| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Department of Internal Medicine, Division of Pulmonary and Critical Care Medicine, University of Michigan Medical School, Ann Arbor, MI 48109
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Multiple studies have demonstrated the association between increased tumor vascularity and macrophage infiltration (16, 17, 18, 19), suggesting that the presence of macrophages enhances the angiogenic potential of tumors. It also has been shown that tumor samples with higher macrophage counts tend to be associated with poorer patient prognosis (19, 20, 21). These observations suggest that the presence of macrophages within tumors may promote angiogenic activity. Furthermore, it may be the interaction between tumor cells and macrophages that induces macrophages to produce angiogenic factors.
Because macrophages are potentially important sources of angiogenic factors in cancer, we hypothesized that interaction between non-small cell lung cancer (NSCLC)3 and peripheral blood monocytes (PBM) would enhance the angiogenic potential of tumors. In this study, we show that although NSCLC and PBM both induce endothelial cell chemotaxis, coculture of PBM with NSCLC leads to a synergistic increase in angiogenic activity that is attributable to increased expression of angiogenic CXC chemokines. Furthermore, we have found that macrophage migration-inhibitory factor (MIF) is, in large part, the NSCLC-derived factor which accounts for the increased macrophage-derived angiogenic activity.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Human lung non-small cell carcinoma cells (A549; ATCC CCL-185; American Type Culture Collection, Manassas, VA) and human PBM were maintained in RPMI 1640, 10% FCS supplemented with 100 U/ml penicillin, 100 µg/ml streptomycin, 1% L-glutamine, and 2.5% HEPES buffer. A second human lung carcinoma cell line (Calu 6; ATCC HTB-56) was maintained in Eagle’s minimum essential medium/10% FCS with 0.1 mM nonessential amino acids and 1 mM sodium pyruvate. Human dermal microvascular endothelial cells (HMVEC) (Cell Systems, Kirkland, WA) were maintained per the supplier’s recommendations in CS-C complete medium. Human microvascular endothelial cells (HMEC-1) kindly provided by Dr. E. Ades and Mr. F. J. Candal (Center for Disease Control, National Center for Infectious Diseases, Atlanta, GA) and Dr. T. Lawley (Emory University, Atlanta, GA), were maintained in MCDB131 medium (Life Technologies, Grand Island, NY) to which FCS (10%), 100 U/ml penicillin, 100 µg/ml streptomycin, 1% L-glutamine, 10 ng/ml epidermal growth factor, and 1 µg/ml hydrocortisone had been added. Medium was changed in all cell lines every 48–72 h. All cell lines were incubated at 37°C with 5% CO2.
Purified and biotinylated Ab for epithelial-neutrophil activating
peptide (ENA)-78, growth-related oncogene (GRO)-
, IFN-
-inducible
protein (IP)-10, monokine induced by IFN- (MIG), MIF, and vascular
endothelial growth factor (VEGF) were obtained from R&D Systems
(Minneapolis, MN). Abs for GRO- were obtained from PeproTech (Rocky
Hill, NJ). Purified and biotinylated Abs for IL-8 were produced as
described previously (22).
Monocyte isolation
After informed consent was obtained, whole blood was obtained by
venipuncture from healthy donors and anticoagulated with 1000 U
heparin/60 ml blood drawn. Mononuclear cells were isolated by
Ficoll-Paque (Amersham Pharmacia Biotech, Uppsala, Sweden) gradient
centrifugation. Remaining red blood cells were lysed by washing with
ACK buffer (1 mM KHCO3, 0.15 mM
NH4Cl, and 0.1 mM Na2EDTA).
Resultant mononuclear cells (
30% monocytes and 70% lymphocytes)
were washed twice with HBSS without Mg2+ or
Ca2+, and resuspended in serum-free RPMI 1640
with 0.1% BSA at a concentration of 3 x
106 cells/ml. To generate monocyte-conditioned
medium (CM), 500 µl of the suspension was plated out in each well of
a 24-well plate. Plates were incubated at 37°C with 5%
CO2 for 1 h, and nonadherent cells were
removed by washing with RPMI 1640. Wells then were filled with 500 µl
of serum-free RPMI 1640, and specimens were incubated for 48 h at
37°C with 5% CO2. At 48 h, CM was
harvested and centrifuged to remove any cell debris. All samples were
stored at -20°C until used in assays.
