VEGF and Receptors
Pradeep
10.1177/1534735405282557
VEGF and
et Receptors
al
Expression of Vascular Endothelial
Growth Factor (VEGF) and VEGF Receptors
in Tumor Angiogenesis and Malignancies
C. R. Pradeep, MSc, E. S. Sunila, MSc, and G. Kuttan, PhD
Angiogenesis is a process by which new blood vessels are
formed from preexisting vessels. New blood vessel forma-
tion by angiogenesis involves the degradation of extra-
cellular matrix combined with sprouting and migration of
endothelial cells from preexisting capillaries. Solid tumors
consist of several components, including normal and
stromal cells, extracellular matrix, and vasculature. To grow
and metastasize, tumors must stimulate the development of
new vasculature through angiogenesis. Vascular endothelial
growth factor (VEGF) is a potent angiogenic peptide with bi-
ologic effects that include regulation of hematopoietic stem
cell development, extracellular matrix remodeling, and in-
flammatory cytokine regeneration. VEGF is both a vascular
growth factor and a vascular permeability factor. Its expres-
sion can upregulate several proangiogenic and prometa-
static molecules. As a central mediator of angiogenesis,
VEGF has emerged as an important target for antiangio-
genic therapy. In this review, the authors describe the essen-
tial characteristics of VEGF and the VEGF family of ligands
and their receptors. They also provide an overview of the
central role of VEGF in physiologic and pathologic angio-
genesis, directly or indirectly. This review sheds light on the
importance of VEGF-targeted antiangiogenic therapy based
on the monoclonal antibodies against VEGF, small inter-
fering RNA, and therapy directed against VEGF-VEGFR
kinase. It also gives a brief overview of the natural products
or dietary compounds that could be used as antiangiogenic
agents. Therapeutic inhibition of vessel formation could be
best suited to preventive strategies aimed at the suppression
of angiogenesis in primary tumors in subjects at risk or of
micrometastases after surgical removal of primary tumor.
Keywords: angiogenesis; VEGF; HIF-1 ; endothelial cells
The development of vascular supply is a fundamental
requirement for organ development and differentia-
tion during embryogenesis as well as for wound heal-
ing and reproductive functions in adults. Angio-
genesis is also indicated in the pathogenesis of a
variety of disorders: proliferative retinopathies, age-
related macular degeneration, tumors, rheumatoid
DOI: 10.1177/1534735405282557
INTEGRATIVE CANCER THERAPIES 4(4); 2005 pp. 315-321
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1,2
arthritis, and psoriasis. Vascular endothelial growth
factor (VEGF) is the only growth factor proven to be
specific and critical as a mitogen for vascular endothe-
lial cells derived from arteries, veins, and lymphatics,
but it is devoid of consistent and appreciable mito-
genic activity for other cell types.3 VEGF promotes
angiogenesis in tridimensional in vitro models, induc-
ing confluent microvascular endothelial cells to in-
vade collagen gels and form capillary-like structures.4
Even though VEGF is a potent mitogenic stimulator of
endothelial cells, it may also exert effects on cells in a
manner unrelated to its angiogenic activity and act as a
survival factor. An inverse relationship exists between
apoptosis and angiogenesis,5,6 several studies have
demonstrated the ability of VEGF to function as a
survival factor for endothelial cells. Immature blood
vessels such as those present in tumors with active
angiogenesis are dependent on the presence of VEGF,
7
undergoing regression via apoptosis.
VEGF induces vasodilatation in vitro and produces
a transient tachycardia, hypotension, and decrease in
8
cardiac output. VEGF is also known as a vascular leak-
age promoter in guinea pig. It is a potent angiogenic
peptide with diverse biological activities that include
regulation of embryonic stem cell development, ex-
tracellular matrix remodeling, and the local genera-
tion of inflammatory cytokines.9 Several excellent re-
views of the VEGF family have been published, and the
10-12
reader is referred to these for more details.
VEGF and Receptors
VEGF is a family of closely related growth factors hav-
ing a conserved pattern of 8 cysteine residues and
sharing common VEGF receptors (Figure 1). Vascu-
lar endothelial growth factor is commonly known as
CRP is at the Department of Biotherapeutics, Avestha Gengrain
Technologies Pvt Ltd, Bangalore, India. ESS and GK are at the
Department of Immunology, Amala Cancer Research Centre,
Thrissur-Kerala, India.
Correspondence: G. Kuttan, Department of Immunology, Amala
Cancer Research Centre, Thrissur-Kerala, India 680 555.
