Control of vascular cell proliferation and migration by cyclin-dependent kinase
signalling: new perspectives and therapeutic potential
Vicente Andrés
Laboratory of Vascular Biology, Department of Molecular and Cellular Pathology and
Therapy, Instituto de Biomedicina de Valencia-CSIC, 46010 Valencia, Spain
WORD COUNT: 8021
CORRESPONDENCE TO:
Vicente Andrés
Instituto de Biomedicina de Valencia
C/Jaime Roig 11, 46010 Valencia (Spain)
Tel: +34-96-3391752
FAX: +34-96-3391750
E-mail: vandres@ibv.csic.es
1
ABSTRACT
Neointimal lesion development is a chronic inflammatory process that involves excessive
cell proliferation and migration within the artery wall. Progression through the mammalian cell
cycle requires the sequential activation of holoenzymes composed of a catalytic cyclindependent protein kinase (CDK) and a regulatory subunit named cyclin. Members of the CDK
family of inhibitory proteins (CKIs) interact with and inhibit the activity of CDKs. Cell
migration occurs predominantly at the G1/S phase of the cell cycle, and both CDKs and CKIs are
among the molecular machines that co-ordinately regulate the cycling events that control cell
proliferation and locomotion. The purpose of this review is to discuss the role of CDK/cyclin
holoenzymes and CKIs in the regulation of vascular cell proliferation and migration and in the
control of neointimal thickening. Pharmacological and gene therapy strategies targeting these
cell cycle regulators for the treatment of cardiovascular disease will be also discussed.
2
INTRODUCTION
The initiation and growth of atherosclerotic lesions is a complex multifactorial process that
involves adaptative and innate immune mechanisms [1-4]. Endothelial cell (EC) dysfunction
induced by atherogenic stimuli is one of the earliest manifestations of atherosclerosis at sites of
predisposition to atheroma formation (Fig. 1). The damaged endothelium promotes the adhesion
and transendothelial migration of circulating leukocytes. Early fatty streaks contain mostly highly
proliferative macrophages that avidly uptake lipoproteins to become lipid-laden foam cells.
Activated intimal leukocytes produce a plethora of inflammatory chemokines and cytokines that
promote the proliferation of vascular smooth muscle cell (VSMC) and their migration towards the
atherosclerotic lesion, thus further contributing to atheroma growth [1,2,5,6]. Excessive cell
proliferation and migration are also involved in the growth of vascular obstructive lesions during
restenosis post-angioplasty, transplant vasculopathy, and graft atherosclerosis. Plaque rupture or
erosion at advanced disease stages can lead to acute occlusion due to thrombus formation, resulting
in myocardial infarction or stroke.
While several proliferation markers are expressed in human primary atheroma and restenotic
lesions [7-16], the relevance of proliferation during human atherosclerosis and restenosis has been
controversial, with some studies reporting very low proliferative rates [8,9,11,13,15] and others
reporting abundance of dividing cells [10,17]. Cell proliferation appeared more pronounced in
restenotic versus primary lesions [11,17,18], and primary VSMCs obtained from human advanced
primary stenosing displayed diminished proliferation compared with cells from fresh restenosing
lesions [19], suggesting that cell proliferation is maximal at early stages of neointimal lesion
growth. Consistent with this interpretation, atheroma size and cellular proliferation within the
atheromatous plaque of hyperlipidemic rabbits are inversely correlated [20-22], and experimental
angioplasty is characterized by the reestablishment of the quiescent phenotype after the initial
proliferative burst [23,24].
3
The mammalian cell cycle is controlled by holoenzymes composed of a catalytic cyclindependent protein kinase (CDK) and a regulatory subunit named cyclin [25,26]. Diffferent
CDK/cyclin complexes are orderly activated at specific phases of the cell cycle (Fig. 2).
CDK/cyclin-dependent hyperphosphorylation of the retinoblastoma protein (pRb) and the related
pocket proteins p107 and p130 from mid G1 to mitosis contributes to the transactivation of genes
with functional E2F-binding sites, including several growth and cell-cycle regulators (i.e., c-myc,
pRb, cdc2, cyclin E, cyclin A), and genes encoding proteins required for nucleotide and DNA
biosynthesis (i. e., DNA polymerase , histone H2A, proliferating cell nuclear antigen, thymidine
kinase) [27-30]. The identities of substrates of the yeast CDK1 (CDC2) have revealed that this
enzyme employs a global regulatory strategy involving phosphorylation of other regulatory
molecules as well as phosphorylation of the molecular machines that drive cell-cycle events [31].
