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Tumor Biology |
Division of Biochemistry, Department of Oncology, Biomedical Research Center, Osaka University Medical School, Suita, Osaka 565-0871, Japan [K. K., K. M., T. N.]; Department of Surgery 1, Kyushu University Faculty of Medicine, Maidashi, Fukuoka 812-8582, Japan [K. K., K. D., M. T.]; and Department of Surgery 1, Fukuoka University School of Medicine, Nanakuma, Fukuoka 814-0180, Japan [H. S.]
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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HGF is composed of the 
chain, which contains a
NH2-terminal hairpin and four kringle
domains and the catalytically inactive serine protease-like ß-chain
(2)
. Recently we prepared an antagonist for HGF by
proteolytic digestion of HGF (17)
, and this
HGF-antagonist, called HGF/NK4 (or NK4), is composed of the
NH2-terminal hairpin domain and subsequent four
kringle domains of the -subunit of HGF. NK4 binds to the c-Met/HGF
receptor, but does not induce tyrosine phosphorylation of c-Met
(17)
. NK4 competitively inhibits biological events driven
by HGF-Met receptor coupling, including the invasion of distinct types
of tumor cells (17
, 18)
. On the other hand, NK1 and NK2,
previously characterized HGF variants, have partial agonist and
antagonist activity and elicit motility and the invasion of tumor cells
and endothelial cells (19, 20, 21, 22)
.
Angiogenesis, the formation of new blood vessels from preexisting blood vessels, is a critical process involved in embryonic development, tissue regeneration, and pathological conditions such as tumorigenesis and diabetic retinopathy (23 , 24) . Many investigators reported the essential role of angiogenesis during tumor progression (23 , 25) . Studies led to the thesis that angiogenesis is regulated by a balance between angiogenic and angioinhibitory factors (25 , 26) . In the activated endothelium, angiogenic growth factors predominate, whereas vascular quiescence is achieved by the dominance of angioinhibitory factors including angiostatin, endostatin, thrombospondin, platelet factor IV, the NH2-terminal fragment of prolactin, etc. (25) . Physiological and pathophysiological roles of angioinhibitory factors, as well as mechanisms by which these polypeptides inhibit angiogenesis, are largely unknown; however, several angiogenesis inhibitors have been shown to inhibit tumor growth and metastasis (27, 28, 29, 30) , and there are angiogenesis inhibitors under clinical trials for cancer treatment (29, 30, 31, 32) .
We now have evidence that NK4 is an angiogenesis inhibitor. NK4 not only antagonizes HGF-induced angiogenesis but also abrogates the angiogenesis induced by other angiogenic inducers. The antiangiogenic activity of NK4 is likely to be exhibited through a mechanism distinct from its initially characterized potential to act as an HGF-antagonist: the blockade of HGF-c-Met coupling. We report here that NK4 suppresses tumor angiogenesis, growth, and metastasis in mice.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Cell Culture.
Human adult dermal microvascular endothelial cells and human
lung-derived microvascular endothelial cells were purchased from
Clonetics (San Diego, CA) and grown in EBM-2 supplemented with 5% FBS
and endothelial cell growth supplements (Clonetics). Human pulmonary
artery endothelial cells, purchased from KURABO (Osaka, Japan),
were cultured in endothelial growth medium (EGM; KURABO). Rat coronary
endothelial cells were isolated as described (35)
. Culture
plates for endothelial cells were coated with 0.1% gelatin (Difco,
Detroit, MI). Human dermal fibroblasts initially proliferated outward
from dermal tissue obtained during plastic surgery. These cells were
used within passage number 4–8. MDCK epithelial cells
were a generous gift from Dr. R. Montesano (University of Geneva,
Geneva, Switzerland). NIH3T3 fibroblasts, LLC cells, and Jyg-MC
cells were obtained from RIKEN Cell Bank (Tsukuba, Japan). Rat
coronary endothelial cells, human fibroblasts, MDCK, and NIH3T3 cells
were cultured in DMEM supplemented with streptomycin, penicillin, and
10% FBS.
Cell Proliferation, Migration, and Invasion Assay.
Endothelial cells were plated at 5 x 103
cells/cm2 onto
gelatinized 24-well tissue culture plates and cultured for 24 h.