315
 Pradeep et al

Figure 1 Vascular endothelial growth factor (VEGF) receptor and ligands. The receptors for VEGF and related ligands include VEGFR-1,
VEGFR-2, and VEGFR-3. These receptors have multiple immunoglobulin G–like extracellular domain and intracellular tyrosine
kinase activity.

VEGF-A. Four additional members of the family such
as placental growth factor (PIGF), VEGF-B, VEGF-C,
and VEGF-D have been identified to date. VEGF is a
potent growth factor for blood vessel endothelial cells,
showing pleiotropic responses that facilitate migra-
tion, proliferation, tube formation, and survival of en-
dothelial cells; thus, VEGF is a common link of inflam-
mation, permeability, and angiogenesis.13,14
VEGF-A may exist in 4 isoforms, designated by their
expected final amino acid length (VEGF121, VEGF165,
VEGF189, VEGF206). These isoforms show similar biolog-
ical activities but bind with different affinities to hepa-
rin and result in different secretion patterns. VEGF121
is the smallest isoform of VEGF, stabilized by intra- and
interdisulfide bonds. VEGF121 is secreted and com-
pletely diffusible, but the largest isoform, VEGF206, is
almost completely attached to the extracellular ma-
trix. The other 2 show intermediate heparin-binding
affinities. VEGF-A exerts its action through 2 recep-
13,14
tors, VEGFR-1 and VEGFR-2. VEGFR-1 appears to
function primarily as a decoy receptor, which de-
creases the availability of VEGF to VEGFR-2.
PIGF is expressed in the placenta and somewhat
less in the heart, lung, and thyroid gland. Placentally
expressed PIGF may act as an autocrine growth factor
on trophoblasts, which express both PIGF and its re-
ceptor (VEGFR-1). Two differentially spliced isoforms
of PIGF have been identified. Hypoxia does not induce
PIGF synthesis, but the formation of heterodimers
would be affected due to hypoxic control over VEGF-A
expression. PIGF exerts its action through the recep-
13,15,16
tor VEGFR-1.
VEGF-B is largely cell associated and expressed
mostly in the heart, skeletal muscle, brain, and kidney.
It is often coexpressed with VEGF-A, and heterodi-
mers of A/B have been identified. VEGF-B expression
is not regulated by hypoxia. The long half-life of its
mRNA (>8 hours) suggests a chronic rather than acute

316 INTEGRATIVE CANCER THERAPIES 4(4); 2005

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 VEGF and Receptors

Figure 2 Effect of vascular endothelial growth factor (VEGF) on the microvessel outgrowth of rat aortic ring. The thoracic aorta was
removed from Wistar rat. Using a dissecting microscope, 1-mm-long rings were cut. Each ring was rinsed 8 times with culture
medium containing 100 U/mL penicillin and 100 g/mL streptomycin in the presence and absence of VEGF (20 g/mL). The
aortic preparations were cultured for 6 days, and microvessel growth was assessed daily (figure is from the sixth day). (a) Rat
thoracic aorta in the presence of VEGF (20 g/mL). (b) Rat thoracic aorta in the absence of VEGF (20 g/mL).

regulation. VEGF-A exerts its action through one
15,17-19
receptor, VEGFR-1.
VEGF-C, also called VEGF regulated factor or
VEGF-2, in the adult is expressed primarily in the
heart, placenta, lung, kidney, muscle, ovary, and small
intestine. During embryonic development, it is ex-
pressed in the cephalic mesenchyme, tail region,
allantois, and along the somites. VEGF-C exerts its
action through two receptors, VEGFR-2 and VEGFR-3.20
VEGF-D, also called c-fos induced growth factor, is a
VEGF homologue induced by c-fos. It is expressed in
adult lung, heart, and small intestine and in the fetal
lung and reported to be mildly mitogenic for endothe-
lial cells. VEGF-D and VEGF-C share 23% amino
sequence homology. VEGF-D exerts its action through
two receptors, VEGFR-1 and VEGFR-2.21-23
Hypoxia Inducible Factor-1
and VEGF Expression
The mechanism proposed to participate in the regula-
tion of VEGF gene expression, oxygen tension, plays a
24
major role both in in vitro and in vivo systems. VEGF
mRNA expression is rapidly and reversibly induced by
exposure to low pO2 in a variety of normal and trans-
25
formed cultured cell types. Ischemia with occlusion
in the artery results in a dramatic increase in VEGF
mRNA levels in in vivo experimental models, suggest-
ing the possibility that VEGF may mediate the spon-
taneous revascularization that follows ischemia.26
Hypoxia-induced transcription of VEGF mRNA is ap-
parently mediated, at least in part, by the binding of
the hypoxia-inducible factor-1 (HIF-1) to an HIF-1
27
binding site located in the VEGF promoter. HIF-1 has
been identified as a mediator of transcriptional re-
sponse to hypoxia and is a basic, heterodimeric, helix-
loop-helix protein.27 It turns out that some mecha-
nisms that lead to elevated VEGF production inde-
pendently of hypoxia actually short-circuit the normal
hypoxia-sensing mechanism that regulates VEGF ex-
pression. For example, it was observed that the onco-
gene v-src can induce expression of HIF-1, thereby
short circuiting the HIF-1-dependent hypoxia-sensing
mechanism and leading to increased expression of
VEGF.28 In addition to the induction of transcription,
hypoxia promotes stabilization of the VEGF mRNA by
proteins that bind to sequences located in the 3′ un-
translated region of the VEGF mRNA. Recently, one
such protein has been identified as the HuR mRNA
binding protein.29 It is clear that additional proteins
stabilize the VEGF mRNA, but their identity is still
unknown.