Cyclin availability and phospo/dephosphorylation of CDKs and cyclins by specific kinases
and phosphatases regulate the activity of CDK/cyclin holoenzymes. Of central importance in cell
cycle regulation, CDK activity is attenuated by the interaction with CDK inhibitory proteins
(CKIs) of the Cip/Kip (for CDK interacting protein/Kinase inhibitory protein) and Ink4 (for
inhibitor of CDK4) families [32] (Fig. 2). Cip/Kip proteins (p21Cip1, p27Kip1, p57Kip2) bind to and
inhibit a wide spectrum of CDK/cyclin holoenzymes, while Ink4 proteins (p16 Ink4a, p15Ink4b,
p18Ink4c, p19Ink4d) are specific for cyclin D-associated CDKs. Mitogenic and antimitogenic stimuli
affect the rates of CKI synthesis and degradation, as well as their redistribution among different
CDK/cyclin heterodimers.
Control of vascular cell proliferation and neointimal lesion growth by CDKs and CKIs
VSMC proliferation in the balloon-injured rat carotid artery is associated with a temporally
and spatially coordinated expression of CDKs and cyclins [16,33], and augmented expression of
these proteins is associated with an increase in their kinase activity [16,34]. CDK and cyclin
4
expression has been also detected in human VSMCs within atherosclerotic and restenotic tissue
[10,16,35]. Collectively, these findings suggest that the assembly of functional CDK/cyclin
holoenzymes in the injured arterial wall is a hallmark of vascular proliferative disease.
p27Kip1 and p21Cip1 have been implicated in the mechanism of action of several
pharmacological agents that control vascular cell proliferation in vitro and neointimal thickening.
Treatment of VSMCs with salicylate prevented PDGF-induced downregulation of p27Kip1 and
p21Cip1 but not of p16Ink4a, and this was accompanied by reduced CDK2 activity and growth
arrest [36]. Likewise, beraprost sodium-dependent VSMC growth arrest and reduction of intimal
thickening in the balloon-injured dog coronary artery correlated with maintained p27Kip1
expression [37]. Upregulation of p27Kip1 and p21Cip1 may be one mechanism by which nonsteroidal anti-inflammatory drugs [38], nitric oxide donors [39] and gene transfer of endothelial
nitric oxide synthase induce VSMC growth arrest [40]. Similarly, induction of p21Cip1 was
associated with tranilast-dependent inhibition of CDK2 and CDK4 activities, VSMC growth
arrest in vitro and reduced intimal hyperplasia in the rat balloon-injured carotid artery [41].
Prevention of Rho GTPase-induced downregulation of p27Kip1 without changes in p21Cip1,
p16Ink4a, or p53 levels may mediate simvastatin-dependent inhibition of CDK2 activity and
VSMC proliferation [42]. Otterbein et al. have recently shown that exposure of VSMC cultures
to carbon monoxide (CO) transiently increases p21Cip1 expression and results in growth arrest
[43]. Notably, CO suppressed arteriosclerotic lesions associated with both chronic graft rejection
and with balloon injury in rats. However, although p21Cip1 was essential for CO-dependent
VSMC growth arrest in vitro, the therapeutic effect of CO in a mouse model of mechanical
arterial injury was not impaired in p21Cip1-null mice [43].
Hemodynamic forces are thought to play an important role in the initiation and
progression of atherosclerotic lesions [1,2]. Steady laminar stress induced EC growth arrest and
this correlated with p21Cip1 upregulation without changes in p27Kip1 protein levels [44]. On the
5
other hand, stretch-mediated inactivation of forkhead transcription factors and p27Kip1
downregulation in VSMCs was accompanied by activation of CDK2, pRb hyperphosphorylation
and proliferation, demonstrating that the earliest cell cycle events in VSMCs can occur in a
solely mechanosensitive fashion [45]. RhoA-dependent reduction of p27Kip1 expression,
mediated in part via phosphatidylinositol-3-kinase, induces VSMC proliferation and may
contribute to the enhanced vascular responsiveness associated to hypertension [46].
