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Tumor Biology |
Department of Biophysics, Institute for Cancer Research, The Norwegian Radium Hospital, N-0310 Oslo, Norway
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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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Melanoma cells have been shown to produce and secrete a wide variety of angiogenic factors, including VEGF3 (13, 14, 15, 16) , IL-8 (17, 18, 19) , PD-ECGF (20 , 21) , and bFGF (22, 23, 24) . VEGF, also known as vascular permeability factor, is a strong specific mitogen for endothelial cells and may also stimulate endothelial cell migration and reorganization (25 , 26) . IL-8, which belongs to the superfamily of CXC chemokines, is a multifunctional cytokine that exhibits potent angiogenic activities both in vitro and in vivo (27 , 28) . PD-ECGF, also known as thymidine phosphorylase and gliostatin, stimulates endothelial cell mitogenesis and chemotaxis in vitro and is strongly angiogenic in vivo, possibly through modulation of nucleotide metabolism (29) . bFGF, which belongs to the family of heparin-binding growth factors, is a multifunctional protein having a well-established key role in tumor angiogenesis (30, 31, 32) .
There is significant experimental evidence that these angiogenic factors are involved in the growth and metastasis of malignant melanoma. Inoculation of human melanoma cells transfected with the gene encoding VEGF into immune-deficient mice results in tumors with increased vascularization (13, 14, 15) , microvessel permeability (14 , 15 , 33) , and volumetric growth rate (14 , 33) . The constitutive expression of IL-8 in human melanoma cells is correlated with their lung colonization efficiency after i.v. inoculation in immune-deficient mice (17) . Human melanoma cells show increased lung colonization efficiency after transfection with the gene encoding VEGF (15) or IL-8 (18) . Moreover, xenografted tumors initiated from human melanoma cell lines transfected with antisense-VEGF (14 , 15) or antisense-bFGF (24) show reduced vascularization and growth.
A panel of angiogenic factors may thus be involved in the angiogenesis, growth, and metastasis of melanomas, and the panel may differ among tumors differing in angiogenic activity (34, 35, 36) . However, angiogenic factors that show sufficiently high constitutive expression to promote spontaneous metastasis have not been identified conclusively thus far. The work reported here was aimed at investigating whether the angiogenesis and metastasis of melanoma may be promoted by VEGF, IL-8, PD-ECGF, and/or bFGF. Four human melanoma xenograft lines showing widely different constitutive expression of these angiogenic factors were included in the study. The specific role of each of the factors was investigated by assessing tumor angiogenesis and metastasis in mice treated with neutralizing antibody.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Cell Lines.
Four human melanoma cell lines (A-07, D-12, R-18, and U-25) were
included in the study (37)
. They were maintained in
monolayer culture in RPMI 1640 (25 mM HEPES and
L-glutamine) supplemented with 13% bovine calf serum, 250
mg/l penicillin, and 50 mg/l streptomycin. The cultures were incubated
at 37°C in a humidified atmosphere of 5% CO2
in air and subcultured twice a week. The cell lines were verified to be
free from Mycoplasma contamination.
ELISA.
Commercial ELISA kits (Quantikine; R&D Systems, Abingdon, United
Kingdom) were used according to the instructions of the manufacturer to
measure VEGF, IL-8, and bFGF concentrations in culture medium. Medium
samples from cultures in exponential growth were collected 24 h
after change of medium. Cell numbers at the time of medium change and
at the time of medium collection were determined by using a
hemocytometer and a phase contrast microscope. The medium samples were
centrifuged to remove particles, diluted to appropriate concentrations,
and analyzed in duplicate at several dilutions. Absorbances were
determined at 450 nm. Readings at 570 nm were subtracted from the
readings at 450 nm to correct for optical imperfections. A standard
curve was obtained by linear regression analysis of protein
concentration versus absorbance in a double logarithmic
plot. The rate of protein secretion
(Rsec) was calculated as:
 |
C is the increase in protein concentration
during the time interval t. Here, t was
24 h. Ni and
Nf are the initial and final cell
numbers, and V is the volume of medium. Replicate cultures
were used to determine Ni. The second
factor of the product is based on the assumption that the cell number
increased exponentially with time during t, an assumption
that was verified to be valid.