VEGF and Physiological Angiogenesis
The proliferation of blood vessels is crucial for a wide
variety of physiological processes such as embryonic
development, normal growth and differentiation,
wound healing, and reproductive functions. During
embryogenesis, blood vessels form through 2 distinct
processes, vasculogenesis and angiogenesis. Disrup-
tions of both VEGF alleles in mice mimics knockout of
VEGFR-2, resulting in almost complete absence of a
vasculature.30 Disruption of even a single VEGF allele
in mice leads to embryonic lethality due to severe vas-
31
cular abnormalities. The most elegant demonstra-

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 Pradeep et al
Figure 3 Influence of vascular endothelial growth factor (VEGF) on human umbilical vein endothelial cells (HUVECs) motility. HUVECs
were grown to confluence on a gelatin precoated 96-well titer plate. When the cultures reached 70% confluency (2-3 days), a
zone of cells was scraped away with a sterile razor blade and fresh 199 medium was added containing 100 U/mL penicillin and
100 g/mL streptomycin in the presence and absence of VEGF for 48 hours. (a) HUVEC in the presence of VEGF (20 ηg/mL).
(b) HUVEC in the absence of VEGF (20 g/mL).
tion of the need for exquisite VEGF regulation in-
volves retinal vascularization, which occurs postnatally
in rodents. Angiogenic sprouting into the initially
avascular and hypoxic rodent retina depends on its
VEGF expression.32 Any perturbation of normal VEGF
expression patterns destroys retinal vascularization
33
patterns with dire results for retinal function.
Spontaneous angiogenic response of VEGF can be
demonstrated using rat thoracic aortic culture. The
complexity of the vascular network could be observed
in the rat aortic preparations that were cultured in the
presence of VEGF (Figure 2). A complex network of
branching microvessels develops from the endothelial
cells of the aortic intima, interspersed with fibroblasts
as individual cells, thus providing a closer approxima-
tion to the angiogenic process as observed in vivo. This
process occurs in response to endogenous growth fac-
tors such as VEGF in the controlled microenviron-
ment. Angiogenesis is a complex morphogenic pro-
cess involving the coordinated migration of several
cell types including endothelial cells, peritypes, and
stromal fibroblasts. Figure 3 demonstrates the impact
of VEGF on endothelial cell migration or motility.
VEGF and Tumor-Specific Angiogenesis
Neovascularization is associated with numerous path-
ologic processes including cancer, proliferative dia-
betic retinopathy, and macular degeneration. The
pathological angiogenesis necessary for active tumor
growth is sustained and persistent with the initial ac-
quisition of the angiogenic phenotype being a com-
mon mechanism for the development of a variety of
solid and hematopoetic tumor types.34
Among the angiogenic growth factors, the VEGF
family (VEGF-A, VEGF-B, VEGF-C, and VEGF-D) and
318
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their corresponding receptor tyrosine kinases (VEGFR-
1 [FLT-1], VEGFR-2 [FLK-1, KDR], and VEGFR-3
[FLT-4]) play a paramount and indispensable role
in regulating the multiple facets of the angiogenic
and lymphangiogenic processes (VEGFR-3 [FLT-4])
as well as the induction of vascular permeability and
35
inflammation.