Monocyte/NSCLC coculture
A549 or Calu 6 cells were grown to 80–90% confluence in six-well plates. Cells then were washed twice with serum-free medium, and PBM (isolated as above) were added into the upper chamber of a 25-mm transwell insert (Nunc, Naperville, IL) and allowed to adhere for 1 h. Cells were washed twice with RPMI 1640 and then transwell inserts were placed directly into A549-containing wells. The upper chamber then was filled with 1 ml of serum-free medium. Cocultures were incubated for 48 h at 37°C with 5% CO2. At 48 h, CM was harvested and centrifuged at 10,000 x g for 5 min. All CM was stored at -20°C until used for the various assays. In some experiments, PBM were exposed to NSCLC CM, and NSCLC cells were exposed to PBM CM.
Endothelial cell chemotaxis assay
Chemotaxis membrane filters (5.0-µm pore size; Osmonics, Livermore, CA) were first prepared by bathing in 2.8% glacial acetic acid for 24 h, followed by two 1-h rinses in deionized, distilled water. The filters then were transferred to a 0.01% gelatin bath and allowed to soak for 24 h. Filters were dried under a laminar flow hood and stored at room temperature for up to 1 month. Endothelial cell chemotaxis assays were performed in 12-well, blind well chemotaxis chambers (Neuroprobe, Cabin John, MD). Briefly, human endothelial cells were suspended at a concentration of 106 cells/ml in their respective serum-free medium (in the case of HMEC-1, MCDB-131 medium was supplemented with 0.1% BSA). Endothelial cells were placed into each of the bottom wells (165 µl). Membranes were placed over the wells, the gasket applied, and the chambers sealed. Chambers were inverted and incubated at 37°C with 5% CO2 for 1 h (HMEC-1) or 2 h (HMVEC) to allow for endothelial cell adherence to the membrane. Chambers then were reinverted and the CM to be studied was placed in the upper well (116 µl). Chambers were reincubated for 2 h. Membranes then were scraped to remove any adherent, unmigrated endothelial cells from the lower chamber, fixed in methanol, stained with a modified Wright-Giemsa stain, and cells that had migrated through the membrane were counted in five high-power fields (hpf; x200). Results were expressed as the mean number of endothelial cells that had migrated per hpf ± SEM. Each sample was assessed in triplicate. Experiments were performed at least three times. Results obtained were similar whether HMEC-1 or HMVEC cells were used. Each experiment was accompanied by a positive control (VEGF 100 ng/ml) and a negative control (nonconditioned medium) to account for variable chemotaxis from one assay to the next (data not shown).
ELISA
CXC chemokines (IL-8 (CXCL8), ENA-78 (CXCL5), GRO- (CXCL1),
MIG, (CXCL9); and IP-10, (CXCL10)), MIF, and VEGF were quantitated by a
double-ligand method described previously (5). In brief,
flat-bottom 96-well microtiter plates (Immuno Plate F96; Nunc) were
coated with the appropriate purified Abs overnight at 4°C and were
then washed three times with PBS with 0.05% Tween 20 (washing buffer).
Nonspecific binding sites were blocked with PBS and 2% BSA. After
washing three times, samples (neat and 1:10 dilution) and standards
were added as 50-µl aliquots and incubated at 37°C. Plates were
washed three times and incubated with the respective biotinylated Ab at
37°C. Plates were again washed three times and incubated with
polyperoxidase-avidin substrate (Endogen, Woburn, MA) at 37°C.
Subsequently, plates were incubated with the peroxidase substrate
(Dako, Carpinteria, CA) at room temperature to the desired extinction.
