptional regulators (Ets)
This approach was limited by the large number of
(for review, see [60]) for DNA recognition sites that contain a
geranylgeranylated proteins found at the cellular level, some
GGA core. DMP1 is able to physically associate with any of
of which are required for normal cell physiology. Although
the three D-type cyclins, which can inhibit its ability to bind
the development of FTIs was based on the premise that they
DNA with no requirement of CDK4 or CDK6 [59, 61].
would selectively target Ras, there is extensive evidence for
DMP1 can induce the expression of ARF which could
non-Ras proteins being the more relevant target for some
explain its ability to elicit p53-dependent cell cycle arrest
drugs.
and senescence [62].
The inhibition of the Ftase enzyme may rely on
processes other than the known blockade of Ras activity. PML is a RING finger protein localized in large nuclear
Other farnesylated proteins frequently mentioned as target structures called promyelocytic oncogenic domains (PODs),
"X" of FTIs may be more important in this process than Ras ND10, or PML nuclear bodies (for review, see [63]). PML
[44]. Among these novel targets, one has to mention the was initially identified in acute promyelocytic leukemia
members of the Rho family, mainly RhoB [45, 46] and (APL), in which it forms a reciprocal translocation with the
RhoE [47], besides important proteins involved in the cell rar gene [64, 65, 66, 67]. Although the biochemical action of
cycle progression, such as CEMP-AND and CEMP-F PML is not known, it may function in transcription control
(center-mere-binding proteins) [48]. RhoB, an endosomal by recruiting transcription factors to the PODs [68, 69]. Ras
protein related to receptor trafficking, is a protein that, when expression promotes the formation of a ternary complex
non-farnesylated, leads to selective growth inhibition and constituted by p53, PML, and the p300/CBP
apoptosis in tumor cells [49]. acetyltransferase in PODs. Under these circumstances, p53 is
acetylated on lysine residues 320 and 382 what enhances its
We may conclude that the search for farnesylated protein DNA binding activity.
X is far from complete and further efforts must be made to
identify these targets [13].
p53 AND ITS MECHANISM
Ras, Raf AND MEK It has been suggested that the tumour suppressor TP53,
the so called “guardian of the genome” [70], is a direct
Ras interacts with the active form of Raf, a “sensor” of DNA damage. It locates single stranded regions
serine/threonine kinase. The classic pathway regulated by of DNA and termini of non-specific DNA templates [71].
Raf to control apoptosis is directly related to its kinasic
activity. Raf kinases phosphorylate and activate Mek1 and The p53 tumor-suppressor gene encodes a 393-amino
Mek2, both specific kinases. Mek1 and Mek2 in turn acid sequence that resides in the cell nucleus [72], and has
phosphorylate and activate Erk1/p44 and Erk2/p42, the general molecular structure of a transcription factor with a
mitogenic protein kinases which play key roles in the transcriptional activation domain at the amino-terminal (aa:
stimulation of cell division [50, 51]. The molecular steps 1–42), a sequence-specific DNA-binding domain within the
involve transcriptional regulation as well as modifications of central part (aa: 102–292), and an oligomerization domain
pro-apoptotic proteins. Functionally, these pathways can be (aa: 323–356) together with a regulatory domain (aa: 360–
divided in MEK-dependent and MEK-independent cascades. 393) at the carboxy-terminal [73-77].
Anti-Cancer Drugs Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 689

Normal p53 expression suppresses cellular proliferation. auto-regulatory feedback loop between p53 and Mdm2. It is
In response to stressful stimuli (including DNA damage, currently accepted that mutant p53 cannot activate the
hypoxia, radiation, oxidative stress, heat shock, metabolic transcription of Mdm2, and for this reason p53 accumulates
changes, nucleotide depletion, or exposure to certain within the nucleus. Moreover, in some tumors, p53 is
cytokines), the p53 gene is activated, binds in tetrameric inactivated by over-expression of Mdm2 rather than by
form [78, 79] to four palindromic copies of its consensus mutation. In addition, the INK4a-ARF locus encodes two
sequence [73], and its protein accumulates in the nucleus, proteins—p16 (INK4A) and p14, which affect the function of
triggering downstream cascades that terminate in cell cycle the retinoblastoma susceptibility gene (Rb) protein and p53,
arrest or apoptosis. respectively. p14 inhibits cell cycle progression by
promoting Mdm2 degradation and stabilizing wild-type p53.
Therefore, the activation of p53 is central to the fate of a
p16 (INK4A) is a member of a family of specific inhibitors
cell that encounters a hostile environment [80]. p53 also
for CDK4 and CDK6 [89].
functions as a transcription factor that can bind to specific