Western Blotting.
Cells from cultures in exponential growth were washed in PBS and boiled
in Laemmli lysis buffer for 5 min (38)
. Proteins were
separated by SDS-PAGE and transferred to a polyvinylidene fluoride
membrane. Membranes were incubated with antihuman VEGF rabbit
polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA),
antihuman IL-8 mouse monoclonal antibody (R&D Systems), antihuman
PD-ECGF goat polyclonal antibody (R&D Systems), or antihuman bFGF
rabbit polyclonal antibody (Oncogene Science, Cambridge, MA) for 60
min. Bound antibody was detected by using a biotin-streptavidin
alkaline phosphatase staining procedure. Recombinant human VEGF, IL-8,
PD-ECGF, or bFGF (R&D Systems) was used as positive control. The
specificity of the antibody-antigen interactions was confirmed by
peptide competition studies and by incubation of membranes in solutions
without primary antibody. Protein molecular weights were estimated by
using broad range or high range prestained standards according to the
instructions of the manufacturer (SDS-PAGE standards; Bio-Rad
Laboratories, Hercules, CA).
Immunohistochemistry.
Immunohistochemical staining of tumor tissue was performed by using an
indirect immunoperoxidase method. Tumors were fixed in
phosphate-buffered 4% paraformaldehyde or snap-frozen in liquid
nitrogen. Antihuman VEGF rabbit polyclonal antibody (Santa Cruz
Biotechnology), antihuman IL-8 rabbit polyclonal antibody (Endogen,
Woburn, MA), antihuman PD-ECGF goat polyclonal antibody (R&D Systems),
or antihuman bFGF rabbit polyclonal antibody (Santa Cruz Biotechnology)
was used as primary antibody. Controls included omission of the primary
antibody, incubation with normal rabbit immunoglobulin or normal rabbit
serum, and incubation with blocking peptides before staining. The
sections were counterstained with hematoxylin.
Angiogenesis Assay.
Tumor angiogenesis was assessed by using an intradermal angiogenesis
assay (39)
. A 100-µl Hamilton syringe was used to
inoculate aliquots of 10 µl of cell suspension into the flanks of
mice. The inoculation point lay above the s.c. muscle tissue in the
deeper part of the dermis. The number of cells per inoculum was
1.0 x 106. The mice were killed
on day 7 after the inoculation. Small vascularized tumors had developed
in the inoculation sites at that time. The skin around the inoculation
sites was removed, and the tumors were located with a dissecting
microscope. The capillaries in the dermis oriented toward the tumors
were counted, and the diameters of the tumors were measured, using
an ocular with a scale. The number of capillaries was corrected for the
background, determined after the injection of 10 µl of HBSS.
Angiogenesis was quantified as number of capillaries per tumor or
number of capillaries per mm of tumor circumference.
Lung Colonization Assay.
Aliquots of 3.0 x 106 A-07 cells
or 3.0 x 105 D-12 cells suspended
in 0.2 ml of HBSS were inoculated into the lateral tail vein of mice by
using a tuberculin syringe with a 26-gauge needle. The mice were killed
and autopsied 5 weeks after the inoculation. The lungs were removed,
rinsed in HBSS, and fixed in Bouin’s solution for 24 h to
facilitate the scoring of colonies. The number of surface colonies was
determined by using a stereomicroscope.
Spontaneous Metastasis Assay.
Aliquots of 3.5 x 105 A-07 or
D-12 cells suspended in 10 µl of HBSS were inoculated intradermally
in the left flank of mice, using the same procedure as used in the
angiogenesis assay described above. The inoculations were performed
24 h after the mice had been immunosuppressed by 5.0 Gy of
whole-body irradiation. The whole-body irradiation was performed by
using a Siemens Stabilipan X-ray unit, operated at 220 kV, 19–20 mA,
and with 0.5-mm copper filtration (40)
. The tumors were
removed surgically when the largest diameter had attained 
10 mm, and
the wounds were closed with surgical clips. The mice were examined for
clinical signs of metastases twice a week. They were killed and
autopsied 3 months after the primary tumor was removed or when they
were moribund. The lungs were examined for the presence of macroscopic
metastases as described above.
Treatment with Neutralizing Antibody in Vivo.