The reaction was terminated with 0.5 M
H2SO4. Plates were read at
490 nm in an ELISA reader. Standards were prepared as 1/2 log
dilutions of purified recombinant Ab, from 100 ng/ml to 0.001 ng/ml per
well. For VEGF and MIF, we developed an ELISA by performing multiple
assays on standards of human recombinant protein from 1pg/ml to
100ng/ml using varying concentrations of coating and detecting Abs
until optimal working concentrations were defined. VEGF ELISA used 20
ng/ml for coating and 75 ng/ml for detecting, and consistently and
specifically detected quantities of VEGF121 and
VEGF165 (the predominant secretory forms of human
VEGF) at concentrations as low as 1 pg/ml. For MIF, 75 ng/ml was used
for coating, and 20ng/ml was used for detection. This ELISA had a
similar sensitivity and specificity (data not shown).
Immunodepletion of MIF from NSCLC CM
A total of 250 mg of Protein A immobilized on Sepharose beads (Sigma, St. Louis, MO) was reconstituted in 15 ml of PBS (pH 8.0) overnight at 4°C, washed twice, then brought up in PBS to a final concentration of 150 mg/ml. A total of 1 µg of goat anti-human MIF or goat IgG (R&D Systems) was added to 100 µl of the protein A-Sepharose solution and brought up to a total volume of 500 µl in PBS, mixed thoroughly, and incubated for 2 h at 4°C. The beads then were washed and resuspended in 100 µl of PBS. Then1 ml of A549 or Calu 6 CM was added to the beads and incubated on a rocker plate for 3 h at 4°C. The precipitate then was removed by centrifugation at 7500 x g for 5 min. Supernatants then were removed and used to stimulate adherence purified PBMs. One aliquot of each CM was subjected to specific ELISA to confirm that MIF had been depleted by the immunoprecipitation.
Statistical analyses
All generated data were compared by Student’s t test for unpaired observations, and were considered significant if p values were < 0.05. Results were presented as the mean number of migrated endothelial cells/hpf ± SEM. All data are representative of a minimum of three separate experiments. Data were analyzed on a Dell computer with the StatView 5.0.1 statistical software package (SAS Institute, Cary, NC).
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
We hypothesized that because monocyte-derived macrophages are
found within tumors, these two cell types may interact to increase
angiogenic activity. To test this hypothesis, we performed endothelial
cell chemotaxis assays on medium generated from the coculture of PBMs
and NSCLC cell lines. In comparison to CM from PBM alone, A549 CM
alone, or a combination of the two separately generated CM, coculture
CM demonstrated a marked increase in angiogenic activity as assessed by
endothelial cell chemotaxis (p < 0.002; Fig. 1
A). Similarly, PBM CM alone,
Calu 6 CM alone, or separately generated Calu 6 and PBM CM demonstrated
significantly less angiogenic activity than coculture CM
(p < 0.0005; Fig. 1B).
|
To determine the degree to which CXC chemokines or VEGF were
responsible for the increased endothelial cell chemotaxis seen in NSCLC
and PBM CM, we assessed levels of the angiogenic CXC chemokines IL-8
(CXCL8), ENA-78 (CXCL5), GRO- (CXCL1), and angiostatic CXC
chemokines IP-10 (CXCL10) and MIG (CXCL9) as well as VEGF by ELISA.
Coculture of PBM with NSCLC resulted in a marked increase in angiogenic
CXC chemokine expression with a much smaller change in VEGF levels. In
contrast, expression of angiostatic CXC chemokines remained very low
(Table I). LPS contamination was excluded
as a cause of increased CXC chemokine expression by adding polymyxin
B to coculture, which had no effect on NSCLC-induced CXC
chemokine expression from PBMs. LPS levels were below the limit of
detection by Limulus lysate assay (data not shown).
|
We hypothesized that the increased endothelial cell chemotaxis to
PBM/NSCLC coculture was attributable to the observed increase in
angiogenic CXC chemokines. To test this hypothesis, endothelial cell
chemotaxis was performed with PBM/NSCLC coculture CM in the presence of
neutralizing Ab to IL-8 (CXCL8), ENA-78 (CXCL5), and GRO- (CXCL1).