DNA sequences and activate the transcription of genes Two independent events describe the instability of the
containing binding sites in their promoter regulatory regions complex constituted by p53 and Mdm2. The first is the
[80]. phosphorylation of serine residues in the amino- and
carboxy-termini of p53 [90]. The second and main
Although under certain circumstances trans-regulatory-
mechanism involves the reduction of MDM2 expression in
independent mechanisms may be of significance for p53
response to genomic stress. Mutations involving
function, there is compelling evidence that trans-regulation
phosphorylation sites of p53 do not alter the expected
of downstream genes is essential for this action, especially
activation of this molecule after genotoxic stress [74].
for tumor suppression [81].
Subsequent work demonstrated however that, in spite of the
p53 transcriptionally activates a number of key enzymes relevance of p53 phosphorylation for a fast activation of the
mediating the pathway leading to cell cycle arrest or molecule, the maintenance of high levels of p53 expression
apoptosis, including p21/WAF1/Cip1 (which arrests the cell by ARF, whose expression is controlled by E2F, is also
cycle by inhibiting cyclin-dependent kinase complexes and very important [91].
binding to proliferating cell nuclear antigen [PCNA]), Bax
The DNA binding sites of p53 protein are located inside
(which promotes apoptosis), GADD45 (which arrests the cell
three loop-helix structures [92, 93]. Although different from
cycle by binding to PCNA), and insulin-like growth factor-
the classic "zinc fingers", these loops are connected and
binding protein 3 (which enhances apoptosis by blocking the
stabilized by a divalent atom of zinc [94]. The p53
mitotic activity of insulin-like growth factors) [82].
homologue proteins, p73 and p63, also present this structure
In addition, p53 helps to regulate angiogenesis, since it and mutations at this region account for 80% to 90% of the
functions as a transcription factor for the vascular endothelial malignant transformations [94].
growth factor (VEGF) [83], the basic fibroblast growth factor
The actual site of the mutation is important, since
(bFGF) [84], thrombo-spondin [85], and a thrombospondin-
mutations in the DNA-binding domain have the greatest
like factor, brain-specific angiogenesis inhibitor [86]. Wild-
effect on function. Most missense mutations in cancers are
type p53 downregulates endogenous VEGF and bFGF
located in this domain, and mutations here lead to the
production, thereby limiting tumor growth by limiting the
production of a p53 protein that fails to bind to DNA in the
induction of neovascularization caused by the overproduction
normal sequence-specific fashion.
of these angiogenic factors [85, 86]. Mutant p53 loses this
important regulatory function, allowing neovascularization to The carboxy-terminus of p53 regulates its binding to
proceed uninhibited. DNA in a negative way [77]. The amino-terminal seems to
participate in a cooperative manner: free peptides
The number of downstream genes trans-repressed and
corresponding to p53 carboxy-terminus (aa 361-382) bind to
trans-activated by p53 is presently estimated at 70 and 80,
proline-rich regions (aa 80-93) of the amino-terminal [95].
respectively [74].
This binding promotes conformational changes in the p53
In addition, p53 is regulated by another nuclear protein, molecule, leading the cell to apoptosis [95]; via Fas
MDM2, which modulates the observed accumulation of p53 (APO1)/Fas ligand pathway [95].
in response to stress. MDM2 is a 491-amino acid
Following genotoxic stress, p53 protein rapidly
phosphoprotein that binds to p53, blocking its biological
accumulates and becomes activated. The kinetics and the
activity. MDM2 also targets p53 for destruction by way of
duration of p53 accumulation can vary considerably in
the ubiquitin proteosome pathway [80].
different cell types in response to various damaging agents
The Mdm2 gene itself, on the other hand, is trans- [89]. p53 is widely recognized as a protein functional during
activated by p53, resulting in a negative feedback control of the cell cycle. Cell cycle arrest at G1 phase by p53 is
p53 activity [87]. dependent upon the transcriptional activation of
It has been hypothesized that phosphorylation of Mdm2 p21/WAF1/CIP1 [96, 97]. p21 is a cyclin-dependent kinase
or p53 itself by stress-activated protein kinases prevents the (CDK) inhibitor. Rb protein phosphorylation is mediated by
interaction between Mdm2 and p53, and hence allows the CDKs. The hypophosphorylated Rb sequesters the
accumulation of p53. transcription factors of the E2F family known to promote the
S phase [98].
However, high p53 levels were shown to trans-repress
Mdm2 gene activity through promoter 3 located in intron 3 The p53-promoted blockade in G2 involves inhibition of
of this gene [88]. This finding adds a further element to the Cdc2, a cyclin-dependent kinase required for the entrance in
690 Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 Isoldi et al.