The specific roles of VEGF, IL-8, PD-ECGF, and bFGF in tumor
angiogenesis, lung colonization, and spontaneous metastasis were
investigated by treating host mice with neutralizing antibody against
these angiogenic factors. The antibodies used for treatment were
antihuman VEGF mouse monoclonal antibody (R&D Systems), antihuman IL-8
mouse monoclonal antibody (R&D Systems), antihuman PD-ECGF goat
polyclonal antibody (R&D Systems), and antihuman bFGF goat polyclonal
antibody (R&D Systems). The antibody solutions were diluted in PBS and
administered to the mice in volumes of 0.25 ml by i.p. injection. In
the angiogenesis and lung colonization experiments, the treatments
consisted of four doses of 25 µg (VEGF and bFGF) or 100 µg (IL-8
and PD-ECGF) of antibody given in intervals of 24 h. The first
dose was given 1 h before the tumor cell inoculation. In the
spontaneous metastasis experiments, the treatments consisted of eight
doses of antibody given daily the last 8 days before the primary tumor
was removed.
Treatment with Neutralizing Antibody in Vitro.
Possible cytotoxic or antiproliferative effects of the neutralizing
antibodies described above were investigated in vitro. A-07,
D-12, R-18, or U-25 cells were cultured in RPMI 1640 (25
mM HEPES and L-glutamine)
supplemented with 13% bovine calf serum, 250 mg/l penicillin, and 50
mg/l streptomycin in the absence or presence of 5 µg/ml of antibody
for up to 8 days. The number of cells in the cultures was determined 2,
4, 6, or 8 days after the cultures were initiated by counting cells in
a hemocytometer.
Statistical Analysis.
Results are presented as arithmetic mean ± SE.
Statistical comparisons of data were performed by one-way ANOVA. When
significant differences were found, the Dunnett’s method was used to
identify the data sets that differed from the control data. Probability
values of P < 0.05 were considered
significant. All Ps were determined from two-sided tests.
The statistical analysis was performed by using SigmaStat statistical
software (Jandel Scientific GmbH, Erkrath, Germany).
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RESULTS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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. A-07 showed higher VEGF secretion than U-25 (P < 0.05), which in turn showed higher VEGF secretion than D-12 and
R-18 (P < 0.05). The IL-8 secretion was also
higher in A-07 than in the other lines (P < 0.05). Moreover, D-12 showed higher IL-8 secretion than U-25
(P < 0.05), which in turn showed higher IL-8
secretion than R-18 (P < 0.05).
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. A-07 showed substantially higher VEGF, IL-8, and bFGF expression than
the other lines. Moreover, six specific bFGF bands were seen in A-07,
whereas D-12, R-18, and U-25 showed only 2–4 specific bFGF bands.
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. Also D-12, R-18, and U-25 stained positive for VEGF, IL-8, and bFGF
in vivo, but the staining was weaker than that in A-07
tumors for all proteins. Significant expression of PD-ECGF could not be
detected in D-12, R-18, or U-25 tumors.
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. The angiogenic response, measured as number of tumor-oriented
capillaries per tumor at day 7 after the inoculation of 1.0 x 106 cells, differed substantially among
the lines. A-07 evoked the strongest angiogenic response, followed by
D-12, R-18, and U-25. The differences in number of tumor-oriented
capillaries between A-07 and D-12, D-12 and R-18, and R-18 and U-25
were all statistically significant (P < 0.05). The angiogenic response evoked by A-07 was inhibited in mice
treated with neutralizing antibody against VEGF, IL-8, PD-ECGF, or bFGF
(P < 0.05). Treatment with neutralizing
antibody against VEGF or IL-8 also reduced the angiogenic response
evoked by D-12 (P < 0.05) but not treatment
with neutralizing antibody against PD-ECGF or bFGF
(P > 0.05). The angiogenic response evoked
by R-18 and U-25 was inhibited in mice treated with neutralizing
antibody against VEGF (P < 0.05) but not in
mice treated with neutralizing antibody against IL-8, PD-ECGF, or bFGF
(P > 0.05). Similar results and identical
conclusions were achieved by expressing angiogenic response as number
of tumor-oriented capillaries per mm of tumor circumference (data not
shown).