We found that endothelial cell chemotaxis to A549/PBM coculture CM was
significantly reduced in the presence of neutralizing Abs to either
IL-8 (CXCL8), ENA-78 (CXCL5), or GRO- (CXCL1) as compared with
control IgG (p < 0.0007; Fig. 2A). Similarly, endothelial
cell chemotaxis to Calu 6/PBM coculture CM in the presence of
neutralizing Ab to IL-8 (CXCL8) or GRO- (CXCL1) was significantly
less than in the presence of control IgG (p <
0.002; Fig. 2B). Recently, CXCR2 was identified as the
receptor mediating the angiogenic activity of CXC chemokines
(23). Therefore, we performed endothelial cell chemotaxis
to coculture CM using endothelial cells preincubated with neutralizing
Ab to CXCR2 or control IgG. We found neutralizing Ab to CXCR2 markedly
reduced the increased angiogenic response to coculture CM (Calu 6/PBM
or A549/PBM; Fig. 2C; p < 0.0001 for both
conditions).
|
We demonstrated that angiogenic CXC chemokine levels are markedly
increased when PBM are cocultured with NSCLC. To determine the source
of the increased chemokine levels, we first incubated PBM in cell-free
NSCLC CM and NSCLC in cell-free PBM CM to assess chemokine expression.
We found that when PBM are incubated with CM from NSCLC, there is a
significant increase in expression of the angiogenic CXC chemokines
similar to that seen when PBM are cocultured with NSCLC cells (Table II). Interestingly, NSCLC cells cultured
in cell-free PBM CM also expressed increased levels of angiogenic CXC
chemokines as compared with baseline expression (Tables I
and II),
albeit to a much lesser degree. This suggests that PBM are the
predominant source of the increased chemokine expression in coculture.
All CM were tested and found to be free of endotoxin by
Limulus lysate assay (data not shown).
|
Recently, MIF has been associated with tumor angiogenesis
(24, 25, 26, 27), but it has not been shown to be directly
angiogenic. We hypothesized that MIF may be the tumor-derived factor
that induces angiogenic activity from PBMs. To test this hypothesis, we
first measured levels of MIF in 48-h CM from unstimulated NSCLC cell
lines. MIF was detected in both A549 (4.8 ± 0.2 ng/ml) and Calu 6
CM (6.9 ± 1.0 ng/ml). To determine whether MIF was responsible
for increased PBM-derived angiogenic activity, we performed endothelial
cell chemotaxis assays on CM from PBMs that had been exposed to Calu 6
or A549 CM. Before stimulation of PBMs, NSCLC CM was subjected to
immunodepletion with either IgG or goat anti-human MIF Ab.
Depletion of MIF was confirmed by ELISA (data not shown). We found that
NSCLC CM depleted of MIF was significantly less effective at inducing
PBM-derived angiogenic activity (p < 0.001;
Fig. 3). Furthermore, PBM stimulated with
MIF-depleted A549 or Calu 6 CM resulted in significantly decreased CXC
chemokine expression compared with control A549 or Calu 6 CM
(p < 0.05 for each condition; Fig. 4).
|
|
The above data suggests that MIF is necessary for NSCLC cell
line-induction of CXC chemokine-dependent angiogenic activity from
PBMs. To determine whether MIF was sufficient to induce CXC chemokines,
freshly isolated PBM were cultured in six-well plates in the presence
of serum-free medium with or without the addition of 10 ng/ml MIF. CM
was harvested at 24 h and assayed for IL-8, ENA-78, and GRO- by
specific ELISA. We found that PBM stimulated with MIF expressed
significantly higher levels of IL-8 (252.7 ± 16.5 ng/ml vs
54.6 ± 2.0 ng/ml; p < 0.01), ENA-78 (264.8
± 26.9 ng/ml vs 105.2 ± 6.7 ng/ml; p < 0.01),
and GRO- (7.9 ± 1.2 ng/ml vs 1.9 ± 0.4 ng/ml;
p < 0.01) compared with unstimulated PBM (Fig. 5).
|
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
A number of different cell types contribute to angiogenesis, including cancer cells (12, 34), endothelial cells (35, 36, 37), fibroblasts (13, 15), eosinophils (38), platelets (39), and monocyte-derived macrophages (40, 41, 42). Each cell type displays a certain repertoire of angiogenic factors, depending on the stimulus. Because of the overwhelming redundancy within this system, it is important to determine the cellular source(s) of angiogenic activity in pathologic conditions. In this study, we found that PBM are induced by NSCLC cells to increase expression of angiogenic activity. In addition, we observed some increase in expression of angiogenic factors by NSCLC cells exposed to PBM CM.