mitosis. Cdc2 is inhibited by three transcriptional targets of MEK-ERK pathway can elicit strikingly different effects on
p53: Gadd45, p21 and the 14-3-3 sigma [99], although some the cell cycle depending on the level of activation [110,
data also indicate that 14-3-3 sigma is mainly under the 111].
transcriptional control of the p53 homologue p73 [100].
p53-dependent apoptosis is not so well understood. Bax, p53, Ras AND CELL CYCLE
which is known to antagonize the anti-apoptotic activity of
Bcl-2, seems to be the mediator used by p53 in its apoptotic The cell cycle inhibitory effects of Ras, or its effector
activity [101, 102]. The activation of BAX gene by JMY protein kinase, Raf, have been described in a variety of cells
(p300 cofactor) in the p300/CBP (CREB-binding protein) [112, 113]. This is likely to be achieved by the ability of
transcriptional co-activator multi-protein complexes, which Ras to infuence the activity of cyclin-dependent kinases
present variations in variant splices of JMY, determines, at (CDKs) that phosphorylate pRb and its family members
least in part, whether p53 is going to mediate the cell cycle [114, 115]. The phosphorylation of pRb promotes the
arrest or apoptosis [103].The activation of caspase cascade activity of those E2F transcription factors required for the
seems to be another way for p53-induced apoptosis. expression of genes that promote S-phase progression [116,
Members of the TNF receptor superfamily, especially 117]. In contrast to promote cell cycle progression, Ras and
Faz/APO1 and TRAIL/DR5, were also discovered to be its various effectors also have the ability to elicit cell cycle
involved in p53-dependent apoptosis. The activation of these arrest and, in the case of primary cells, to induce a form of
receptors rapidly leads the cells to apoptosis mediated by irreversible arrest that has features of replicative senescence
caspase cascade [95, 104, 105]. This pathway is also [118, 119].
triggered when p53 regulates enzyme homologues of quinone Ras activation, working through Raf, can induce the
oxidoreductase, proline oxidase and glutathione transferase. expression of CDK inhibitors (CKIs) of the INK4 or Cip/Kip
These enzymes induce apoptosis by producing reactive family [120, 121]. CKI induction inhibits the activity of
oxygen intermediates (ROI) with subsequent mitochondrial CDKs leading to arrest in the G1 and/or G2 phases of the
damage and caspase activation [106]. cell cycle.

Mdm2 EXPRESSION BY Ras AND Raf NEW BIOCHEMICAL TARGETS

The induction of p53 by a variety of agents leads to the Other biochemical pathways, besides those activated by
transcriptional induction of Mdm2 which, in turn, promotes p53 and Ras, have been explored to design anticancer drugs.
p53 degradation [107, 108]. However, Mdm2 is also Among them, the serine/threonine kinases such as protein
induced by mitogenic stimulation or oncogenic kinases C (PKCs), cAMP-dependent protein kinases (PKAs)
transformation of cells [109, 110]. Mdm2 gene is regulated and Auroras, besides proteases, as the caspases, have been
by Ras-induced Raf/MEK/MAP kinase pathway, in a p53- mostly investigated.
independent manner. Raf-induced Mdm2 degrades p53 in the
absence of the Mdm2 inhibitor p19ARF. This regulatory PKC exists as a family of at least 12 closely related
pathway accounts for the observation that cells transformed isozymes [122]. Each isoform plays different roles in cell
by oncogenic Ras are more resistant to p53-dependent growth, proliferation, differentiation or apoptosis [123]. Due
apoptosis originated from DNA damage [110]. to their important roles in many different cancers, PKCs
could be potential targets for developing novel cancer
Activation of Ras-induced Raf/MEK/MAP kinase may therapies. Treatments utilizing antisense oligonucleotides
therefore play a key role in suppressing p53 during tumor directed against PKC-alpha are in clinical development for
development and treatment. Indeed it is clear that the Raf- several cancers [124, 125]. Bryostatin, a compound that