The VEGF family has a relatively narrow target cell
specificity compared to other pleiotropic angiogenic
factors and cytokines, which exert their mitogenic,
chemotactic, and thrombogenic effects primarily on
endothelial cells implicated in hemangiogenesis
(VEGF-A, VEGF-B) and lymphangiogenesis (VEGF-C
and VEGF-D). The endotheliotropic activities of the
VEGF family are mediated through the receptor tyro-
sine kinases VEGFR-1/FLT-1, VEGFR-2/KDR, and
VEGFR-3/FLT-4, the expression of which is upregu-
lated on vascular endothelial cells during embryonic
and tumor angiogenesis.
36
The receptor VEGF-R2/KDR is the principal one
through which VEGFs exert their mitogenic, chemo-
tactic, and vascular permeability effects on the host
37
vasculature. Increased expression of VEGFs by
tumor-associated vasculature is a hallmark of a vari-
ety of human and rodent tumors in vivo and is cor-
related with tumor growth rate, microvessel density,
proliferation, tumor metastatic potential, and poorer
38
patient prognosis in a variety of malignancies. The
collective findings show the critical and nonredun-
dant role of VEGF in tumor-associated angiogenesis,
mediated by chemotactic activity in monocytes and
39
macrophages, regulating extracellular matrix pro-
40
teolytic activity and release of tissue factor and nitric
39
oxide in endothelial cells and trophoblasts, and
mediating PIGF-induced recruitment and mobiliza-
INTEGRATIVE CANCER THERAPIES 4(4); 2005

tion of bone marrow–derived endothelial and hema-
topoetic stem cells in normal and tumor-associated
41
angiogenesis. Much evidence implicates VEGFR-3/
FLT-4 and its ligands, VEGF-C and VEGF-D, in tumor
lymphangiogenesis and lymphatic metastasis in multi-
ple solid tumor types.42
VEGF-A promotes proliferation of endothelial cells
and monocytes. It also induces endothelial cell expres-
sion of antiapoptotic protein bcl-2 and thus acts as a
43
survival factor for endothelial-like cells. In addition,
44
VEGF can inhibit maturation of dendritic cells. Thus,
VEGF-A enriches the pool of cells that can contribute
to angiogenesis but also depletes the reservoir of cells
involved in host immune response, potentially lead-
ing to immunosuppression. In addition to these roles,
VEGF-A is a vascular permeability factor, producing
leakage of plasma proteins that result in the formation
of extravascular fibrin gel, which provides a substrate
for endothelial and tumor cell growth.45
Transcriptional Regulation of VEGF
Nuclear factor-κB (NF-κB) and activated protein (AP-
1) have been implicated in the control of VEGF tran-
scription.
46,47
Inhibition of NF-κB activity reduced
VEGF expression as well as tumorigenicity and angio-
48,49
genesis in several cancer models. AP-1 has been
shown to act synergistically with NF-κB to promote IL-
50
8 transcription and to play a role in IL-8 expression in
51
response to hypoxia. AP-1 has also been shown to
play a positive role in the induction of VEGF expres-
sion by hypoxia, transforming growth factor–β, and ac-
idosis.52 Several factors that can potentiate VEGF pro-
duction include fibroblast growth factor 4, platelet-
derived growth factor, tumor necrosis factor–α, trans-
forming growth factor–β, keratinocyte growth factor,
insulin-like growth factor, interleukin-1β, and IL-6.53
Moreover, all these cytokines are regulated by the tran-
scription factors NF-κB and AP-1 directly or indirectly.
VEGF-Mediated Antiangiogenic Therapy
Direct evidence of the role of VEGF in tumorigenesis
was provided by studies with monoclonal antibodies
against VEGF. Inhibition of growth of a wide variety of
human tumor cell lines (carcinomas, sarcomas, and
gliomas) in mice by antihuman VEGF monoclonal an-
tibodies correlated with nearly complete suppression
of tumor-associated angiogenesis.54 Several inhibitors
of angiogenesis are currently in clinical development
for the treatment of cancer. The agents include anti-
bodies such as neutralizing anti-VEGF antibody, an an-
55
tibody against VEGF receptors. Many of these agents
block angiogenesis either by blocking the VEGF re-
ceptor or by neutralizing VEGF itself. There are indi-
rect angiogenic inhibitors such as HER-2 neutraliz-
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VEGF and Receptors
ing antibody (trastuzumab), which indirectly inhibits
angiogenesis because its target (HER-2) stimulates
55
VEGF expression.