Cell cycle progression in the artery wall is regulated by specific components of the
extracellular matrix (ECM) and integrins [47]. Neointimal VSMCs synthesize novel ECM
components and induce the expression of matrix-degrading proteases that remodel the
surrounding ECM. Notably, matrix-degrading metalloproteinase expression is induced within
neointimal lesions [48-51], and metalloproteinase inhibitors repressed VSMC proliferation in
vitro and after angioplasty [52-54]. Significant changes in collagen content occur during
neointimal lesion development [55-57]. Because polymerized collagen may mimic the scenario
of a normal artery composed of quiescent VSMCs, and monomer collagen might resemble the
ECM surrounding proliferating VSMCs within atherosclerotic and restenotic plaques, Koyama et
al. studied the growth properties of VSMCs cultured on monomer collagen fibers and on
polymerized collagen [58]. Mitogen-stimulated VSMCs grown on monomer collagen disclosed
high proliferative activity, but underwent G1 arrest when seeded on polymerized collagen. This
inhibitory effect of polymerized collagen appeared to be mediated by 2 integrins, and correlated
with suppression of p70S6K and upregulation of p27Kip1 (and to a lesser extent p21Cip1). Thus,
regulation of CKIs in response to changes in specific ECM components might regulate the ability
of VSMCs to respond to growth signals in vitro. Interestingly, the quiescent phenotype of
nonadherent NRK fibroblasts correlated with an increased association of p27Kip1 and p21Cip1 to
cyclin E-containing holoenzymes [59]. Further studies are required to determine whether
6
changes in arterial CKI expression regulate cell proliferation in response to integrins and ECM
components in vivo.
The gradual increase in p21Cip1 and p27Kip1 observed in balloon-injured rat and porcine
arteries suggests that these factors may limit neointimal hyperplasia after the initial proliferative
wave [14,60,61]. Using a mouse model of transluminal femoral artery injury, Reis et al. showed
a rapid apoptotic response and downregulation of p27Kip1 in medial VSMCs, which was followed
by a gradual increase in cell proliferation that peaked at 2 weeks in both the media and neointima
and decreased thereafter [62]. Restoration of low proliferative activity during later phases of
vascular repair in this model correlated with increased p27Kip1 expression. Several studies have
suggested a role for CKIs in the regulation of cell proliferation during human neointimal
thickening: a) reduced p27Kip1expression was detected in primary atherosclerotic lesions
compared with that in aorta, internal mammary artery, and carotid artery thrombendarterectomy
specimens, and in lesions of in-stent restenosis patients [63]. These authors also found a
significant upregulation of p21Cip1 in estenosis compared with primary lesions and other vascular
regions; b) more frequent expression of p27Kip1 and p21Cip1 was found within regions of human
coronary atheromas not undergoing proliferation [14]; c) concordant expression of TGF-
receptors I and II in virtually all cells positive for p27Kip1 within human atherosclerotic plaques
suggests that the anti-mitogenic action of TGF-1 in these lesions may be mediated by p27Kip1
[35]; d) coexpression of p53 and p21Cip1 in human carotid atheromatous plaque cells that
revealed lack of proliferation markers suggests that induction of p21Cip1 may occur via p53dependent transcriptional activation [64]; e) attenuation of PDFGF-BB-induced p21Cip1 and
p27Kip1 expression by interleukin-1 may promote VSMC hyperplastic growth after vascular
injury and in atherosclerosis [65].
Collectively, the above studies suggest an important role of p21Cip1 and p27Kip1 in
neointimal lesion growth. We have established a causal link between decreased p27Kip1 protein
7
expression and atherogenesis in hypercholesterolemic apolipoprotein E (apoE)-null mice by
demonstrating that whole-body genetic inactivation of p27Kip1 increases arterial VSMC and
macrophage proliferation and accelerates atherosclerosis [66]. In another study, we have shown
that selective inactivation of p27Kip1 in hematopoietic progenitor cells increases neointimal
macrophage proliferation and accelerates atherosclerosis in fat-fed apoE-deficient mice [67],
consistent with previous studies demonstrating enhanced haematopoietic progenitor cell
proliferation upon p27Kip1 inactivation [68], and implicating p27Kip1 as a critical macrophage
growth suppressor [69,70]. Because macrophages were the most abundant neointimal cells in our
study [67], it seems reasonable to suggest that macrophage p27Kip1 safeguards against the
inflammatory/proliferative response induced by dietary cholesterol in apoE-null mice. Regarding
the consequences of CKI inactivation on neointimal thickening induced by mechanical injury,
Otterbein et al. reported 3 times more pronounced lesion size in p21Cip1-null versus wild-type
mice [43]. In contrast, neointimal hyperplasia after mechanical vascular damage was similar in
wild-type and p27Kip1-null mice [71]. Redundant roles between p21Cip1 and p27Kip1, or a
compensatory increase in p21Cip1 expression (or other CKIs) might account for the lack of
phenotype of p27Kip1-null mice in the setting of mechanical arterial injury.