The medium was replaced with 0.5 ml of EBM-2 containing 5% FBS, and
cells were cultured in the absence or presence of NK4, 10 ng/ml HGF, 3
ng/ml bFGF, 10 ng/ml VEGF, or their combinations. After 72 h,
cells were dispersed by trypsin and counted by Coulter counter. To
measure the proliferation of nonendothelial cells, human dermal
fibroblasts, LLC cells, and Jyg-MC cells were plated at 5 x 103
cells/cm2,
whereas NIH3T3 and MDCK cells were plated at 2.5 x 103
cells/cm2, on 24-well
plates and cultured for 24 h. The medium was replaced with DMEM
supplemented with 5% FBS and 10 ng/ml bFGF, test samples were added,
and the cells were then cultured for 72 h.
Migration of endothelial cells was evaluated using a modified Boyden chamber assay, as described (12 , 36) . The cells were serum-starved in EBM-2 medium for 12 h and plated at 12 x 104 cells/cm2 onto the polycarbonate filter with 5-µm pores (Costar, Cambridge, MA) coated with 13.4 µg/ml fibronectin (Orgagnon Teknika Corp., West Chester, PA). Test samples were added to the medium in the outer cup, and the cells were cultured for 5 h. The number of the cells which migrated to the undersurface of the filter was quantified by counting cells in five randomly selected microscopic fields (x200) in each well. In-vitro invasion of carcinoma cells was measured using a Matrigel invasion chamber (Becton Dickinson, Bedford, MA), as described (12) .
Immunoprecipitation and Western Blot.
Tyrosine phosphorylation of c-Met or KDR was analyzed as
described elsewhere (18)
. Briefly, human adult dermal
microvascular endothelial cells were grown on 100-mm plates and
serum-starved overnight before treatment for 10 min with various
concentrations of NK4 with or without bFGF, VEGF, or HGF (10 ng/ml
each). Cell lysates were prepared, and equivalent amounts of protein
were incubated overnight with a monoclonal antibody against
phosphotyrosine (PY99; Santa Cruz Biotechnology, Santa Cruz, CA) and
then incubated for 2 h with protein G-Sepharose. The
immunoprecipitates were separated by 6% SDS-PAGE and electroblotted
onto polyvinylidene difluoride membranes, and the proteins were
probed with anti-c-Met antibodies (C-12; Santa Cruz
Biotechnology) or anti-KDR/Flk-1 antibodies (C-1158; Santa Cruz
Biotechnology). Immunoreactive bands were visualized by the enhanced
chemiluminescence system (Amersham). For detection of phosphorylated
ERK1/2 (p44/42 MAPK), total cell lysates were separated
by 12% SDS-PAGE and transferred to polyvinylidene difluoride
membranes. Blots were developed with phosho-specific antibodies
against ERK1/2 (New England Biolabs, Beverly, MA). After being
"stripped," the membranes were reprobed with antibody against
ERK1/2 (Santa Cruz Biotechnology) as a loading control.
Tumor Study in Mice.
Male nude mice 6- to 8-weeks-old (BALB/c nu/nu; Japan SLC, Inc.,
Hamamatsu, Japan) were s.c. implanted with 1 x 106 LLC cells or 5 x 106 Jyg-MC cells. After 4 days, an osmotic pump
(Alzet 2002; Alza Corp., Palo Alto, CA) containing NK4 or BSA or saline
alone was surgically implanted near the tumor, and NK4 or BSA solution
or saline alone was continuously infused for 10 or 14 days into the
s.c. region near the tumor mass. The size of tumors in all groups was
measured using a dial caliper, and the volume of tumors was determined
using the formula width2 x length x 0.52. To analyze external lung metastases, mice
were infused with NK4 solution or saline alone for 14 days, as
described above, then the mice were killed on day 28 after tumor
implantation.
Tumor tissues were fixed in Carnoy’s fixative for 4 h or overnight in 70% ethanol at 4°C and embedded in paraffin according to standard histological procedures. For blood vessel staining, tissue sections fixed in Carnoy’s fixative were pretreated with 5 µg/ml Proteinase K at 37°C for 15 min and incubated with antibody against von Willebrand factor (Dako, Glostrup, Denmark). The sections were sequentially incubated with biotin-labeled secondary antibodies and avidin-biotin peroxidase complex, as described (37) . The number of blood vessels was counted under a light microscope at x200 magnification using at least 20 randomly selected fields per section. For detection of proliferating cells or apoptotic cells, tissue sections fixed in 70% ethanol were analyzed as described elsewhere (18) . Statistical analyses were performed with unpaired Student’s t test (two-tailed). P < 0.05 was considered to be statistically significant.
In Vivo Angiogenesis Assay.