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. The lung colonization efficiency of A-07 was reduced in mice treated
with neutralizing antibody against VEGF, IL-8, PD-ECGF, or bFGF
(P < 0.05). Treatment with neutralizing
antibody against VEGF or IL-8 also reduced the lung colonization
efficiency of D-12 (P < 0.05) but not
treatment with neutralizing antibody against PD-ECGF or bFGF
(P > 0.05). R-18 and U-25 cells do not
develop lung colonies after i.v. inoculation in mice (41)
.
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50% of the mice (Fig. 5)
. The metastatic efficiency of A-07 tumors was reduced in mice treated
with neutralizing antibody against VEGF, IL-8, PD-ECGF, or bFGF
(P < 0.05). Treatment with neutralizing
antibody against VEGF or IL-8 also reduced the metastatic efficiency of
D-12 tumors (P < 0.05) but not treatment
with neutralizing antibody against PD-ECGF or bFGF
(P > 0.05).
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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The differences among A-07, D-12, R-18, and U-25 cells in the expression of VEGF and IL-8 were more pronounced when the expression was measured by ELISA than when measured by Western blotting. Western blot analysis gives information on intracellular protein concentrations, whereas ELISA analysis provides information on the rate of protein secretion. There is no a priori reason to believe that the rate of protein secretion should be correlated to the intracellular concentration of protein. In fact, the present study suggests that the rate of protein secretion cannot be predicted from the intracellular protein concentration as determined by Western blot analysis. The rate of protein secretion is probably more relevant for the rate of tumor neovascularization than is the intracellular concentration of protein, because angiogenesis is induced by binding of protein to endothelial cell receptors.
A-07 cells, which evoked the strongest angiogenic response in vivo, showed substantially higher expression of bFGF than did D-12, R-18, and U-25 cells. bFGF may promote tumor angiogenesis directly by acting synergistically with VEGF (42, 43, 44) and indirectly by up-regulating the synthesis and secretion of VEGF (45 , 46) .
The expression of VEGF, IL-8, PD-ECGF, and bFGF of A-07, D-12, R-18,
and U-25 tumors, determined by immunohistochemical analysis, was in
agreement with that of the corresponding cell lines in
vitro, i.e., A-07 tumors showed substantially stronger
staining for VEGF, IL-8, and bFGF than did D-12, R-18, and
U-25 tumors and were the only tumors that stained positive for PD-ECGF,
consistent with the in vitro data in Table 1
and Fig. 1
.
This observation was not expected, because A-07, D-12, R-18, and U-25
tumors differ substantially in the fraction of hypoxic cells
(47)
, and hypoxia may up-regulate the expression of
VEGF (48
, 49)
, IL-8 (19
, 50)
, and PD-ECGF
(51)
. The expression of VEGF has been shown to be
up-regulated in A-07, D-12, R-18, and U-25 cells exposed to hypoxia
in vitro (16
, 52)
. However, hypoxia-induced
up-regulation of VEGF, IL-8, or PD-ECGF was not detected in the
immunohistochemical analysis. All tissue sections stained homogeneously
for VEGF, IL-8, and PD-ECGF, i.e., the staining was not more
intense in regions adjacent to necrosis than in regions close to
functional capillaries, possibly because VEGF, IL-8, and PD-ECGF were
secreted and diffused from hypoxic tissue into surrounding normoxic
tissue. The staining patterns of VEGF, IL-8, and PD-ECGF differed
substantially from the staining pattern of the hypoxia marker
pimonidazole in A-07, D-12, R-18, and U-25 tumors
(53)
.
The angiogenic factors expressed in vivo were produced primarily by the parenchymal melanoma cells and were thus of human origin. The immunohistochemical analysis showed that the angiogenic factors were mainly localized intracellularly in melanoma cells and not in stromal host cells. Moreover, host cells, i.e., macrophages, leukocytes, and fibroblasts, have been shown to constitute <1% of the total number of cells in A-07, D-12, R-18, and U-25 tumors (37) . Our conclusion is also supported by the observation that the expression of angiogenic factors in vivo was similar to that in vitro.