Animal and human studies have recognized significant differences in the atherogenicity of
different segments of the arterial, which may be related to regional phenotypic variance of
VSMCs, both when comparing cells from different compartments of the same vessel or cells
isolated from vessels from different vascular beds [72-78]. Sustained p27Kip1 expression in spite
of growth stimuli may contribute to the resistance to growth of human VSMCs isolated from
internal mammary artery compared with saphenous vein VSMCs, and to the longer patency of
arterial versus venous grafts [77]. Likewise, distinct p15Ink4b and p27Kip1 expression correlated
with different proliferative potential of intimal and medial VSMCs stimulated with basic
fibroblast growth factor [78], and intrinsic regional differences in the proliferative and migratory
8
capacity of VSMCs due to distinct regulation of p27Kip1 may contribute to creating variability in
atherogenicity in different vascular beds [79].
CDK inhibitory approaches to reduce neointimal thickening
The importance of CDK activation for neointimal lesion growth has been demonstrated by
means of pharmacological (Table 1) and gene therapy (Table 2) CDK inhibitory strategies. CVT313 is a purine derivative that inhibits CDK activity by preventing the binding of ATP to the
adenine-binding pocket of CDKs [80-82]. The relative inhibitory potency of CVT-313 varies
from very high for CDK2, moderate for CDK1, and very low for CDK4 [83]. Flavopiridol (L868275) is a more potent CDK inhibitor that displays higher specificity towards CDK4 than
towards CDK1 and CDK2 [84,85]. Growth arrest in VSMCs treated with flavopiridol and CVT313 correlates with decreased pRb protein levels and/or inhibition of its phosphorylation [83,86].
It is noteworthy that CVT-313 [83] and flavopiridol [85,87] can cause blockade in different cell
cycle phases depending on both the concentration of the drug and the cell line analyzed. In the rat
carotid model of balloon angioplasty, a brief intraluminal exposure of CVT-313 [83] or oral
administration of flavopiridol for 5 days beginning at the day of injury [86] reduced neointima
formation by 80% and 39%, respectively.
Gene therapy approaches based on the use of antisense oligodeoxynucleotide (ODN)
targeting CDKs and cyclins have shown efficacy in reducing neointimal lesion formation in
animal models of balloon angioplasty, including ODN against CDK2 [34,88], CDC2 [34,89,90],
and cyclin B1 [90]. Likewise, downregulation of cyclin G1 expression by retrovirus-mediated
antisense gene transfer inhibited VSMC proliferation and neointima formation after balloon
angioplasty [91]. Antisense ODN to CDC2/PCNA [92] and CDK2 [93] also attenuated graft
atherosclerosis. Additional approaches based on the inactivation of positive cell cycle regulators
that do not directly target CDK/cyclin activity (i. e., E2F, c-myc, etc) have been reviewed
9
elsewhere [6,94].
Consistent with the notion that CKIs function as negative regulators of neointimal
thickening (see above), gene transfer of p21Cip1 [95-97], p27Kip1 [60,98], and p57Kip2 [99] reduced
neointima formation after angioplasty in normocholesterolemic animals. Likewise, p21Cip1
overexpression attenuated neointimal thickening after balloon injury in hypercholesterolemic
mice [100] and following vein grafting [101]. On the other hand, antisense ODN to p21Cip1
attenuated matrix protein secretion in VSMCs [102]. Lamphere et al. generated chimeric p16Ink4a
and p27Kip1 molecules, which were of comparable potency to the parental p27Kip1 in inhibiting the
activities of several CDKs in vitro [103]. Among these chimeras, W9 was the most potent
growth suppressor of human coronary artery VSMCs and ECs when compared to the parental
p16Ink4a and p27Kip1, p27Kip1 derivatives, or several alternative p27Kip1-p16Ink4a chimeras.
Moreover, W9 was more effective in inhibiting neointimal thickening after balloon angioplasty
in cholesterol-fed rabbits [104]. Thus, combining the activities of different CKIs might increase
the therapeutical activity in the treatment of neointimal thickening after angioplasty. Further
approaches based on the overexpression of growth suppressor that do not directly target
CDK/cyclin activity (i. e., pRb, p53, Gax, etc) are reviewed elsewhere [6,94].