(A) OH
CH3 (B)
O
Cl
O CH3 Cl
Br N
O
S
S NH S
H3C
O
BH3 l-1 OH O
Cl
BH3 l-2
(C)
CH3 CH3 O
+
O N
H3C O
O

Chelerythrine

Fig. (4). Chemical structures of Bcl2 inhibitors. (A) BH3 I-1; (B) BH3 I-2 and (C) chelerythrine.
Anti-Cancer Drugs Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 691

inhibits PKC-alpha is a good example [126-128]. Utilization members of the human Aurora kinases are over-expressed in
of inhibitors of other PKC isoforms has not yet been tested a variety of human cancers (for a full review see [138]). This
[129]. fact, added to the involvement of Aurora kinases in mitosis,
opens a great area of studies for the design of new drugs to
PKA is involved in controlling cell growth and
control their function.
differentiation [130], and is present in mammalian cells in
two distinct isoforms: PKAI and PKAII [131]. The PKAI Apoptosis is mediated by proteases called caspases.
isoform is over-expressed in human cancer and is directly Perturbed regulation of apoptosis underlies many diseases
involved in EGFR mitogenic signaling [132]. PKAI has including cancer. The link between selective cell suicide and
been proposed as a relevant target for cancer therapy. Down- cancer emerged when Bcl-2 was found to inhibit cell death
regulation of PKAI by pharmacological tools, including [139]. Bcl2 and its homologues, Bcl-x1 and Bcl-w, potently
antisense oligodeoxynucleotides targeting its RI alpha inhibit apoptosis in response to many cytotoxic insults
subunit (AS-PKAI) causes growth arrest and differentiation in [140]. The Bcl-2 family members regulate apoptosis and
a variety of cancer cell lines in vitro and in nude mice [131, mitochondrial integrity [141], and represent key targets for
133]. therapeutic intervention [142]. Small molecules have been
used as inhibitors of the Bcl2 family, for example BH3 I-1
The Aurora kinases have essential functions in cell
and BH3 I-2 [143], and chelerythrine [144] (Fig. 4).
division [134, 135]. In mitosis, Aurora kinases are required
for chromosome segregation, condensation and orientation in
the metaphase plate, spindle assembly, and the completion PHARMACOLOGICAL INTERVENTION ON
of cytokinesis. Three mammalian Aurora kinases appear at TOPOISOMERASE AND p53
specific locations during mitosis. Aurora-A, the "polar
kinase", primarily associates with the separating centrosomes DNA topoisomerases are critical enzymes involved in
while Aurora-B, the "equatorial kinase", is a chromosomal replication, transcription, chromatin assembly and other
passenger protein [134], and Aurora-C is localized to the aspects of DNA metabolism. All cells have two major forms
centrosome from anaphase to telophase [136, 137]. All three of topoisomerases: the type I enzymes, that make single-

(A) (C)
O OH O R1
O
O O
R
OH
OH O
OH
OCH3 O OH O O
O
O O
Me
O
NH2
OH
Doxorubicin OH
H3CO OCH3
Daunorubicin H
R2
(B)

R2 R1 O R1 R2
Ctoposide CH3 OH
R3
N O
Etoposide
Phosphate CH3 OPO3H2
R4 N O
OH Teniposide S OH
H3C

Topotecan R2 CH2-N(CH3 )2 .HCl (D)

R3 OH
R
SN-38 R1 C 2H5 H
N
R3 OH

R 2-methoxy- 4-methanesulfonamido

'amsacrine'

Fig. (5). Chemical structures of (A) anthracyclins; (B) camptothecins; (C) epipodophyllotoxins; (D) amsacrine.
692 Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 Isoldi et al.

stranded cuts in DNA, and the type II enzymes, that cut and thereby creating a single stranded DNA break. Cleavable
pass double-stranded DNA. The type II topoisomerases are complexes are also formed in the vicinity of DNA lesions
specific targets of classes of drugs that comprise complex- and in the presence of the anti-tumor agent, camptothecin
stabilizing (epipodophyllotoxins, anthracyclines) (Fig. 5) [156].
and catalytic (merbarone, bisdioxopiperazines) (Fig. 6) While many agents appear to inhibit the catalytic activity
inhibitors [145]. of topoisomerase I, most bind to DNA very tightly, and
O have little specificity for topoisomerase I (berenyl and
O O
H3C H O ethidium bromide) (Fig. 7) [157]. However, drugs that bind
N HN N the minor groove of DNA may be more specific inhibitors of
N
N NH topoisomerase I, although many minor groove binding drugs
S N O O H CH3 also stabilize a covalent complex between DNA and
O topoisomerase I.
ICRF-193 The catalytic cycle of topoisomerase II has been shown to
Merbarone
involve several discrete steps, including recognition/binding,
Fig. (6). Chemical structures of merbarone and ICRF-193 (an cleavage, strand passage, religation, and enzyme turnover
example of bisdioxopiperazine). [158].
Topoisomerase II has also been shown to bind the The anti-tumor importance of the catalytic cycle of
regulatory sequences of the carboxy-terminal region. A topoisomerase relates to the fact that the clinically useful
substantial role in the induction of apoptosis by drugs (etoposide, doxorubisin, amsacrine, toporecan, etc)
topoisomerase II inhibitors such as etoposide and (Fig. 5) appear to be able to increase the amount of species
doxorubicin, which cause accumulation of p53-interacting that include the enzyme covalently attached to DNA, thereby
enzyme–DNA adducts has been suggested [146]. In turning the enzyme into a cell poison, leaving the DNA
mammalian cells, there are two isozymes of topoisomerase strands broken [153, 154, 158]. Drugs accomplish this by
II, a 170 kDa form termed p170, or alfa, and a 180 kda form blocking the religation of the cleaved DNA, or by increasing
termed p180, or beta [147, 148]. These two proteins are the rate of DNA cleavage without inhibiting religation [154].
products of different genes, located in human chromosomes For both biochemical mechanisms, drugs stabilize a
17q21-23 [149] and 3q [150], respectively. These isoforms “cleaved complex” of DNA and topoisomerase.
are differentially expressed through the cell cycle: Consequences of the formation of a covalent complex with
topoisomerase II alpha is preferentially expressed in DNA include interference with nucleic acid metabolism, the
proliferating cells during S phase [151], whereas induction of genetic changes due to the presence of a DNA
topoisomerase II beta appears to be expressed at all points in strand break, and the initiation of an apoptotic pathway.
the cell cycle, with no appreciable differences between
proliferating and non-proliferating cells [152].
DRUG RESISTANCE
Topoisomerases have been shown to be targets of
clinically important anti-tumor agents [153, 154]. For The majority of anti-tumor drugs damage DNA, either
example, topoisomerase I is a very specific target of directly or indirectly. The idea that this damage per se is not
camptothecin and its analogues such as topotecan, 9-amino- lethal but has to be “sensed” by the cell and coupled to the
camptothecin, irinotecan (CPT-11) [155] and execution of apoptosis suggests that failure of the “sensors”
indolocarbazoles (rebeccamycin and staurosporine) which could lead to drug resistance (as well as promotion of
also inhibit PKC [95]. carcinogenesis). As mentioned above, p53 has been
described as a “sensor” of DNA damages.
Mammalian DNA topoisomerase I is a multifunctional
enzyme which is essential for embryonic development. Mutations in p53 have been found to be associated with
Topoisomerase I is a recombinase, which can mediate drug resistance in vivo as well as in animal models [159,
illegitimate recombination. A crucial intermediate reaction 160, 161]. Epidemiological studies of p53 mutations and
during relaxation of DNA is the formation of a DNA- drug responsiveness yielded controversial results in a large
topoisomerase I complex (the cleavable complex) wherein variety of tumors treated with different regimens of cytostatic
topoisomerase I is covalently linked to the 3’-end of DNA, drugs [162-165]. Most trials show an association between