Antiangiogenic activity of NK4 on CAM was assayed, as described
(29
, 38) . Briefly, fertilized white Leghorn chicken eggs
were incubated at 37°C for 5 days, and a methylcellulose disk
containing test material was placed within a sterilized silicon ring on
CAM (38)
. The eggs were incubated at 37°C for 2 days.
The white fat emulsion (Intralipos; WelFide Co., Osaka, Japan) was
injected into the chorioallantois, and the vascular networks in the
CAMs were blindly scored by two independent investigators for the
presence or absence of an avascular zone (>3 mm in diameter)
surrounding the implant. Statistical analyses were performed
with the Fisher’s exact probability test. P < 0.05 were
considered to be statistically significant.
The antiangiogenic activity of NK4 was also assayed in the rabbit
cornea, as described (39, 40, 41)
. Slow-releasing pellets were
prepared by incorporating HGF, bFGF and/or NK4 into a 20-µl casting
solution of an ethylene-vinyl acetate copolymer (EV40) in 5% methylene
chloride. A microsurgical pocket (2 x 4 mm) was
produced in the lower half of each eye of male albino Japanese rabbits
such that a peripheral pocket ended at 2 mm from the limbus. A single
pellet was deposited in the bottom of the pocket. Eyes were examined in
a blind manner by slit-lamp microscopy every other day. An angiogenic
response was scored positive when blood vessels from the limbal plexus
occurred and capillaries progressed to reach the implanted pellet
within 6–8 days, as described (40)
. Statistical analysis
was performed with the 
2 test. P
< 0.05 was considered to be statistically significant.
Data Analysis.
Statistical analyses were performed with unpaired Student’s
t test (two-tailed) unless mentioned otherwise in the text.
Differences were considered to be statistically significant at
P < 0.05.
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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, and not shown). On the other hand, both HGF and bFGF stimulated
invasion of tumor cells, whereas NK4 specifically antagonized the
invasion of these tumor cells induced by HGF but not by bFGF (Fig. 1B)
. When LLC and Jyg-MC cells are inoculated s.c. into
athymic mice, both tumors formed nodules (20–40
mm3
in volume) 4 days after implantation, and
metastatic nodules in lung surface become visible 3–4 weeks after
implantation.
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). Lung metastasis was strongly suppressed by
NK4-treatment in a dose-dependent manner; the number of lung metastases
in control mice (12.9 ± 1.35) decreased to 54.8%
(P < 0.05) and 9.46%
(P < 0.0001) in NK4-treated mice at 10
µg/day and 25 µg/day, respectively (Fig. 2B)
. Similarly,
NK4 (25 µg/day) also suppressed primary tumor growth and lung
metastasis of Jyg-MC to 53.6% (P < 0.05)
and 35.3% (P < 0.05), respectively
(n = 4/group; Fig. 2
). Infusion of BSA in
saline at 25 µg/day for 2 weeks did not significantly change the
tumor volume and number of lung metastases of both tumors (not shown).
Although NK4 inhibited tumor metastasis, the result raised the question
as to how NK4 suppresses growth of these primary tumors, inasmuch as
HGF and NK4 have no direct effect on the proliferation of tumor cells.
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. Thus, we speculated that NK4 suppresses tumor growth by inhibiting
the host compartment, such as angiogenesis, rather than the tumor-cell
compartment. To examine this possibility, we measured the microvessel
density in tumor tissues by von Willebrand factor
immunostaining. The number of von Willebrand
factor-positive vessels in NK4-treated tumors decreased to 55.8% of
that of control tumors (P < 0.001), and some
tumor vessels in NK4-treated mice were disrupted and had a short
diameter (Fig. 3)
. Furthermore, TUNEL assay showed that
NK4-treatment led to a 2.25-fold increase in the number of apoptotic
cells (P < 0.001; Fig. 3
). These
findings are consistent with previous studies which showed that
an angiogenesis inhibitor suppresses tumor growth by increasing
apoptosis of tumor cells (37
, 42)
. Thus NK4 seems to
suppress primary tumor growth mainly through inhibition of tumor
angiogenesis.