Control of cell migration by CDKs and CKIs
Several cytostatic agents (eg, quercetin, mimosine, suramin, rapamycin, and troglitazone)
can reduce the migratory potential of VSMCs and tumor cells [105-110]. Likewise, 17-estradiol
and the transcription factors p53, AP-1 and c-myc regulate in a coordinated manner the
proliferative and migratory potential of ECs and VSMCs [111-114]. NBT-II rat bladder
carcinoma cells synchronized in G1 migrated simultaneously upon FGF-1 stimulation, and cells
arrested in G2/M did not respond to stimulation by this mitogen [115]. Moreover, maximal
migration of PDGF-BB-stimulated VSMCs occurred in late G1 [116]. These studies indicate that
10
the position in the cell cycle is a key determinant of a cell’s competence for migration. Of note in
this regard, diverse cytoskeletal reorganization genes and many genes involved in cell motility
and remodeling of the extracellular matrix exhibited cell-cycle dependent regulation in human
fibroblasts [117].
Overexpression of p27Kip1, p16INK4a and p21Cip1 inhibits cell spreading and migration in
human umbilical vein ECs, CS-1 3 melanoma cells, VSMCs and NIH-3T3 fibroblasts [79,118120]. Moreover, p27Kip1-null VSMCs were more resistant than wild-type cells to the
antimigratory properties of rapamycin [121]. Regarding the role of CDKs on cell locomotion,
CDK6 localized to the ruffling edge of spreading cells and suppressed p16INK4a-mediated
inhibition of cell spreading [119]. Likewise, CDK5 activity has been involved in the regulation
of specific components of neuronal migration at different developmental stages [122], as well as
in the modulation of actin cytoskeleton dynamics in cells [123]. Moreover, CDK5 plays a key
role in regulating morphology, cell adhesion, and apoptosis in the human astrocytoma cell line
U373 [124]
In view of the above connections between cell proliferation and migration, we
investigated whether the dual function of p27Kip1 as a cell-cycle and migration inhibitor is
achieved via common or independent molecular pathways [125]. We found that physiologically
high level of p27Kip1 expression inhibits CDK activity and attenuates both proliferation and
migration in VSMC and fibroblast cultures. Mutations that rendered p27Kip1 unable to abrogate
CDK activity also prevented p27Kip1-induced growth arrest and migration blockade. We also
showed that a constitutively active mutant of pRb insensitive to CDK-dependent
hyperphosphorylation inhibited both cell proliferation and migration. In contrast, pRb
inactivation by forced expression of the adenoviral oncogene E1A correlated with high
proliferative and migratory activity. Collectively, these results suggest that cellular proliferation
and migration are regulated in a coordinated manner by the p27 Kip1/CDK/pRb/E2F pathway (Fig.
11
3). Consistent with this notion, E2F-1-null keratinocytes exhibited delay in transit through both
G1 and S phases of the cell cycle and substantially impaired migration [126]. Future studies are
necessary to identify E2F-regulated genes implicated in cell locomotion.
Concluding remarks
Excessive cell proliferation and migration contribute to neointimal thickening. It has been
well established that CDKs, cyclins and CKIs are key regulators of these processes in vitro.
Moreover, changes in the expression and/or activity of these cell cycle regulators have been
documented in several animal models of vascular proliferative disease and in human
atherosclerotic and restenotic tissue. Importantly, synthetic CDK inhibitors (CVT-313,
flavopiridol), antisense ODN to CDK/cyclins, and CKI overexpression reduced neointimal
hyperplasia in the setting of experimental graft atherosclerosis and angioplasty. Although these
CDK inhibitory strategies have not been assessed in clinic, antiproliferative approaches that have
shown promising results for preventing human neointimal thickening are available [6,94]. The
bacterial macrolide rapamycin (sirolimus, rapamune) is the pharmacological agent with which
most experience has been gathered so far for the prevention of in-stent restenosis. Rapamycin is
a potent immunosuppressant that strongly inhibits VSMC proliferation and migration
[105,106,121,127] via both p27Kip1–dependent [121,127] and p27Kip1–independent [33,71]
mechanisms. Rapamycin potently inhibited neointimal thickening in animal models of
angioplasty, graft atherosclerosis, and diet-induced atherosclerosis [71,128-136], and recent
clinical trials using rapamycin-impregnated stents have shown promising results for the
prevention of neointimal proliferation, restenosis, and associated clinical events in patients
undergoing coronary angioplasty [137-139]. Activation of E2F is triggered by CDK-dependent
phosphorylation of pRb and pocket proteins, therefore blockade of E2F function may be a
common mechanism by which different CDK/cyclin inhibitory strategies reduce neointimal
12
thickening. E2F inactivation via transfection of synthetic ODN containing an E2F consensus
binding site reduced experimental hyperplasia after balloon angioplasty and vein grafting
[140,141], and application of this E2F ‘decoy’ strategy is safe and can achieve sequence-specific
inhibition of cell-cycle gene expression and DNA replication in patients receiving bypass vein
grafts [142,143]. Despite these encouraging results, significant effort in basic research is
warranted to identify additional target genes and strategies for the treatment of cardiovascular
disease.