NH2

H
N N
N NH2 -
H2N N+ Br
H2N
NH
NH

Berenil Ethidium Bromide

Fig. (7). Chemical structures of berenil and ethidium bromide.

Anti-Cancer Drugs Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 693

p53 mutation and chemotherapy resistance when dealing ACKNOWLEDGEMENTS

with hematological malignancies. The influence of p53 on This work has been partially supported by FAPESP and
sensitivity to anticancer agents is, however, by no means CNPq.
clear in malignant tissues of non-hematological origin. The
significance of the mutation of p53 and of allelic loss to the
progress of human neoplasia as an indicator of poor REFERENCES
prognosis, is unquestionable. Whether the loss of p53 [1] Oliff, A. Biochim. Biophys. Acta , 1999, 1423, C19-C30.
function alone is responsible for pleiotropic drug resistance [2] Bollag, G.; McCormick, F. Annu. Rev. Cell Biol., 1991, 7, 601-632.
to DNA damaging drugs observed in many advanced [3] Lowy, D.R.; Willumsen, B.M. Ann. Rev. Biochem., 1993, 62, 851 -
cancers, is doubtful [166]. 891.
[4] Clark, G.J.; Der, C.J. Breast Cancer Res. Treat., 1995, 1,133-44.
Resistance to topoisomerase II inhibitors can manifest as [5] Cox, A.D.; Der, C.J. Biochim. Biophys. Acta , 1997, 1333, F51-71.
the decreased or increased expression or mutation of the [6] Barbacid, M. Annu. Rev. Biochem., 1987, 56, 779-827.
topoisomerase II genes. However, response of the tumor cell [7] Sinn, E.; Muller, W.; Pattengale, P.; Tepler, I.; Wallace, R.; Leder,
P. Cell, 1987, 49, 465-475.
to these inhibitors involves more than the target enzyme. [8] Bourne, H.R.; Sanders, D.A.; McCormick, F. Nature, 1990, 349,
Such cell changes are associated with, and may contribute 117-126.
to, the drug resistance phenotype. They involve decreased [9] Quilliam, L.A.; Khosravi-Far, R.; Huff, S.Y.; Der, C.J. BioEssays,
drug accumulation due to expression of membrane “pump” 1995, 17, 395-404.
[10] Reuther, G.W.; Der, C.J. Curr. Opin. Cell Biol., 2000, 12, 157-165.
proteins, altered cytotoxic signaling through stress-activated [11] Tong, L.; de Vos, A.M.; Milburn. M.V.; Jancarik, J.; Noguchi, S.;
protein kinases, and alterations in apoptosis and cell cycle Nishimura, S.; Miura, K.; Ohtsuka, E.; Kim, S.H. Nature, 1989,
proteins (e.g. Bcl-2, Bax, p53, Rb). While it is evident that 337, 90-93.
mutation or altered expression of the topoisomerase II genes [12] Boguski, M.S.; McCormick, F. Nature, 1993, 366, 643-654.
are sufficient to confer resistance to topoisomerase inhibitors, [13] Shields, J.M.; Pruitt, K.; McFall, A.; Shaub, A.; Der, C.J. Trends
Cell Biol., 2000, 10, 147-154.
it is not clear whether the other changes are a consequence of [14] McFall, A.; Ulku, A.; Lambert, Q.T.; Kusa, A. Rogers-Graham,
the selection or a response to the cytotoxic insult, nor is it K.; Der, C.J. Mol. Cell Biol., 2001, 16, 5488-5499.
clear how these other cellular changes contribute to the drug [15] Davis, R.J. J. Biol. Chem. 1993, 268, 14553-14556.
resistance phenotype [145]. [16] Egan, S.E.; Weinberg, R.A. Nature, 1993, 365, 781-783.
[17] Marshall, C.J. Cell, 1995, 80, 179-185.
Because some topoisomerase inhibitors (etoposide, [18] Khwaja, A. Nature, 1999, 401, 33-34.
doxorubicin) are substrates for P-glycoprotein (Pgp), cells [19] Romashkova, J.A.; Makarov, S.S. Nature, 1999, 401, 86-90.
[20] Ozes, O.N.; Mayo, L.D.; Gustin, J.A.; Pfeffer, S.R.; Pfeffer, L.M.;
that express this efflux pump protein will display resistance Donner, D.B. Nature, 1999, 401, 82-85.
to these agents due to decreased drug accumulation [167, [21] Mayo, M.W.; Wang, C.Y.; Cogswell, P.C.; Rogers-Graham, K.S.;
168]. Cells expressing Pgp have been shown to be modestly Lowe, S.W.; Der, C.J.; Baldwin, A.S. Jr. Science, 1997, 278,
resistant to the camptothecin analogue, topotecan, a likely 1812-1815.
consequence of this drug being a substrate for Pgp, other [22] Krengel, U.; Schlichting, L.; Scherer, A.; Schumann, R.; Frech,
M.; John, J.; Kabsch, W.; Pai, E.F.; Wittinghofer, A. Cell, 1990,
camptothecin analogues being poor substrates for Pgp [169]. 62, 539-548.
Renal cell carcinomas exhibit strong resistance to most [23] Bollag, G.; McCormick, F. Cancer Biol., 1992, 3, 199-208.
chemotherapeutic treatments probably due to the expression [24] Gibbs, J.B.; Oliff, A. Annu. Rev. Pharmacol. Toxicol., 1997, 37,
of various multidrug resistance genes. Over-expression of 143-166.
[25] Hancock, J.F.; Magee, A.I., Childs, J.E.; Marshall, C.J. Cell, 1989,
Pgp is established as one such factor [170]. 57, 1167-1177.
Similarly, cells that can also express MRP, the [26] Hancock, J.F.; Paterson, H.; Marshall, C.J. Cell, 1990, 63, 133-
multidrug resistance associated protein, are resistant to 139.
[27] Hancock, J.F.; Hall, A. EMBO J., 1993, 5, 1915-21.
etoposide and doxorubicin [171]. While both Pgp (MDR1) [28] Willingham, M.C.; Pastan, I.; Shih, T.Y.; Scolnick, E.M. Cell,
and MRP have been shown to be expressed in tumor therapy 1980, 19, 1005-1014.
resistant patients [172, 173], the importance of these [29] Kato, K.; Cox, A.D.; Hisaka, M.M.; Graham, S.M.; Bus, J.E., Der,
resistance associated proteins has yet to be fully explored in C.J. Proc. Natl. Acad. Sci. U.S.A., 1992, 89, 6403-6407.
terms of clinical resistance to inhibitors of topoisomerase. [30] Reiss, Y.; Goldstein, J.L.; Seabra, M.C.; Casey, P.J.; Brown, M.S.
Cell, 1990, 62, 81-88.
[31] Gibbs, J.B.; Pompliano, D.L.; Mosser, S.D.; Rands, E.; Lingham,
CONCLUSIONS R.B.; Singh, S.B.; Scolnick, E.M.; Kohl, N.E.; Oliff, A. J. Biol.
Chem., 1993, 268, 7617-7620.
The advance in knowledge of intracellular biochemical [32] Chen, W.J.; Andres, D.A.; Goldstein, J.L.; Russell, D.W.; Brown,
alterations produced by malignant transformation has led M.S. Cell, 1991, 66, 327-341.
man closer to the definitive control of cancer, by developing [33] Omer, C.A.; Kral, A.M.; Diehl, R.E.; Prendergast, G.C.; Powers,
S.; Allen, C.M.; Gibbs, J.B.; Kohl, N.E. Biochemistry, 1993, 19,
more efficient, less toxic and more specific drugs. 5167-5176.
Domination of these mechanisms has led us to dream of a [34] Kohl, N.E. Ann. N.Y. Acad. Sci., 1999, 886, 91-102.
future, where we can treat cancer bearers according to the [35] Moores, S.L.; Schaber, M.D.; Mosser, S.D.; Rands, E.; O'Hara,
neoplasic mutations encountered. The manipulation of gene M.B.; Garsky, V.M.; Marshall, M.S.; Pompliano, D.L.; Gibbs, J.B.
J. Biol. Chem., 1991, 266, 14603-14610.
alteration has become reality with the use of viral vectors and [36] Cox, A.D. Drugs, 2001, 61, 723-732.
other agents capable of introducing “new commands” in [37] Lobell, R.B.; Kohl, N.E. Cancer Metastas. Rev., 1998, 17, 203-
altered cells, re-doing alterations or modulating them, thus 210.
drawing the respective biological system closer to its normal [38] Boutin, J.A. Cell Signal., 1997, 9, 15-35.
standards. Following this line of thought, the combination [39] Dumphy, J.T.; Linder, M.E. Biochim. Biophys. Acta, 1998, 1436,
245-261.
of gene therapy with other forms of treatment represents one [40] Takeda, J.; Kinoshita, T. Trends Biochem. Sci., 1995, 20, 367-371.
of the most promising options for the future of cancer cure.
694 Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 Isoldi et al.