The angiogenic response after intradermal cell inoculation differed significantly among the melanoma lines. The sequence of the lines from high to low angiogenic response, measured as number of tumor-oriented capillaries per tumor or number of tumor-oriented capillaries per mm of tumor circumference, was A-07 > D-12 > R-18 > U-25. The tumor growth rate has also been shown to differ significantly among the melanoma lines (39) . The sequence of the lines from high to low volumetric growth rate is identical to that for angiogenic response, i.e., A-07 > D-12 > R-18 > U-25. The close association between volumetric growth rate and angiogenic response suggests that the growth of A-07, D-12, R-18, and U-25 tumors is governed primarily by the rate of angiogenesis.
The angiogenic factors promoting angiogenesis in A-07, D-12, R-18, and U-25 tumors, identified by measuring tumor-induced angiogenic response in mice treated with neutralizing antibody, differed among the lines. The angiogenesis of A-07 tumors, which showed the highest rate of angiogenesis, was promoted by VEGF, IL-8, and PD-ECGF, as well as bFGF. VEGF and IL-8, but probably not PD-ECGF or bFGF, promoted angiogenesis in D-12 tumors, which showed the second highest rate of angiogenesis. The angiogenesis of R-18 and U-25 tumors was promoted by VEGF but probably not by IL-8, PD-ECGF, or bFGF. All lines expressed bFGF, whereas A-07 was the only line secreting bFGF, suggesting that promotion of tumor angiogenesis by bFGF may require bFGF secretion. A-07 and D-12 showed significantly higher secretion of IL-8 than did R-18 and U-25, indicating that promotion of tumor angiogenesis by IL-8 may require high IL-8 concentrations. Our observations suggest that multiple angiogenic factors are involved in the angiogenesis of highly angiogenic melanomas, whereas the angiogenesis in poorly angiogenic melanomas may be promoted solely by VEGF. Angiogenic factors other than those studied here may also be involved in the angiogenesis of melanoma, including angiogenin (54) and angiopoietin-1 (55) .
The angiogenic factors that were found to promote tumor angiogenesis were also found to be essential for tumor metastasis. Lung colonization and spontaneous metastasis in A-07 were inhibited by treatment with neutralizing antibody against VEGF, IL-8, PD-ECGF, or bFGF, as was A-07-induced angiogenesis. Treatment with neutralizing antibody against VEGF or IL-8 inhibited lung colonization and spontaneous metastasis as well as angiogenesis in D-12, whereas none of these biological phenomena was inhibited by treatment with neutralizing antibody against PD-ECGF or bFGF. These observations suggest that spontaneous lung metastasis in A-07 and D-12 tumors is governed by the angiogenic potential of the tumor cells and hence controlled by the angiogenic factors controlling the angiogenesis.
The metastatic process is composed of a cascade of linked, sequential,
and highly selective steps involving multiple host-tumor interactions.
These steps include invasion of tumor cells into blood vessels,
survival in the blood circulation, arrest in the capillary bed of a
secondary organ, extravasation into the secondary organ interstitium
and parenchyma, and tumor cell proliferation and angiogenesis in the
secondary organ. The metastatic propensity of a tumor may be influenced
by the angiogenic potential of the tumor cells by two independent
mechanisms: high microvessel density in the primary tumor may enhance
the opportunity of tumor cells to gain access to the blood circulation;
and elevated capacity to induce neovascularization may increase the
probability of tumor cells trapped in secondary organ capillary beds to
give rise to macroscopic tumor growth (56)
. The
probability of spontaneous lung metastasis in A-07 and D-12 tumors was
probably limited by the capacity of the tumor cells to induce
angiogenesis in the lungs, because spontaneous lung metastasis and lung
colonization were inhibited by treatment with neutralizing antibody
against the same angiogenic factors. However, it cannot be excluded
that the probability of spontaneous lung metastasis was influenced by
the angiogenic potential of the tumor cells also via the microvessel
density of the primary tumor. Interestingly, the microvessel density is
2.5-fold higher in A-07 tumors than in D-12 tumors
(57)
.