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To determine whether NK4 has similar inhibitory effects on other
endothelial and nonendothelial cells, we examined the effects of NK4 on
the proliferation of several types of cells (Fig. 4B)
. Three distinct types of endothelial cells (human
lung-derived microvascular endothelial cells, human pulmonary artery
endothelial cells, and rat coronary endothelial cells) were cultured in
the presence of bFGF with or without NK4. Dose-dependent growth
inhibition by NK4 was seen in these three types of endothelial cells,
whereas effectiveness differed somewhat for each cell type; the most
potent inhibitory effect was seen with human microvascular endothelial
cells. A similar inhibitory effect of NK4 was seen when these
endothelial cells were cultured in the presence of VEGF (not shown). On
the other hand, the proliferation of nonendothelial cells (human dermal
fibroblasts, NIH3T3 mouse fibroblasts, MDCK canine renal epithelial
cells), which were stimulated by bFGF, was not suppressed by NK4 at all
(Fig. 4B)
. Likewise, NK4 had no inhibitory effect on
proliferation of these nonendothelial cells stimulated by 10% FBS (not
shown).
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. Endothelial cells
were seeded on a filter membrane coated with fibronectin, and the
number of cells that migrated through the membrane was measured.
Migration of endothelial cells was stimulated by bFGF, VEGF, and HGF,
whereas migration of the endothelial cells driven by these growth
factors was strongly inhibited by NK4, mostly to the basal level at 300
nM NK4. Taken together, NK4 has an inhibitory
effect on endothelial cell growth and migration stimulated by bFGF and
VEGF as well as by HGF, and the antiproliferative activity is likely to
be specific to endothelial cells.
Effects of Neutralizing HGF-antibody on Endothelial Growth and
Migration.
Although NK4 inhibited endothelial growth and migration
stimulated by bFGF and VEGF, its antagonizing activity for HGF might be
involved in the endothelial-inhibitory action of NK4. To address this
issue, we tested the effect of neutralizing antibody on the growth and
migration of endothelial cells (Fig. 4D)
. In contrast to
NK4, anti-HGF antibody did not inhibit bFGF- and VEGF-induced
endothelial growth. On the other hand, the stimulatory effect of HGF on
endothelial cell growth was almost completely inhibited by anti-HGF
antibody, equally to NK4. Similarly, endothelial cell migration
mediated by bFGF and VEGF was not affected by anti-HGF antibody (not
shown).
Effects of NK4 on Receptor Tyrosine Phosphorylation and ERK1/2
Activation.
To investigate the possibility that the binding of NK4 to the Met
receptor may modify ligand-receptor interaction and subsequent
activation of signaling events, or that NK4 itself may interfere with
ligand-dependent activation of the receptor for angiogenic growth
factors, we examined the tyrosine phosphorylation state of c-Met in
endothelial cells. HGF, but not bFGF or VEGF, specifically
activated c-Met in endothelial cells by increasing the phosphorylation
on tyrosine residues, as shown by immunoprecipitation and Western
blotting (Fig. 5A)
. NK4 inhibited HGF-induced phosphorylation of the Met
receptor. On the other hand, NK4 alone, in concentrations up to 1000
nM (67 µg/ml), did not induce tyrosine
phosphorylation of c-Met, and NK4 (300 nM) also
failed to activate c-Met in the presence of bFGF or VEGF.
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. In contrast with the
blockade of HGF-Met coupling by NK4, the VEGF-induced phosphorylation
of KDR was not inhibited by NK4. Because activation of ERK1/2 (p44/42
MAPK) is closely involved in the process of angiogenesis induced by
various angiogenic stimuli, we next analyzed activation of ERK1/2 (Fig. 5C)
. HGF, bFGF, and VEGF induced prominent activation of
ERK1/2, whereas NK4 did not inhibit the bFGF- or VEGF-induced
activation of ERK1/2 but did inhibit HGF-induced ERK1/2 activation.
Therefore, NK4 does not inhibit early intracellular signaling events
driven by bFGF and VEGF, yet NK4 inhibits bFGF- and VEGF-induced
angiogenic responses in endothelial cells.
In-Vivo Antiangiogenic Activity of NK4.
To study the angiostatic activity of NK4 in vivo, NK4 was
tested using CAM (Fig. 6A)
. In controls, CAMs with avascular zones were never
found. However, NK4 inhibited new blood vessel formation in a
dose-dependent manner as determined by the formation of avascular zones
(Fig. 6A)
; at a dose of 60 µg NK4/disk, avascular zones
were seen in 61.5% of embryos (P < 0.01).
Importantly, the inhibitory activity of NK4 on angiogenesis in chick
CAMs diminished when NK4 was tested after heat-treatment. BSA (60
µg/disk) had no inhibitory effect on angiogenesis in the CAM assay.
The results indicate that NK4 has antiangiogenic activity in
vivo as well as in vitro.