[41] Glomset, J.A.; Gelb, M.H.; Farnsworth, C.C. Trends Biochem. Sci, [81] Zeimet, A.G.; Riha, K.; Berger, J.; Widschwendter, M.; Hermann,
1990, 15, 139-142. M.; Daxenbichler, G.; Marth, C. Biochem. Pharmacol., 2000, 60,
[42] Rowell, C.A.; Kowalczyk, J.J.; Lewis, M.D. J. Biol. Chem., 1997, 1153-1163.
272, 14093-14097. [82] Steele, V.E.; Wyatt, G.P.; Kellof, G.J.; Elmore, E. Anticancer
[43] Whyte, D.B.; Kirchmeier, P.; Hockenberry, T.N.; Nunez-Oliva, Res., 1998, 6A, 4067-4070.
I.; James, L.; Catino, J.J.; Bishop, W.R.; Pai, J.K. J. Biol. Chem. [83] Kieser, A.; Weich, H.A.; Bradner, G.; Marme, D.; Kolch, W.
1997, 272, 14459-14464. Oncogene, 1994, 9, 963-969.
[44] Kloog, Y.; Cox, A.D. Mol. Med. Today , 2000, 10, 398-402. [84] Ueba, T.; Nosaka, T.; Takahashi, J.A.; Shibata, F.; Florkiewicz,
[45] Sebti, S.M.; Hamilton, A.D. Oncogene, 2000, 56, 6584-93. R.Z.; Vogelstein, B. Oda, Y.; Kikuchi, H. Hatanaka, M. Proc.
[46] Prendergast, G.C. Nat. Rev. Cancer , 2001, 1162-1168. Natl. Acad. Sci. U.S.A., 1994, 91, 9009-9013.
[47] Haluska, P.; Dy, G.K.; Adjei, A.A. Eur. J. Cancer, 2002, 38, [85] Dameron, K.M.; Volpert, O.V.; Tainsky, M.A.; Bouck, N. Cold.
1685-1700. Spring Harbor. Symp. Quant. Biol., 1994, 59, 483-489.
[48] Ashar, H.R.; James, L.; Gray, K.; Carr, D.; Black, S.; Armstrong, [86] Nishizaki, M.; Fujiwara, T.; Tanida, T.; Hizuta, A.; Nishimori, H.;
L.; Bishop, W.R.; Kirschmeier, P. J. Biol. Chem., 2000, 275, Tokino, T.; Nakamura, Y.; Bouvet, M.; Roth, J.A.; Tanaka, N.
30451-30457. Clin. Cancer. Res., 1999, 5, 1015-1023.
[49] Herrera, R.; Sebolt-Leopold, J.S., Trends Mol. Med., 2002, 8, S27- [87] Wu, G.S.; Burns, T.F.; McDonald, E.R.; Jiang, W.; Meng, R,
S31. Krantz, I.D.; Kao, G.; Gan, D.D.; Zhou, J.Y.; Muschel, R.;
[50] Kyriakis, J.M.; App, H.; Zhang, X.F.; Banerjee, P.; Brautigan, Hamilton, S.R.; Spinner, N.B.; Markowitz, S.; Wu, G.; el-Deiry,
D.L.; Rapp, U.R.; Avruch, J. Nature, 1992, 358, 417-421. W.S. Nat. Genet., 1997, 17, 141-143.
[51] Chong, H.; Vikis, H.G.; Guan, K.L. Cell Signal., 2003, 15, 463- [88] Liang, H.; Lunec, J. A.A.C.R. J. Serv.,1999, 40, 701.
469. [89] Shapiro, G.I.; Edwards, C.D.; Rollins, B.J. Cell. Biochem.
[52] Jia, S.; Flores-Saaib, R.D.; Courei, A.J. Mol. Cell Biol., 2002, 22, Biophys., 2000, 2, 189-97.
5089-5099. [90] Shieh, S.Y.; Ikeda, M.; Taya, Y. Cell, 1997, 91, 325-334.
[53] Lin, L.; DeMartino, G.N.; Greene, W.C. EMBO J., 2000, 19, [91] Ashcroft, M.; Vousden, K.H. Onconege, 1999, 53, 7637-7643
4712-4722. [92] Hainaut, P.; Milner, J. A. Cancer Res., 1993, 53, 1739-1742.
[54] Quelle, D.E.; Zindy, F.; Ashmun, R.A.; Sherr, C.J. Cell, 1995, 83, [93] Hainaut, P.; Milner, J. Cancer Res, 1993, 53, 4469-4473.
993-1000. [94] Kaelin, W.G. Jr. Science, 1998, 281, 57-58.
[55] Sherr, C.J.; Weber, J.D. Curr. Opin. Genet. Dev., 2000, 10, 94-99. [95] Kim, A.L.; Raffo, A.J.; Brandt-Rauf, P.W.; Pincus, M.R.; Monaco,
[56] Weber, J.D.; Taylor, L.J.; Roussel, M.F.; Sherr, C.J.; Bar-Sagi, D. R.; Abarzua, P.; Fine, R.L. J. Biol. Chem., 1999, 274, 34924-
Nat. Cell Biol., 1999, 1, 20-26. 34931.
[57] Chan, T.A.; Hwang, P.M.; Hermeking, H.; Kinzler, K.W.; [96] el-Deiry, W.S.; Tokino, T..; Velculescu, V.E.; Levy, D.B.;
Vogelstein, B. Genes Dev., 2000, 14, 1584-1588. Parsons, R.; Trent, J.M.; Lin, D.; Mercer, W.E.; Kinzer, K.W.;
[58] Hirai, H.; Sherr, C.J. Mol. Cell. Biol., 1996, 16, 6457-6467 Vogelstein, B. Cell, 1993, 75, 817-825.
[59] Inoue, K.; Sherr, C.J. Mol. Cell Biol., 1998, 18, 1590-1600. [97] Agarwal, S.; Mathur, M.; Shukla, N.K.; Ralhan, R. Oral Oncol.,
[60] Oikawa, T. Cancer Sci., 2004, 95, 626-633. 1998, 34, 353-360.
[61] Inoue, K.; Sherr, C.J.; Shapiro, L.H. J. Biol. Chem., 1998, 273, [98] Sherr, C.J. Genes Dev., 1998, 12, 2984-2991.
29188-29194. [99] Taylor WR, Stark GR. Oncogene, 2001, 15, 1803-1815.
[62] Inoue, K.; Roussel, M.F.; Sherr, C.J. Proc. Natl. Acad. Sci. U.S.A., [100] Yu, J.; Zhang, L.; Hwang, P.M.; Rago, C.; Kinzler, K.W.;