The in vivo regulation of a VEGF gene by RNA
interference has been blocked by the ability of tumor
cells to secrete high levels of VEGF, which could
diminish or prevent the triggering of therapeutic
resistance of tumor cells. It has been reported that
small interfering RNA–mediated inhibition of VEGF
severely limited the tumor resistance to antiangio-
genic thrombospondin-1 and slowed tumor vasculari-
zation and growth.56 Antiangiogenesis therapies di-
rected against the VEGF-VEGF-R kinase axes through
a variety of approaches have been a promising and
well-validated therapeutic approach and are under
active evaluation for their safety and efficacy in multi-
ple clinical trials. In addition, several orally active
small molecule inhibitors of specific VEGFR-kinases,
including PTK-787 (vatalinib), PKC-412 (midstaurin),
ZD-6474, SU-11248, and CP-547/632, are currently
under clinical evaluation for the development of anti-
angiogenic molecules.38 The pharmacological, phar-
macokinetic, and tolerability profile of these agents in
preclinical studies is compatible with chronic admin-
istration against a variety of solid tumors.
Effect of Natural Products on
VEGF-Mediated Angiogenesis
A variety of studies have suggested the inhibitory role
of natural product and dietary compounds on tumor-
specific angiogenesis. Our laboratory has reported
the effect of Tinospora cordifolia (Willd.) Miers (Meni-
spermaceae) on the inhibition of VEGF production
and tumor-specific angiogenesis in mice.57 Several
other studies of the impact of natural products on
tumor-specific angiogenesis are being conducted in
our laboratory. Resveratrol, a polyphenolic com-
pound found in grapes and other fruits, has been re-
ported to inhibit VEGF-induced angiogenic effects in
human umbilical vein endothelial cells.58 The inhibi-
tory effect of indole-3-carbinol, a major indole metab-
olite in cruciferous vegetables, on cell proliferation
and in vitro markers of angiogenesis in phorbol my-
ristate acetate–stimulated endothelial cells has been
reported.59
Capsaicin, a natural product present in Capsicum
species, inhibits VEGF expression and related angio-
60
genesis in in vitro as well as in vivo models. It was
found that sanguinarine, a benzophenanthridine
alkaloid derived from the root of Sanguinaria cana-
densis L. (Papaveraceae), strongly suppressed basal
and VEGF-induced Akt phosphorylation, which inhib-
its angiogenesis.61 Ellagic acid, another natural poly-
phenolic compound found in fruits and nuts, inhibits
319
 Pradeep et al
VEGF-induced phosphorylation of VEGFR-2 and
62
angiogenesis. The antiangiogenic potency of vitamin
E, catechin, and green tea catechins has also been
63,64
reported previously. These agents appear to target
common mechanisms of tumor angiogenesis that may
permit identification of critical targets for antiangio-
genic therapy and antiangiogenic chemoprevention.
Thus, compounds that inhibit the effects of VEGF
have the potential to provide a novel, effective, and
well-tolerated therapy for solid tumors. These agents
may also provide a new therapeutic approach for the
treatment of other diseases in which angiogenesis
plays an important role.
Conclusion
This review discusses the multiple roles of VEGF in
tumor-specific angiogenesis. VEGF, in its function as a
master regulator of the angiogenic switch, promotes
angiogenesis through multiple and complementary
mechanisms. Not only does it promote the recruit-
ment and proliferation of endothelial cells and their
precursors within the tumor, but it also furnishes
nearby host vessels for the necessary plasticity to sup-
port angiogenesis in developing tumors.
Acknowledgment
The authors apologize to those investigators whose ex-
perimental work has only been cited indirectly in this
article because of space limitations.
References
1. Folkman J. Angiogenesis in cancer, vascular, rheumatoid and
other diseases. Nat Med. 1995;1:27-31.
2. Kerbel RS. Tumor angiogenesis: past, present and the near
future. Carcinogenesis. 2000;21:505-515.
3. Griffin RJ, Williams BW, Wild R, Cherrington JM, Park H, Song
CW. Simultaneous inhibition of the receptor kinase activity of
vascular endothelial fibroblast and platelet derived growth fac-
tors suppresses tumor growth and enhances tumor growth and
enhances tumor radiation response. Cancer Res. 2002;62:1702-
1706.
4. Stacker SA, Achen MG, Jussila L, Baldwin ME, Alitalo K.
Lymphangiogenesis and cancer metastasis. Nat Rev Cancer.
2002;2:573-583.
5. Ramanujam S, Koenig GC, Pdera TP, Stoll BR, Jain RK. Local
imbalance of proangiogenic and antiangiogenic factors: a
potential mechanism of local necrosis and dormancy in tumors.