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. Application of
100 ng of bFGF induced neovascularization in 70% of the
corneas (7 of 10), whereas simultaneous application of NK4
dose-dependently inhibited bFGF-induced neovascularization. NK4 at 1000
ng significantly inhibited bFGF-induced neovascularization to 22%
(P < 0.05). Heat-treated NK4 had no effect
on bFGF-induced neovascularization, and NK4 alone had no effect on
neovascularization in the cornea. Application of HGF in the cornea
assay induced neovascularization and NK4 had inhibitory effects on
HGF-induced neovascularization (not shown). |
DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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With regard to the structure and function of NK4, it is noteworthy that NK4 has a significant structural similarity with angiostatin, a potent angiogenesis inhibitor (29) . Angiostatin is an internal fragment of plasminogen that encompasses the first four kringle domains, and the amino acids sequence homology between four kringles of NK4 and angiostatin reaches 47%. Previous studies showed that individual kringle domains of angiostatin, the fifth kringle domain of plasminogen, and the prothrombin kringle-2 domain have antiangiogenic activity (44, 45, 46) . Our preliminary results indicated that antiangiogenic activity of NK4 seems to reside within kringle domains.4 These results suggest the possibility that a structural motif conserved in some kringle domains may be involved in inhibiting angiogenesis. However, it is equally probable that NK4 inhibits angiogenesis through a mechanism distinct from angiostatin, because NK4 inhibits DNA synthesis and induces cell cycle arrest,5 whereas angiostatin increases endothelial apoptosis without inhibiting DNA synthesis (34 , 47) .
Of particular importance in the present study is the in vivo
inhibition of tumor metastasis by NK4. The metastatic cascade
is composed of multiple steps: (a) induction of
angiogenesis; (b) dissociation of tumor cells;
(c) invasion through the extracellular matrix;
(d) intravasation; (e) transport in the
circulation; (f) arrest in a distant capillary bed, and
(g) extravasation followed by the establishment of secondary
foci (48)
. In our experimental model, inhibition of lung
metastasis might be the result of suppression of early events
(i.e., angiogenesis and invasion) in the primary tumors,
because NK4-treatment was terminated 10 days before evaluation of lung
metastasis, and this no-treatment period might have allowed for the
growth of secondary tumors (Fig. 2A)
. Considering the
bifunctionality of NK4 (HGF antagonist and angiogenesis inhibitor), we
can raise the possibility that NK4 may have suppressed lung metastasis
by both actions: one, the inhibition of tumor angiogenesis, and the
other, the blockade of HGF-mediated dissociation and invasion of tumor
cells. There are several reports that aggressive angiogenesis in
primary tumors correlates well with the high frequency of distant
metastasis in cancer patients (49
, 50)
. Reduced
angiogenesis by NK4 in the primary tumors may have decreased the
incidence of intravasation of tumor cells, the result being inhibition
of lung metastasis. On the other hand, HGF potently stimulates
dissociation, migration, and invasion of tumor cells (12
, 51, 52, 53)
, and the induction of extracellular protease networks
(such as urokinase-type plasminogen activator and a variety of matrix
metalloproteases) by HGF is involved in invasion and subsequent
metastasis (18
, 53
, 54)
. Indeed, HGF potently stimulated
in-vitro invasion of LLC and Jyg-MC cells through Matrigel
(which mimics basement membrane), whereas NK4 blocked the
HGF-induced invasion of these tumor cells (Fig. 1B)
.
Thus, the blockade of HGF-Met coupling by NK4 might have suppressed the
invasion, motility, and subsequent intravasation of tumor cells,
leading to the inhibition of lung metastasis, in our model.
Cutting off the vascular supply for malignant tumors by an angiogenesis inhibitor is expected to be a promising form of cancer therapy. On the other hand, an angiogenesis inhibitor will not modify the invasive and metastatic phenotype of cancer cells, though angiogenesis-dependent tumor invasion is suppressed by halting angiogenesis in some cases (55) . A number of studies indicated a critical role for HGF in the invasion and metastasis of a wide variety of malignant tumor cells (6, 7, 8, 9, 10) , and inhibition of HGF-Met coupling or signal transduction from the Met has been implicated as a therapeutic strategy to prevent cancer invasion and metastasis (18 , 56, 57, 58) . On the basis of the bifunctional characteristics of NK4 to target both tumor angiogenesis and HGF-mediated invasion, the possibility that NK4 can function as a therapeutic for subjects with cancer warrants ongoing studies.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 This study was supported by a research grant for
science and cancer from the Ministry of Education, Science, Sports, and
Culture of Japan, a research grant for cancer research from the
Ministry of Welfare of Japan, and research grants from the Princess
Takamatsu Cancer Research Fund, the Tokyo Biochemical Research
Foundation, the Foundation for Promotion of Cancer Research in Japan,
Takeda Science Foundation, Nissan Science Foundation, and the Mochida
Memorial Foundation for Medical and Pharmaceutical Research. 