1999, 96, 3993-3998. Vogelstein, B. Proc. Natl. Acad. Sci. U.S.A., 1999, 96, 14517-
[63] Zhong, S.; Salomoni, P.; Pandolfi, P.P. Nat. Cell Biol., 2000, 2, E85- 14522.
E90. [101] Rao, L.; White, E. Curr. Opin. Genet. Dev., 1997, 7, 52-58.
[64] de The, H.; Lavau, C.; Marchio, A.; Chomienne, C.; Degos, L.; [102] McCurrach, M.E.; Connor, T.M.; Knudson, C.M.; Korsmeyer, S.J.;
Dejean, A. Cell, 1991, 66, 675-684. Lowe, S.W. Proc. Natl. Acad. Sci. U.S.A., 1997, 94, 2345-2349.
[65] Goddard, A.D.; Borrow, J.; Freemont, P.S.; Solomon, E. Science, [103] Shikama, N.; Lee, C.W.; France, S; Delavaine, L.; Lyon, J.;
1991, 254, 1371-1374. Krstic-Demonacos, M.; La Thangue, N.B. Mol. Cell, 1999, 4, 365-
[66] Kakizuka, A.; Miller, W.H.J.; Umesono, K.; Warrell, R.P.J.; 376.
Frankel, S.R.; Murty, V.V.; Dmitrovsky, E.; Evans, R.M. Cell, [104] Eischen, C.M.; Leibson, P.J. Adv. Pharmacol., 1997, 41, 107-132.
1991, 66, 663-674. [105] Wu, G.S.; Burns, T.F.; McDonald, E.R.; Meng, R.D.; Kao, G.;
[67] Kastner, P.; Perez, A.; Lutz, Y.; Rochette-Egly, C.; Gaub, M.P.; Muschel, R.; Yen, T.; el-Deiry, W.S. Oncogene, 1999, 18 , 6411-
Durand, B.; Lanotte, M.; Berger, R.; Chambon, P. EMBO J., 1992, 6418.
11, 629-642. [106] Polyak, K.; Xia, Y.; Zweier, J.L.; Kinzler, K.W.; Vogelstein, B.;
[68] LaMorte, V.J.; Dyck, J.A.; Ochs, R.L.; Evans, R.M. Proc. Natl. A. Nature, 1997, 389, 300-305.
Acad. Sci., 1998, 95, 4991-4996. [107] Fuchs, S.Y.; Adler, V.; Buschmann, T.; Wu, X.; Ronai, Z.
[69] Doucas, V.; Tini, M.; Egan, D.A.; Evans, R.M. Proc. Natl. Acad. Oncogene, 1998, 17, 2543-2547.
Sci. USA, 1999, 96, 2627-2632. [108] Honda, R.; Yasuda, H. EMBO J., 1999, 18, 22-27.
[70] Lane, D.P. Nature, 1992, 358, 15-16. [109] Shaulian, E.; Resnitzky, D.; Shifman, O.; Blandino, G.;
[71] Bavle, I.H.; Elenbass, B.; Levine, A.J. Proc. Natl. Acad. Sci. Amsterdam, A.; Yayon, A.; Oren, M. Oncogene, 1997, 15, 2717 -
U.S.A., 1995, 92, 5729-5733. 2725.
[72] Fujimoto, K.; Yamada, Y.; Okajima, E.; Kakizoe, T.; Sasaki, H.; [110] Ries, S.; Biederer, C.; Woods, D.; Shifman, O.; Shirasawa, S.;
Sugimura, T.; Terada, M. Cancer Res., 1992, 52, 1393-1398. Sasazuki, T.; McMahon, M.; Oren, M.; McCormick, F. Cell, 2000,
[73] el-Deiry, W.S.; Kern, S.E.; Pietenpol, J.A.; Kinzler, K.W.; 103, 321-330.
Vogelstein, B. Nat. Genet., 1992, 1, 45-49. [111] Cook, S.J.; Aziz, N.; McMahon, M. Mol. Cell Biol., 1999, 19, 330 -
[74] Matlashewski, G. Oncogene, 1999, 18, 7618-7620. 341.
[75] Fields, S.; Jang, S.K. Science, 1990, 249, 1046-1049. [112] Bailleul, B.; Surani, M.A.; White, S.; Barton, S.C.; Brown, K.;
[76] Clore, G.M.; Omichinski, J.G.; Sakaguchi, K.; Zambrano, N.; Blessing, M.; Jorcano, J.; Balmain, A. Cell, 1990, 62, 697-708.
Sakamoto, H.; Appella, E.; Gronenborn, A.M. Science, 1995, 267 , [113] Yuspa, S.H.; Kilkenny, A.E.; Stanley, J.; Lichti, U. Nature, 1985,
1515-1516. 314, 459-462.
[77] Wang, X.W.; Vermeulen, W.; Coursen, J.D.; Gibson, M.; Lupold, [114] Albanese, C.; Johnson, J.; Watanabe, G.; Eklund, N.; Vu, D.;
S.E.; Forrester, K.; Xu, G.; Elmore, L.; Yeh, H.; Hoeijmakers, J.H. Arnold, A.; Pestell, R.G. J. Biol. Chem., 1995, 270, 23589-23597.
Genes Dev., 1996, 10, 1219-1232. [115] Medema, R.H.; Kops, G.J.; Bos, J.L.; Burgering, B.M. Nature,
[78] Kraiss, S.; Quaiser, A.; Oren, M.; Montenarh, M. J. Virol., 1988, 2000, 404, 782-787.
62, 4737-4744. [116] Chellappan, S.P.; Hiebert, S.; Mudryj, M.; Horowitz, J.M.; Nevins,
[79] Pietenpol, J.A.; Tokino, T.; Thiagalingam, S.; el-Deiry, W.S.; J.R. Cell, 1991, 65, 1053-1061.
Kinzler, K.W.; Vogelstein, B. Proc. Natl. Acad. Sci. U.S.A., 1994, [117] Winston, J.T.; Coats, S.R.; Wang, Y.Z.; Pledger, W.J. Oncogene,
91, 1998-2002. 1996, 12, 127-134.
[80] Slaton, J.W.; Benedict, W.F.; Dinney, C.P.N. Urology, 2001, 57, [118] Campisi, J. Eur. J. Cancer, 1997, 33, 703-709.
852-859. [119] Campisi, J. In Vivo, 2000, 14, 183-188.
 Anti-Cancer Drugs Mini-Reviews in Medicinal Chemistry, 2005, Vol. 5, No. 7 695