Cancer Res. 2000;60:1442-1448.
6. Lu C, Tanigawa N. Spontaneous apoptosis is inversely related to
intratumoral microvessel density in gastric carcinoma. Cancer
Res. 1997;57:221-224.
7. Benjamin LE, Golijanin D, Itin A. Selective ablation of imma-
ture blood vessels in established human tumors follows vascular
endothelial growth factor withdrawal. J Clin Invest. 1999;103:
159-165.
8. Cristofanilli M, Charnasangavej C, Hortobagyi GN. Angiogene-
sis modulation in cancer research: novel clinical approaches.
Nat Rev Drug Discov. 2002;1:415-426.
320
Downloaded from ict.sagepub.com at RYERSON UNIV on April 26, 2015
9. Reinmuth N, Liu W, Jung YD, et al. Induction of VEGF in
perivascular cell defines a potential paracrine mechanism for
endothelial cell survival. FASEB J. 2001;15:1239-1241.
10. Ferrara N. Vascular endothelial growth factor and the regula-
tion of angiogenesis. Recent Prog Horm Res. 2000;55:15-35.
11. Clauss M. Molecular biology of the VEGF and the VEGF recep-
tor family. Semin Thromb Hemostat. 2000;26:561-569.
12. Veikkola T, Karkkainen M, Clausson L. Regulation of angio-
genesis via vascular endothelial growth factor receptors. Cancer
Res. 2000;60:203-212.
13. Yancopoulos GD. Vascular-specific growth factors and blood
vessel formation. Nature. 2000;407:242-248.
14. Shima DT, Mailhos C. Vascular developmental biology: getting
nervous. Curr Opin Gen Dev. 2000;10:536-542.
15. Robinson CJ, Stringer SE. The splice variants of vascular endo-
thelial growth factor (VEGF) and their receptors. J Cell Sci. 2001;
114:853-865.
16. Hiratsuka S, Maru Y, Okada A, Seiki M, Noda T, Shibuya M. In-
volvement of Flt-1 tyrosine kinase (vascular endothelial growth
factor receptor-1) in pathological angiogenesis. Cancer Res.
2001;61:1207-1213.
17. Ferrara N, Park JE. Vascular endothelial growth factor (VEGF).
In: Nicola N, ed. Guidebook to Cytokines and Their Receptors. New
York, NY: Oxford University Press; 1994:232-234.
18. Rahim N, Dayanir V, Lashkari K. Receptor chimeras indicate
that the vascular endothelial growth factor receptor-1 modu-
lates mitogenic activity of VEGFR-2 in endothelial cells. J Biol
Chem. 2000;275:16986-16992.
19. Gille H, Kowalski J, Li B, et al. Analysis of biological effects and
signaling properties of Flt-1 (VEGFR-1) and KDR (VEGFR-2). J
Biol Chem. 2001;276:3222-3230.
20. Achen MG. Vascular endothelial growth factor D (VEGF-D) is
a ligand for the tyrosine kinases VEGF receptor 2 (Flk1) and
VEGF receptor 3 (Flt4). Proc Natl Acad Sci U S A. 1998;95:548-
553.
21. Carmeliet P, Collen D. Role of vascular endothelial growth fac-
tor and vascular endothelial growth factor receptors in vascular
development. Curr Top Microbiol Immunol. 1999;237:133-158.
22. Neufeld G. Vascular endothelial growth factor (VEGF) and its
receptors. FASEB J. 1999;13:9-22.
23. White JD, Hewette PW, Kosuge D, et al. Vascular endothelial
growth factor-D expression is an independent prognostic
marker for survival in colorectal carcinoma. Cancer Res. 2002;
62:1669-1675.
24. Ferrara N. Role of vascular endothelial growth factor in regula-
tion of angiogenesis. In: Teicher BA, ed. Antiangiogenesis in Can-
cer Therapy. Totowa, NJ: Humana Press; 1999:119-142.
25. Maxwell PH, Pugh CW, Ratcliffe PJ. Activation of the HIF path-
way in cancer. Curr Opin Genet Dev. 2001;11:293-299.
26. Harris AL. Hypoxia: a key regulatory factor in tumor growth.
Nat Rev Cancer. 2002;2:38-47.
27. Levy AP, Levy NS, Wegner S, Goldberg MA. Transcriptional reg-
ulation of the rat vascular endothelial growth factor gene by
hypoxia. J Biol Chem. 1995;270:13333-13340.