2 To whom requests for reprints should be
addressed, at Division of Biochemistry, Department of Oncology,
Biomedical Research Center, Osaka University Medical School, Suita,
Osaka 565-0871, Japan. Phone: 81-6-6879-3783; Fax: 81-6-6879-3789;
E-mail: nakamura{at}onbich.med.osaka-u.ac.jp
3 The abbreviations used are: HGF, hepatocyte
growth factor; EBM-2, endothelial basal medium; FBS, fetal bovine
serum; MDCK, Madin-Darby canine kidney; CAM, chick chorioallantoic
membrane; bFGF, basic fibroblast growth factor; VEGF, vascular
endothelial growth factor; KDR, human Flk-1; MAPK, mitogen-activated
protein kinase; LLC, Lewis lung carcinoma; Jyg-MC, Jyg-MC(A) murine
mammary carcinoma; PCNA, proliferating cell nuclear antigen; TUNEL,
terminal deoxynucleotidyl transferase-mediated nick end labeling;
HMVEC, human dermal microvascular endothelial cell.
Received 11/30/99. Accepted 10/ 5/00.
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REFERENCES |
|---|
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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vß3 for angiogenesis. Science (Washington DC), 264: 569-571, 1994.This article has been cited by other articles:
|
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  K. Sakai, T. Nakamura, K. Matsumoto, and T. Nakamura Angioinhibitory Action of NK4 Involves Impaired Extracellular Assembly of Fibronectin Mediated by Perlecan-NK4 Association J. Biol. Chem., August 14, 2009; 284(33): 22491 - 22499. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. G. SKOLDENBERG, A. LARSSON, A. JAKOBSON, F. HEDBORG, P. KOGNER, R. H. CHRISTOFFERSON, and F. AZARBAYJANI The Angiogenic Growth Factors HGF and VEGF in Serum and Plasma from Neuroblastoma Patients Anticancer Res, August 1, 2009; 29(8): 3311 - 3319. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Toschi and P. A. Janne Single-Agent and Combination Therapeutic Strategies to Inhibit Hepatocyte Growth Factor/MET Signaling in Cancer Clin. Cancer Res., October 1, 2008; 14(19): 5941 - 5946. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Nie, Z. Shen, J.-B. Wen, O. G.-W. Wong, W. D. Hsueh, L.-F. Huo, H.-F. Kung, B. Jiang, and M. C.M. Lin AAV-HGFK1 and Ad-p53 cocktail therapy prolongs survival of mice with colon cancer Mol. Cancer Ther., September 1, 2008; 7(9): 2855 - 2865. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H.-K. Kim, D.-S. Oh, S.-B. Lee, J.-M. Ha, and Y. A. Joe Antimigratory effect of TK1-2 is mediated in part by interfering with integrin {alpha}2{beta}1 Mol. Cancer Ther., July 1, 2008; 7(7): 2133 - 2141. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Shen, Z. F. Yang, Y. Gao, J. C. Li, H. X. Chen, C. C. Liu, R. T.P. Poon, S. T. Fan, J. M. Luk, K. H. Sze, et al. The Kringle 1 Domain of Hepatocyte Growth Factor Has Antiangiogenic and Antitumor Cell Effects on Hepatocellular Carcinoma Cancer Res., January 15, 2008; 68(2): 404 - 414. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
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|
|
|
|
 |
|
 R. Abounader and J. Laterra Scatter factor/hepatocyte growth factor in brain tumor growth and angiogenesis Neuro-oncol, October 1, 2005; 7(4): 436 - 451. [Abstract] [PDF]
|
|
|
|
 |
|