The antibody treatments that inhibited lung colonization and
spontaneous metastasis in A-07 and D-12 melanomas lasted 4 and 8 days,
respectively. The reduced lung colonization and spontaneous metastasis
in antibody-treated mice were most likely a consequence of inhibited
angiogenesis rather than of cytotoxic or antiproliferative effects of
the antibodies, because the growth of melanoma cells in
vitro was not inhibited by antibody treatment. This interpretation
is based on the assumption that neovascularization and macroscopic
growth of lung colonies can be inhibited by treatment with neutralizing
antibodies against angiogenic factors also when the antibody treatment
is given to lung colonies consisting of less than 50 cells, which is
the maximum size of 4-day-old A-07 and D-12 lung colonies. This
assumption is not consistent with the general current thinking that
angiogenesis is initiated when the tumor mass reaches a diameter of
1 mm. However, the validity of our assumption is strongly supported
by recent observations by Li et al. (58)
studying the initial phases of tumor cell-induced angiogenesis in skin
window chambers. They observed modifications of the host vasculature
when the tumor mass reached 60–80 cells and saw functional new blood
vessels when the tumor mass reached 100–300 cells. Moreover, they
found that ex-flk 1, a truncated soluble VEGF receptor protein known to
be antiangiogenic and nontoxic, prevented angiogenesis and tumor growth
in four of six window chambers when administered as a single dose at
the same time as 40–50 tumor cells were inoculated into the window
chambers. This observation led the authors to conclude that individual
tumor cells are dependent on early angiogenic activities for both
survival and proliferation in vivo (58)
.
Previous studies have shown that lung colonization of human melanoma cells in immune-deficient mice can be inhibited by treatment with neutralizing antibody against VEGF (52) or by treatment with agents blocking the receptor of IL-8 (59) . The present study confirms these observations and shows further for the first time that treatment with neutralizing antibody against VEGF or IL-8 can inhibit spontaneous lung metastasis of human melanoma xenografts. It has been shown by others that treatment with neutralizing antibody against VEGF can inhibit spontaneous metastasis of human epidermoid carcinoma xenografts (60) , human prostate carcinoma xenografts (61) , and human colon and gastric carcinoma xenografts (62) , and treatment with neutralizing antibody against IL-8 can inhibit spontaneous metastasis of non-small cell lung carcinoma xenografts (63) . Moreover, the study reported here also shows for the first time that spontaneous lung metastasis of human melanoma xenografts can be inhibited by treatment with neutralizing antibody against PD-ECGF or bFGF.
The most important finding reported here is that the angiogenesis and spontaneous lung metastasis of A-07 melanoma xenografts can be inhibited by treatment with neutralizing antibody against VEGF, IL-8, PD-ECGF, or bFGF. Each of these angiogenic factors was probably essential for the angiogenesis and metastasis of A-07 tumors, because inhibition of one of them could not be compensated for by the others. This observation suggests that inhibition of a single angiogenic factor pathway may be beneficial in the treatment of malignant melanoma. However, it is unlikely that complete inhibition of tumor growth and metastasis of highly angiogenic melanomas would be possible by this strategy, because multiple angiogenic factors are involved. Efficient antiangiogenic treatment of malignant melanoma may require identification and blocking of common functional features of several angiogenic factors. Treatment strategies that target the common signaling cascades in the end organ of angiogenesis, i.e., the endothelial cell, may be more universally effective than those targeting specific angiogenic factors or their receptors.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported by grants from The Norwegian Cancer
Society. 
2 To whom requests for reprints should be
addressed, at Department of Biophysics, Institute for Cancer Research,
The Norwegian Radium Hospital, Montebello, N-0310 Oslo, Norway. Phone:
47-2293-4279; Fax: 47-2293-4270; E-mail: e.k.rofstad{at}labmed.uio.no
3 The abbreviations used are: VEGF, vascular
endothelial growth factor; bFGF, basic fibroblast growth factor; IL,
interleukin; PD-ECGF, platelet-derived endothelial cell growth
factor.
Received 12/29/99. Accepted 7/ 6/00.