28. Jiang BH, Agani F, Passaniti A, Semenza GL. v-src induces
expression of hypoxia-inducible factor 1 (HIF-1) and transcrip-
tion of genes encoding vascular endothelial growth factor and
enolase 1: involvement of HIF-1 in tumor progression. Cancer
Res. 1997;57:5328-5335.
29. Levy NS, Chung S, Furneaux H, Levy AP. Hypoxic stabiliza-
tion of vascular endothelial growth factor mRNA by the RNA-
binding protein HuR. J Biol Chem. 1998;273:6417-6423.
30. Grunstein J, Masbad JJ, Hickey R, Giordano F, Johnson RS.
Isoforms of vascular endothelial growth factor act in a coordi-
nate fashion to recruit and expand tumor vasculature. Mol Cell
Biol. 2000;20:7282-7291.
INTEGRATIVE CANCER THERAPIES 4(4); 2005

31. Viscotini RP, Richardson CD, Sato TN. Orchestration of angio-
genesis and arteriovenous contribution by angiopoetins and
vascular endothelial growth factor (VEGF). Proc Natl Acad Sci U S
A. 2002;99:8219-8224.
32. Benjamin LE, Hemo I, Keshet E. A plasticity window for blood
vessel remodeling is defined by pericyte coverage of the per-
formed endothelial network and is regulated by PDGF-B and
VEGF. Development. 1998;125:1591-1598.
33. Stone J. Roles of vascular endothelial growth factor and
astrocyte degeneration in the genesis of retinopathy of pre-
maturity. Invest Ophthal Vis Sci. 1996;37:290-299.
34. Carmeleit P, Jain RK. Angiogenesis in cancer and other diseases.
Nature. 2000;407:249-257.
35. Korpelainen EI, Alitalo K. Signaling angiogenesis and
lymphangiogenesis. Curr Opin Cell Biol. 1998;10:159-164.
36. Griffioen AW, Molema G. Angiogenesis: potentials for pharma-
cologic intervention in the treatment of cancer, cardiovascular
diseases and chronic inflammation. Pharmacol Rev. 2000;52:237-
268.
37. Neufield G, Cohen T, Gengronivitch S, Polotruck Z. Vascular
endothelial growth factor (VEGF) and its receptors. FASEB J.
1999;13:9-22.
38. Ruggeri B, Singh J, Gingrich D, et al. CEP 7055: a novel orally
active pan inhibitor of vascular endothelial growth factor recep-
tor tyrosine kinases with potent antiangiogenic activity and
antitumor efficacy in preclinical model. Cancer Res. 2003;63:
5978-5991.
39. Lee CG, Heijin M, Tomaso E, et al. Anti-vascular endothelial
growth factor treatment augments tumor radiation response
under normoxic or hypoxic conditions. Cancer Res. 2000;60:
5565-5570.
40. Wang H, Keiser JA. Vascular endothelial growth factor up-
regulates the expression of matrix metalloproteinases in vascu-
lar smooth muscle cells: role of Flt-1. Circ Res. 1998;83:832-840.
41. Hattori K, Heissig B, Wu Y, et al. Placental growth factor recon-
stitutes hematopoiesis by screening VEGFR(+) stem cells from
bone marrow microenvironment. Nat Med. 2002;8:841-849.
42. Beasley NJP, Prevo R, Banerji S, et al. Intratumoral lymphangio-
genesis and lymph node metastasis in head and neck cancer.
Cancer Res. 2002;62:1315-1320.
43. Ferrara N. VEGF: an update on biological and therapeutic
aspects. Curr Opin Biotechnol. 2000;11:617-624.
44. Fernandez PB, Lucibello FC, Zuzarte M, et al. Dendritic cells
derived from peripheral monocytes express endothelial mark-
ers and in the presence of angiogenic growth factors differenti-
ate into endothelial-like cells. Eur J Cell Biol. 2001;80:99-110.
45. Dvorak AM, Feng D. The vesiculo-vacoular organelle: a new
endothelial cell permeability organelle. J Histochem Cytochem.
2001;49:99-110.
46. Bancroft CC, Chen Z, Dong G, et al. Coexpression of pro-
angiogenic factors IL-8 and VEGF by human head and neck
squamous cell carcinoma involves coactivation by MEK-MAPK
and IKK-NF-κB signal pathways. Clin Cancer Res. 2001;7:435-442.