Many authors have cited similarities between tumor growth and wound
healing (43, 44, 45). Macrophages are critically involved in
wound repair (46, 47, 48). In wounds, local tissue injury
results in leakage of plasma and blood from damaged blood vessels
resulting in a fibrin clot rich in fibronectin, fibrin breakdown
products, and platelets. The fibrin clot becomes a provisional matrix
or stroma into which monocytes migrate to ingest wound debris
(43). Monocyte-derived macrophages are essential in this
role, as evidenced by defective wound repair in macrophage-depleted
animals (49). In addition to their role in removing
debris, macrophages in inflammatory conditions promote tissue repair by
producing cytokines such as TNF- (50), platelet-derived
growth factor (51), and VEGF (52). Analogous
events are thought to occur in the process of tumor angiogenesis
(43, 44, 45, 46).
Polverini and Leibovich (52) demonstrated that
tumor-associated macrophages or their CM isolated from experimental
fibrosarcoma display angiogenic activity, suggesting the role of
macrophage-derived soluble factors in tumor angiogenesis. Similarly,
Hildenbrand et al. (53) showed that TGF-
induced
macrophages to produce urokinase-plasminogen activator, a known factor
in the degradation of tumor matrix and tumor progression.
Interestingly, this group also demonstrated that tumor-associated
macrophages were induced to produce significantly more
urokinase-plasminogen activator than either PBM or elicited tissue
macrophages (53), suggesting that the tumor environment
can alter macrophage phenotype.
We recently have demonstrated infiltration of monocytes in human NSCLC, which is mediated by tumor expression of CC chemokines (20). Because PBM are a potentially rich source of angiogenic factors, including angiogenic CXC chemokines (41, 55, 56), we hypothesized that NSCLC may augment the expression of macrophage-derived angiogenic factors. Our results show that NSCLC cells promote angiogenesis by inducing host PBM to up-regulate CXC chemokine-dependent angiogenic activity. This is important in the context of our previous findings demonstrating the role of the CXC chemokine family in NSCLC-mediated angiogenesis (5, 10, 31). Although it is likely that other factors such as VEGF play an important role in NSCLC angiogenesis, the results of our study do not support a predominant role for VEGF in the monocyte-derived angiogenic activity induced by NSCLC.
Recently, Addison et al. have demonstrated that CXCR2 is the putative receptor for the angiogenic activity mediated by angiogenic CXC chemokines (23). In our study, we found that the increased angiogenic activity in coculture CM could be neutralized by Abs directed against CXCR2. It is interesting to note that the degree of inhibition of angiogenic activity by anti-CXCR2 was comparable to, or only slightly greater than, the inhibition seen with Abs directed at the individual ligands. To account for this observation, one can speculate that Ab binding to an individual CXC chemokine ligand may prevent access to the receptor by other ligands, thereby effectively neutralizing the receptor as well.
We found that MIF, expressed constitutively by the A549 and Calu 6 cell lines, is responsible in large part for the induction of PBM-derived angiogenic activity. MIF was initially discovered as an activity released by cells in delayed-type hypersensitivity reactions. Recently, a variety of immunomodulatory actions have been attributed to MIF (56). Interestingly, several groups of investigators have found that MIF can promote angiogenic activity in mouse models of tumor growth (25, 26). However, data demonstrating direct angiogenic activity for MIF is lacking. These reports prompted us to investigate whether NSCLC cells expressed MIF and whether this might be a mechanism by which NSCLC cells induce PBM-derived angiogenic activity. We found constitutive expression of MIF protein in the CM of A549 and Calu 6 cells. Furthermore, we demonstrated that MIF was responsible in part for the induction of PBM-derived CXC chemokine-dependent angiogenic activity by NSCLC cells.