ACKNOWLEDGEMENTS
I apologize to colleagues whose work has not been directly cited due to space limitations. I
thank María J. Andrés-Manzano for preparing the figures. Work in my laboratory is currently
supported by grants from the Spanish Ministry of Science and Technology and Fondo Europeo de
Desarrollo Regional (SAF2001-2358, SAF2002-1443), and from Instituto de Salud Carlos III
(Red de Centros C03/01).
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Figure 1: Atheroma development is a multifactorial process. Endothelial dysfunction induced
by different atherogenic stimuli triggers a chronic inflammatory response within the artery wall
that results in excessive proliferation and migration of leukocytes and VSMCs. At advanced
disease states, plaque rupture or erosion can lead to thrombus formation and associated ischemic
events.
Figure 2: Cell cycle control in mammalian cells. Activation of specific CDK/cyclin complexes
drives progression through the cell cycle (CDK1=CDC2). CKIs interact with and inactivate
CDK/cyclin holoenzymes.
Figure 3: Coordinate control of cell proliferation and migration. In the presence of low level
of p27Kip1 protein, active CDK/cyclin holoenzymes trigger the hyperphosphorylation of pRb,
release of E2F and high proliferative and migratory activity. In contrast, CDK/cyclin inactivation
by high level of p27Kip1 leads to the accumulation of hypophosphorylated pRb, sequestration of
E2F and low proliferative and migratory activity [79,120,125]. High p21Cip1 protein level also
induces growth arrest and migration blockade [118].
14
Table 1. Pharmacological CDK inhibitors that limit neointimal hyperplasia in the rat
carotid artery model of balloon angioplasty
CDK inhibitor
IC50 for CDKs
CDK1 (CDC2)= 4 M
CVT-313
(Purine derivative) CDK2=0.5 M
Dose and route of administration
1.25 mg/kg, brief intraluminal
exposure immediately after
angioplasty
Ref.
[83]
CDK4=215 M
CDK1 (CDC2)=0.5 M
Flavopiridol
CDK2=0.1 M
(Flavonoid)
CDK4=0.065 M
5 mg/kg/day, oral administration
CDK6=0.06 M
CDK7=0.11-0.3 M
15
[84-86,144]
Table 2. Gene therapy strategies targeting CDK/cyclin activity with beneficial effects in
animal models of cardiovascular disease
Strategy
Antisense-mediated
CDK/cyclin inactivation
Targeted gene
Strategy
Animal model
CDK2
ODN
Balloon angioplasty (rat)
CDK2
ODN
Graft atherosclerosis (mouse) [93]
CDC2
ODN
Balloon angioplasty (rat)
[34,89,90]
cyclin B1
ODN
Balloon angioplasty (rat)
[90]
CDC2/PCNA
ODN
Graft atherosclerosis
(rabbit, rat)
[92]
Balloon angioplasty (rat)
[91]
cyclin G1
Retrovirus
p21Cip1
Adenovirus Balloon angioplasty
(rat, mouse, pig)
p21Cip1
Plasmid
Ref.
[34,88]
[95-98,100]
Graft atherosclerosis (rabbit) [101]
CKI overexpression
p27Kip1
Adenovirus Balloon angioplasty
(rat, pig)
p57Kip2
Adenovirus Balloon angioplasty (rabbit)
[60,98]
[99]
p27Kip1-p16Ink4a Adenovirus Balloon angioplasty (rabbit) [104]
chimera
16
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