47. Chua CC, Hamdy RC, Chua BH. Mechanism of transform-
ing growth factor-β induced expression of vascular endothelial
growth factor in murine osteoblastic MC3T3-E1 cells. Biochim
Biophys Acta. 2000;1497:69-76.
48. Huang S, Pettaway CA, Uehra H, Bucana CD, Fidler IJ. Block-
ade of NF-kappaB activity in human prostate cancer cells is
INTEGRATIVE CANCER THERAPIES 4(4); 2005
Downloaded from ict.sagepub.com at RYERSON UNIV on April 26, 2015
VEGF and Receptors
associated with suppression of angiogenesis, invasion, and
metastasis. Oncogene. 12;20:4188-4197.
49. Huang S, Robinson JB, Deguzman A, Bucana CD, Fidler IJ.
Blockade of nuclear factor-kappaB signaling inhibits angio-
genesis and tumorigenicity of human ovarian cancer cells by
suppressing expression of vascular endothelial growth factor
and interleukin-8. Cancer Res. 2000;1:5334-5339.
50. Mastronarde JG, Monick MM, Mukaida N, Matushima K,
Hunninghake GW. Activator protein-1 is the preferred tran-
scription factor for cooperative interaction with nuclear factor-
kappaB in respiratory syncytial virus-induced interleukin-8
gene expression in airway epithelium. J Infect Dis. 1998;7:1275-
1281.
51. Shi Q, Le X, Abbruzzese JL, et al. Cooperation between tran-
scription factor AP-1 and NF-κB in the induction of interleukin-
8 in human pancreatic adenocarcinoma cells by hypoxia. J Inter-
feron Cytokine Res. 1999;19:1363-1371.
52. Damert A, Ikeda E, Riasu W. Activator-protein-1 binding poten-
tiates the hypoxia inducible factor-1-mediated hypoxia-induced
transcriptional activation of vascular endothelial growth factor
expression in C6 glioma cells. Biochem J. 1997;327:419-423.
53. Neufeld G, Cohen T, Gengrinovitch S, Poltrok Z. Vascular endo-
thelial growth factor (VEGF) and its receptors. FASEB J. 1999;
13:9-22.
54. Gerber HP, Kowalski J, Sherman D. Complete inhibition of
rhabdosarcoma xenograft growth and neovascularization re-
quires blockade of both tumor and host vascular endothelial
growth factor. Cancer Res. 2000;60:6253-6258.
55. Folkman J. Role of angiogenesis in tumor growth and metasta-
sis. Semin Oncol. 1998;29:15-18.
56. Filler S, Courtin A, Ait-Si-Ali S, Guglielmi J, Merle C, Hard-
Bellan A. SiRNA mediated inhibition of vascular endothelial
growth factor severely limits tumor resistance to antiangiogenic
thrombospondin-1 and slows tumor vascularization and
growth. Cancer Res. 2003;63:3919-3922.
57. Leyon PV, Kuttan G. Effect of Tinospora cordifolia on cytokine
profile of angiogenesis-induced mice. Int Immunopharmacol.
2004;4:1569-1575.
58. Lin MT, Yen M-L, Chen Y-H. Inhibition of VEGF-induced angio-
genesis by resveratrol through interruption of Src-dependent
vascular endothelial cadherin tyrosine phosphorylation. Mol
Pharmacol. 2003;64:1029-1036.
59. Wu H-T, Lin SH, Chen Y-H. Inhibition of cell proliferation and
in vitro markers of angiogenesis by indole metabolite in
cruciferous vegetables. J Agric Food Chem. 2005;53:5164-5169.
60. Min JK, Han KY, Kim EC, et al. Capsaicin inhibits in vitro and in
vivo angiogenesis. Cancer Res. 2004;64:644- 651.
61. Eun JP, Koh GY. Suppression of angiogenesis by the plant alka-
loid sanguinarine. Biochem Biophys Res Commun. 2004;317:618-
624.
62. Labrecque L, Larry S, Chapus A, et al Combined inhibition of
PDGF and VEGF receptors by ellagic acid, a dietary-derived phe-
nolic compound. Carcinogenesis. 2005;64:821-826.
63. Miyazawa T, Tsuzuki T, Nakagawa K, Igarashi M. Antiangiogenic
potency of vitamin-E. Ann N Y Acad Sci. 2004;1031:401-404.
64. Kondo T, Ohta T, Igura K, Hara Y, Kaji K. Tea catechins inhibit
angiogenesis in vitro, measured by human endothelial cell
growth, migration and tube formation through inhibition of
VEGF receptor binding. Cancer Lett. 2002;180:139-144.
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