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REFERENCES |
|---|
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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chemokines inhibits the tumour growth and pulmonary metastasis of human melanoma cells in nude mice. Melanoma Res., 9: 105-114, 1999.[Medline]
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 M. Galie, C. Sorrentino, M. Montani, L. Micossi, E. Di Carlo, T. D'Antuono, L. Calderan, P. Marzola, D. Benati, F. Merigo, et al. Mammary carcinoma provides highly tumourigenic and invasive reactive stromal cells Carcinogenesis, November 1, 2005; 26(11): 1868 - 1878. [Abstract] [Full Text] [PDF]
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 M. Rusnati, M. Camozzi, E. Moroni, B. Bottazzi, G. Peri, S. Indraccolo, A. Amadori, A. Mantovani, and M. Presta Selective recognition of fibroblast growth factor-2 by the long pentraxin PTX3 inhibits angiogenesis Blood, July 1, 2004; 104(1): 92 - 99. [Abstract] [Full Text] [PDF]
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B. Rousseau, F. Larrieu-Lahargue, S. Javerzat, F. Guilhem-Ducleon, F. Beermann, and A. Bikfalvi The tyrp1-Tag/tyrp1-FGFR1-DN Bigenic Mouse: A Model for Selective Inhibition of Tumor Development, Angiogenesis, and Invasion into the Neural Tissue by Blockade of Fibroblast Growth Factor Receptor Activity Cancer Res., April 1, 2004; 64(7): 2490 - 2495. [Abstract] [Full Text] [PDF]
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 R. Abe, T. Shimizu, S.-i. Yamagishi, A. Shibaki, S. Amano, Y. Inagaki, H. Watanabe, H. Sugawara, H. Nakamura, M. Takeuchi, et al. Overexpression of Pigment Epithelium-Derived Factor Decreases Angiogenesis and Inhibits the Growth of Human Malignant Melanoma Cells in Vivo Am. J. Pathol., April 1, 2004; 164(4): 1225 - 1232. [Abstract] [Full Text] [PDF]
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H. Takeuchi, A. Fujimoto, M. Tanaka, T. Yamano, E. Hsueh, and D. S. B. Hoon CCL21 Chemokine Regulates Chemokine Receptor CCR7 Bearing Malignant Melanoma Cells Clin. Cancer Res., April 1, 2004; 10(7): 2351 - 2358. [Abstract] [Full Text] [PDF]
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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]
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E. K. Rofstad, B. Mathiesen, and K. Galappathi Increased Metastatic Dissemination in Human Melanoma Xenografts after Subcurative Radiation Treatment: Radiation-induced Increase in Fraction of Hypoxic Cells and Hypoxia-induced Up-Regulation of Urokinase-Type Plasminogen Activator Receptor Cancer Res., January 1, 2004; 64(1): 13 - 18. [Abstract] [Full Text] [PDF]
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L. P. Lefter, M. Sunamura, T. Furukawa, K. Takeda, N. Kotobuki, M. Oshimura, S. Matsuno, and A. Horii Inserting Chromosome 18 into Pancreatic Cancer Cells Switches Them to a Dormant Metastatic Phenotype Clin. Cancer Res., October 15, 2003; 9(13): 5044 - 5052. [Abstract] [Full Text] [PDF]
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 N. Patel, L. Sun, D. Moshinsky, H. Chen, K. M. Leahy, P. Le, K. G. Moss, X. Wang, A. Rice, D. Tam, et al. A Selective and Oral Small Molecule Inhibitor of Vascular Epithelial Growth Factor Receptor (VEGFR)-2 and VEGFR-1 Inhibits Neovascularization and Vascular Permeability J. Pharmacol. Exp. Ther., September 1, 2003; 306(3): 838 - 845. [Abstract] [Full Text] [PDF]
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 D. C. Lev, M. Ruiz, L. Mills, E. C. McGary, J. E. Price, and M. Bar-Eli Dacarbazine Causes Transcriptional Up-Regulation of Interleukin 8 and Vascular Endothelial Growth Factor in Melanoma Cells: A Possible Escape Mechanism from Chemotherapy Mol. Cancer Ther., August 1, 2003; 2(8): 753 - 763. [Abstract] [Full Text] [PDF]
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E. K. Rofstad, K. Henriksen, K. Galappathi, and B. Mathiesen Antiangiogenic Treatment with Thrombospondin-1 Enhances Primary Tumor Radiation Response and Prevents Growth of Dormant Pulmonary Micrometastases after Curative Radiation Therapy in Human Melanoma Xenografts Cancer Res., July 15, 2003; 63(14): 4055 - 4061. [Abstract] [Full Text] [PDF]