[120] Malumbres, M.; PerezDeCastro, I.; Hernandez, M.I.; Jimenez, [149] Tsai-Pflugfelder, M.; Liu, L.F.; Liu, A.A. Tewey, K.M.; Whang-
M.; Corral, T.; A. Mol. Cell Biol., 2000, 20, 2915-2925. Peng, J.; Knutsen, T.; Huebner, K.; Croce, C.M.; Wang, J.C. Proc.
[121] Ravi, R.K.; Weber, E.; McMahon, M.; Williams, J.R.; Baylin, S.; Natl. Acad. Sci. U.S.A., 1988, 85, 7177-7181.
Mal, A.; Harter, M.L.; Dillehay, L.E.: Claudio, P.P.; Giordano, A.; [150] Jenkins, J.R.; Ayton, P.; Jones, T.; Davies, S.L.; Simmons, D.L.;
Nelkin, B.D.; Mabry, M. J. Clin. Invest., 1998, 101, 153-159. Harris, A.L.; Sheer, D.; Hickson, I.D. Nucleic Acids Res., 1992,
[122] Nakanishi, H.; Brewer, K.A.; Exton, J.H. J. Biol. Chem., 1993, 20, 5587-5592.
268, 13-16. [151] Hwang, J.; Hwong, C.L. Adv. Pharmacol., 1994, 29A, 167-189.
[123] Braun, D.C.; Garfield, S.H.; Blumberg, P.M. J. Biol. Chem., 2004; [152] Woessner, R.D.; Mattern, M.R.; Mirabelli, C.K.; Johnson, R.K.,
[in Press as manuscript M413896200]. Drake, F.H., Cell Growth Diff., 1991, 2, 209-214.
[124] Wang, X.Y.; Repasky, E.; Liu, H.T. Exp. Cell. Res., 1999, 250, [153] Chen, A.Y., Liu, L.F. Ann. Rev. Pharmacol. Toxicol., 1994, 34,
253-263. 191- 218.
[125] Roychowdhury, D.; Lahn, M. Semin. Oncol., 2003, 30, 30-33. [154] Froelich-Ammon, S.J.; Osheroff, N. J. Biol. Chem., 1995, 270,
[126] Hofmann, J. Rev. Physiol. Biochem. Pharmacol., 2001, 142, 1-96. 21429-21432.
[127] Marshall, J.L.; Bangalore, N.; El-Ashry, D.; Fuxman, Y.; Johnson, [155] Pommier, Y.; Tanizawa, A.; Kohn, K.W. Adv. Pharmacol., 1994,
M.; Norris, B.; Oberst, M.; Ness, E.; Wojtowicz-Praga, S.; 29B, 73-92.
Bhargava, P.; Rizvi, N.; Baidas, S.; Hawkins, M.J. Cancer Biol. [156] Larsen, A.K.; Gobert, C. Pathol. Oncol. Res., 1999, 5, 171-178.
Ther., 2002, 1, 409-416. [157] Goto, T.; Laipis, P.; Wang, J.C. J. Biol. Chem., 1984, 259, 10422 -
[128] Swannie, H.C.; Kaye, S.B. Curr. Oncol. Rep., 2002, 4, 37-46. 10429.
[129] Liu, J.; Durrant, D.; Lee, R.M. Cancer Therapy, 2003, 1, 275-281. [158] Osheroff, N.; Zechiedrich, E.L.; Gale, K.C. BioEssays, 1991, 13,
[130] Cho-Chung Y. S.; Clair T. Pharmacol. Ther., 1996, 60, 265-288. 269-275.
[131] Cho-Chung Y. S.; Pepe S.; Clair T.; Budillon A.; Nesterova M. [159] Weinstein, J.N.; Myers, T.G; O’Connor, P.M.; Friend, S.H.;
Crit. Rev. Oncol. Hematol., 1995, 21, 33-61. Fornace, A.J.Jr.; Kohn, K.W.; Fojo, T.; Bates, S.E.; Rubinstein,
[132] Ciardiello F.; Tortora G. Clin. Cancer Res., 1998, 4, 821-828. L.V.; Anderson, N.L.; Buolamwini, J.K.; van Osdol, W.W.;
[133] Tortora G.; Ciardiello F. Ann. Oncol., 2000, 11, 777-783. Monks, A.P.; Scudiero, D.A.; Sausville, E.A.; Zaharevitz, D.W.;
[134] Carmena, M.; Earnshaw, W.C. Nat. Rev. Mol. Cell Biol., 2003, 4, Bunow, B.; Viswanadhan, V.N.; Johnson, G.S.; Wittes, R.E.; Paull,
842-854. K.D. Science, 1997, 275, 343-349.
[135] Nigg, E.A. Nat. Rev. Mol. Cell Biol., 2001, 2, 21-32. [160] Lowe, S.W.; Ruley, H.E.; Jacks, T.; Housman, D.E. Cell, 1993, 74,
[136] Bernard, M.; Sanseau, P.; Henry, C.; Couturier, A.; Prigent, C. 957-967.
Genomics, 1998, 53, 406-409. [161] Lowe, S.W.; Bodis, S.; McClatchey, A.; Remington, L.; Ruley,
[137] Kimura, M.; Matsuda, Y.; Yoshioka, T.; Okano, Y. J. Biol. Chem., H.E.; Fisher, D.E.; Housman, D.E.; Jacks, T. Science, 1994, 266 ,
1999, 274, 7334-7340. 807-810.
[138] Katayama, H.; Brinkley, W.R.; Sen, S. Cancer and Metastasis [162] Pai, H.H.; Rochon, L.; Clark, B.; Black, M.; Shenouda, G. Int. J.
Reviews, 2003, 22, 451-464. Radiat. Oncol. Biol. Phys., 1998, 41, 37-42.
[139] Vaux, D.L.; Cory, S.; Adams, J.M. Nature, 1988, 335, 440-442. [163] Wiggenraad, R.; Tamminga, R.; Blok, P.; Rouse, R.; Hermans, J.
[140] Melisi, D.; Troiani, T.; Damiano, V.; Tortora, G.; Ciardiello, F. Int. J. Radiat. Oncol. Biol. Phys., 1998, 41, 29-35.
Endocrine-Related Câncer, 2004, 11, 51-68. [164] Cote, R.J.; Esrig, D.; Groshen, S.; Jones, P.A.; Skinner, D.G.
[141] Adams, J.M.; Cory, S. Science, 1998, 281, 1322-1326. Nature, 1997, 385, 123-125.
[142] Juin, P.; Geneste, O.; Raimbaud, E.; Hickman, J.A. Biochim. [165] Tada, M.; Matsumoto, R.; Iggo, R.D.; Onimaru, R.; Shirato, H.;
Biophys. Acta, 2004, 1644, 251-260. Sawamura, Y.; Shinohe, Y. Cancer Res., 1998, 58, 1793-1797.
[143] Degterev, A.; Boyce, M.; Yuan, J. J. Cell Biol., 2001,155, 695- [166] Hickman, J.A. Eur. J. Cancer, 1996, 6, 921-926.
698. [167] Bellamy, W.T.; Dalton, W.S. Adv. Clin. Chem. 1994, 31, 1-61.
[144] Chan, S.L.; Lee, M.C.; Tan, K.O.; Yang, A.S.; Lee, A.S.; Flotow, [168] Shustik, C.; Dalton, W.; Gros, P. Mol. Aspects. Med. 1995, 16 , 1 -
H.; Fu, N.Y.; Buther, M.S.; Soerjato, D.D.; Bus, A.D.; Yu, V.C. J. 78.
Biol. Chem., 2003, 278, 20453-20456. [169] Chen, A.Y.; Liu, L.F. Adv. Pharmacol., 1994, 29B, 245-256.
[145] Beck, W.T.; Morgan, S.E.; Mo, Y.Y.; Bhat, U.G. Drug Resist. [170] Scheltema, J.M.; Romijn, J.C.; van-Steenbrugge, G.J.; Schroder,
Updat., 1999, 2, 382-389. F.H.; Mickisch, G.H. Anticancer Res., 2001, 5, 3161-3166.
[146] Cowell, I.G.; Okorokov, A.L.; Cutts, S.A.; Padget, K.; Bell, M.; [171] Granr, C.R.; Validjmarsson, G.; Hipfner, D.R.; Almquist, K.C.;
Milner, J.; Austin, C.A. Exp. Cell. Res., 2000, 255, 86-94. Cole, S.P.C.; Deelev, R.G. Cancer Res ., 1994, 54, 357-361.
[147] Beck, W.T.; Danks, M.K.; Wolverton, J.S. Adv. Pharmacol., 1994, [172] Shou, D.C.; Zittoun, R.; Marie, J.P. Leukemia , 1995, 9, 1661-1666.
145-169. [173] Friche, E.; Nissen, N.I.; Beck, W.T. Proc. Am. Assoc. Cancer
[148] Hochhauser, D.; Harris, A.L. Cancer Tret. Rev., 1993, 181-194. Res., 1995, 36, 217.

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