 T.-L. Cheng, M.-Y. Chang, S.-Y. Huang, C.-C. Sheu, E.-L. Kao, Y.-J. Cheng, and I.-W. Chong Overexpression of Circulating c-Met Messenger RNA Is Significantly Correlated With Nodal Stage and Early Recurrence in Non-Small Cell Lung Cancer Chest, September 1, 2005; 128(3): 1453 - 1460. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. A. Carroll, L. L. Nikitenko, R. Bicknell, and A. L. Harris Antiangiogenic Activity of a Domain Deletion Mutant of Tissue Plasminogen Activator Containing Kringle 2 Arterioscler Thromb Vasc Biol, April 1, 2005; 25(4): 736 - 741. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Sato, N. Maehara, and M. Goggins Gene Expression Profiling of Tumor-Stromal Interactions between Pancreatic Cancer Cells and Stromal Fibroblasts Cancer Res., October 1, 2004; 64(19): 6950 - 6956. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Maemondo, Y. Saijo, K. Narumi, T. Kikuchi, K. Usui, R. Tazawa, K. Matsumoto, T. Nakamura, K. Sasaki, M. Takahashi, et al. Gene Therapy with Secretory Leukoprotease Inhibitor Promoter-Controlled Replication-Competent Adenovirus for Non-Small Cell Lung Cancer Cancer Res., July 1, 2004; 64(13): 4611 - 4620. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Nakano, S. Higashiyama, H. Ohmoto, H. Ishiguro, N. Taniguchi, and Y. Wada The N-terminal Region of NTAK/Neuregulin-2 Isoforms Has an Inhibitory Activity on Angiogenesis J. Biol. Chem., March 19, 2004; 279(12): 11465 - 11470. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Goi, M. Fujioka, Y. Satoh, S. Tabata, K. Koneri, H. Nagano, Y. Hirono, K. Katayama, K. Hirose, and A. Yamaguchi Angiogenesis and Tumor Proliferation/Metastasis of Human Colorectal Cancer Cell Line SW620 Transfected with Endocrine Glands-Derived-Vascular Endothelial Growth Factor, As a New Angiogenic Factor Cancer Res., March 15, 2004; 64(6): 1906 - 1910. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Yoshida, T. Harada, M. Mitsunari, T. Iwabe, Y. Sakamoto, S. Tsukihara, Y. Iba, S. Horie, and N. Terakawa Hepatocyte Growth Factor/Met System Promotes Endometrial and Endometriotic Stromal Cell Invasion via Autocrine and Paracrine Pathways J. Clin. Endocrinol. Metab., February 1, 2004; 89(2): 823 - 832. [Abstract] [Full Text] [PDF]
|
|
|
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M. Krix, F. Kiessling, S. Vosseler, N. Farhan, M. M. Mueller, P. Bohlen, N. E. Fusenig, and S. Delorme Sensitive Noninvasive Monitoring of Tumor Perfusion during Antiangiogenic Therapy by Intermittent Bolus-Contrast Power Doppler Sonography Cancer Res., December 1, 2003; 63(23): 8264 - 8270. [Abstract] [Full Text] [PDF]
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S. J. Kim, M. Johnson, K. Koterba, M. H. Herynk, H. Uehara, and G. E. Gallick Reduced c-Met Expression by an Adenovirus Expressing a c-Met Ribozyme Inhibits Tumorigenic Growth and Lymph Node Metastases of PC3-LN4 Prostate Tumor Cells in an Orthotopic Nude Mouse Model Clin. Cancer Res., November 1, 2003; 9(14): 5161 - 5170. [Abstract] [Full Text] [PDF]
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X. Wang, P. Le, C. Liang, J. Chan, D. Kiewlich, T. Miller, D. Harris, L. Sun, A. Rice, S. Vasile, et al. Potent and selective inhibitors of the Met [hepatocyte growth factor/scatter factor (HGF/SF) receptor] tyrosine kinase block HGF/SF-induced tumor cell growth and invasion Mol. Cancer Ther., November 1, 2003; 2(11): 1085 - 1092. [Abstract] [Full Text] [PDF]
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 Y.-W. Zhang, Y. Su, O. V. Volpert, and G. F. V. Woude Hepatocyte growth factor/scatter factor mediates angiogenesis through positive VEGF and negative thrombospondin 1 regulation PNAS, October 28, 2003; 100(22): 12718 - 12723. [Abstract] [Full Text] [PDF]
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M. A. Brockmann, A. Papadimitriou, M. Brandt, R. Fillbrandt, M. Westphal, and K. Lamszus Inhibition of Intracerebral Glioblastoma Growth by Local Treatment with the Scatter Factor/Hepatocyte Growth Factor-Antagonist NK4 Clin. Cancer Res., October 1, 2003; 9(12): 4578 - 4585. [Abstract] [Full Text] [PDF]