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 I. Feoktistov, S. Ryzhov, A. E. Goldstein, and I. Biaggioni Mast Cell-Mediated Stimulation of Angiogenesis: Cooperative Interaction Between A2B and A3 Adenosine Receptors Circ. Res., March 21, 2003; 92(5): 485 - 492. [Abstract] [Full Text] [PDF]
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O. Straume, D. G. Jackson, and L. A. Akslen Independent Prognostic Impact of Lymphatic Vessel Density and Presence of Low-Grade Lymphangiogenesis in Cutaneous Melanoma Clin. Cancer Res., January 1, 2003; 9(1): 250 - 256. [Abstract] [Full Text] [PDF]
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 Y. Mishima, Y. Matsumoto-Mishima, Y. Terui, M. Katsuyama, M. Yamada, M. Mori, Y. Ishizaka, K. Ikeda, J.-i. Watanabe, N. Mizunuma, et al. Leukemic Cell-Surface CD13/Aminopeptidase N and Resistance to Apoptosis Mediated by Endothelial Cells J Natl Cancer Inst, July 3, 2002; 94(13): 1020 - 1028. [Abstract] [Full Text] [PDF]
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I. Feoktistov, A. E. Goldstein, S. Ryzhov, D. Zeng, L. Belardinelli, T. Voyno-Yasenetskaya, and I. Biaggioni Differential Expression of Adenosine Receptors in Human Endothelial Cells: Role of A2B Receptors in Angiogenic Factor Regulation Circ. Res., March 22, 2002; 90(5): 531 - 538. [Abstract] [Full Text] [PDF]
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F. De Marchis, D. Ribatti, C. Giampietri, A. Lentini, D. Faraone, M. Scoccianti, M. C. Capogrossi, and A. Facchiano Platelet-derived growth factor inhibits basic fibroblast growth factor angiogenic properties in vitro and in vivo through its alpha receptor Blood, March 15, 2002; 99(6): 2045 - 2053. [Abstract] [Full Text] [PDF]
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E. K. Rofstad, H. Rasmussen, K. Galappathi, B. Mathiesen, K. Nilsen, and B. A. Graff Hypoxia Promotes Lymph Node Metastasis in Human Melanoma Xenografts by Up-Regulating the Urokinase-Type Plasminogen Activator Receptor Cancer Res., March 1, 2002; 62(6): 1847 - 1853. [Abstract] [Full Text] [PDF]
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O. Straume and L. A. Akslen Importance of Vascular Phenotype by Basic Fibroblast Growth Factor, and Influence of the Angiogenic Factors Basic Fibroblast Growth Factor/Fibroblast Growth Factor Receptor-1 and Ephrin-A1/EphA2 on Melanoma Progression Am. J. Pathol., March 1, 2002; 160(3): 1009 - 1019. [Abstract] [Full Text] [PDF]
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 K. Satyamoorthy, G. Li, B. Vaidya, J. Kalabis, and M. Herlyn Insulin-like Growth Factor-I-induced Migration of Melanoma Cells Is Mediated by Interleukin-8 Induction Cell Growth Differ., February 1, 2002; 13(2): 87 - 93. [Abstract] [Full Text] [PDF]
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C. Berking, R. Takemoto, H. Schaider, L. Showe, K. Satyamoorthy, P. Robbins, and M. Herlyn Transforming Growth Factor-{beta}1 Increases Survival of Human Melanoma through Stroma Remodeling Cancer Res., November 1, 2001; 61(22): 8306 - 8316. [Abstract] [Full Text] [PDF]
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O. Straume and L. A. Akslen Expresson of Vascular Endothelial Growth Factor, Its Receptors (FLT-1, KDR) and TSP-1 Related to Microvessel Density and Patient Outcome in Vertical Growth Phase Melanomas Am. J. Pathol., July 1, 2001; 159(1): 223 - 235. [Abstract] [Full Text] [PDF]
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I. Feoktistov, A. E. Goldstein, S. Ryzhov, D. Zeng, L. Belardinelli, T. Voyno-Yasenetskaya, and I. Biaggioni Differential Expression of Adenosine Receptors in Human Endothelial Cells: Role of A2B Receptors in Angiogenic Factor Regulation Circ. Res., March 22, 2002; 90(5): 531 - 538. [Abstract] [Full Text] [PDF]
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