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T. Merkulova-Rainon, P. England, S. Ding, C. Demerens, and G. Tobelem The N-terminal Domain of Hepatocyte Growth Factor Inhibits the Angiogenic Behavior of Endothelial Cells Independently from Binding to the c-met Receptor J. Biol. Chem., September 26, 2003; 278(39): 37400 - 37408. [Abstract] [Full Text] [PDF]
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T. Manabe, K. Mizumoto, E. Nagai, K. Matsumoto, T. Nakamura, T. Nukiwa, M. Tanaka, and T. Matsuda Cell-based Protein Delivery System for the Inhibition of the Growth of Pancreatic Cancer: NK4 Gene-transduced Oral Mucosal Epithelial Cell Sheet Clin. Cancer Res., August 1, 2003; 9(8): 3158 - 3166. [Abstract] [Full Text] [PDF]
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M. Numasaki, J.-i. Fukushi, M. Ono, S. K. Narula, P. J. Zavodny, T. Kudo, P. D. Robbins, H. Tahara, and M. T. Lotze Interleukin-17 promotes angiogenesis and tumor growth Blood, April 1, 2003; 101(7): 2620 - 2627. [Abstract] [Full Text] [PDF]
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K. S. Kim, Y.-K. Hong, Y. A. Joe, Y. Lee, J.-Y. Shin, H.-E. Park, I.-H. Lee, S.-Y. Lee, D.-K. Kang, S.-I. Chang, et al. Anti-angiogenic Activity of the Recombinant Kringle Domain of Urokinase and Its Specific Entry into Endothelial Cells J. Biol. Chem., March 21, 2003; 278(13): 11449 - 11456. [Abstract] [Full Text] [PDF]
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N. Wajih and D. C. Sane Angiostatin selectively inhibits signaling by hepatocyte growth factor in endothelial and smooth muscle cells Blood, March 1, 2003; 101(5): 1857 - 1863. [Abstract] [Full Text] [PDF]
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T. Kikuchi, M. Maemondo, K. Narumi, K. Matsumoto, T. Nakamura, and T. Nukiwa Tumor suppression induced by intratumor administration of adenovirus vector expressing NK4, a 4-kringle antagonist of hepatocyte growth factor, and naive dendritic cells Blood, December 1, 2002; 100(12): 3950 - 3959. [Abstract] [Full Text] [PDF]
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M. Saimura, E. Nagai, K. Mizumoto, N. Maehara, Y. A. Minamishima, M. Katano, K. Matsumoto, T. Nakamura, and M. Tanaka Tumor Suppression through Angiogenesis Inhibition by SUIT-2 Pancreatic Cancer Cells Genetically Engineered to Secrete NK4 Clin. Cancer Res., October 1, 2002; 8(10): 3243 - 3249. [Abstract] [Full Text] [PDF]
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 Y. To, M. Dohi, K. Matsumoto, R. Tanaka, A. Sato, K. Nakagome, T. Nakamura, and K. Yamamoto A Two-way Interaction between Hepatocyte Growth Factor and Interleukin-6 in Tissue Invasion of Lung Cancer Cell Line Am. J. Respir. Cell Mol. Biol., August 1, 2002; 27(2): 220 - 226. [Abstract] [Full Text] [PDF]
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 Y. Saijo, M. Tanaka, M. Miki, K. Usui, T. Suzuki, M. Maemondo, X. Hong, R. Tazawa, T. Kikuchi, K. Matsushima, et al. Proinflammatory Cytokine IL-1{beta} Promotes Tumor Growth of Lewis Lung Carcinoma by Induction of Angiogenic Factors: In Vivo Analysis of Tumor-Stromal Interaction J. Immunol., July 1, 2002; 169(1): 469 - 475. [Abstract] [Full Text] [PDF]
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D. Tomioka, N. Maehara, K. Kuba, K. Mizumoto, M. Tanaka, K. Matsumoto, and T. Nakamura Inhibition of Growth, Invasion, and Metastasis of Human Pancreatic Carcinoma Cells by NK4 in an Orthotopic Mouse Model Cancer Res., October 1, 2001; 61(20): 7518 - 7524. [Abstract] [Full Text] [PDF]
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 K. Matsumoto, H. Yoshitomi, J. Rossant, and K. S. Zaret Liver Organogenesis Promoted by Endothelial Cells Prior to Vascular Function Science, October 19, 2001; 294(5542): 559 - 563. [Abstract] [Full Text] [PDF]
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