We observed that the degree of PBM stimulation by NSCLC cell lines was greater when CM was used as the stimulus compared with coculture conditions. It is interesting to speculate on the existence of a simultaneous inhibitory signal generated by NSCLC cells and present in soluble form. This inhibitory signal must be postulated to have a short biologic half-life in vitro, given that its effect is only seen in coculture conditions. When CM is generated separately and used to stimulate PBMs, a mediator of short half-life would have less of an effect in the absence of continued secretion.
Combined with our previous findings of CC chemokine-mediated monocyte infiltration of NSCLC tumors (20), our current data suggest that tumors orchestrate a complex cytokine network in which macrophages recruited to the tumor site are influenced to promote a proangiogenic milieu. Our findings may account for the antitumor activity of neutralizing Abs to MIF in animal models of cancer as reported by other investigators (25, 26, 27).
|
Footnotes |
|---|
2 Address correspondence and reprint requests to Dr. Douglas Arenberg, University of Michigan Medical Center, Division of Pulmonary and Critical Care Medicine, 6301 MSRB III, Box 0642, 1150 West Medical Center Drive, Ann Arbor, MI 48109. E-mail address: darenber{at}umich.edu
3 Abbreviations used in this paper: NSCLC, non-small cell lung cancer; PBM, peripheral blood monocyte; MIF, migration-inhibitory factor; HMVEC, human dermal microvascular endothelial cells; HMEC, human microvascular endothelial cells; CM, conditioned medium; ENA, epithelial neutrophil-activating peptide; GRO, growth-related oncogene; IP, IFN--inducible protein; MIG, monokine induced by IFN-; VEGF, vascular endothelial growth factor; hpf, high-power field.
Received for publication November 16, 2000. Accepted for publication April 13, 2001.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
-inducible protein 10 (IP-10) is an angiostatic factor that inhibits human non-small cell lung cancer (NSCLC) tumorigenesis and spontaneous metastases. J. Exp. Med. 184:981. as a critical macrophage chemoattractant in murine wound repair. J. Clin. Invest. 101:1693.[Medline]
stimulates urokinase expression in tumor-associated macrophages of the breast. Lab. Invest. 78:59.[Medline]
This article has been cited by other articles:
|
|
 |
|
  J. Martin, F. J. Duncan, T. Keiser, S. Shin, D. F. Kusewitt, T. Oberyszyn, A. R. Satoskar, and A. M. VanBuskirk Macrophage migration inhibitory factor (MIF) plays a critical role in pathogenesis of ultraviolet-B (UVB) -induced nonmelanoma skin cancer (NMSC) FASEB J, March 1, 2009; 23(3): 720 - 730. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. McClelland, L. Zhao, S. Carskadon, and D. Arenberg Expression of CD74, the Receptor for Macrophage Migration Inhibitory Factor, in Non-Small Cell Lung Cancer Am. J. Pathol., February 1, 2009; 174(2): 638 - 646. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Frascaroli, S. Varani, N. Blankenhorn, R. Pretsch, M. Bacher, L. Leng, R. Bucala, M. P. Landini, and T. Mertens Human Cytomegalovirus Paralyzes Macrophage Motility through Down-Regulation of Chemokine Receptors, Reorganization of the Cytoskeleton, and Release of Macrophage Migration Inhibitory Factor J. Immunol., January 1, 2009; 182(1): 477 - 488. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-Y. Lee, H.-F. Sher, H.-W. Chen, C.-C. Liu, C.-H. Chen, C.-S. Lin, P.-C. Yang, H.-S. Tsay, and J. J.W. Chen Anticancer effects of tanshinone I in human non-small cell lung cancer Mol. Cancer Ther., November 1, 2008; 7(11): 3527 - 3538. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Coleman, B. E. Rendon, M. Zhao, M.-W. Qian, R. Bucala, D. Xin, and R. A. Mitchell Cooperative Regulation of Non-Small Cell Lung Carcinoma Angiogenic Potential by Macrophage Migration Inhibitory Factor and Its Homolog, D-Dopachrome Tautomerase J. Immunol., August 15, 2008; 181(4): 2330 - 2337. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Zhou, X. Yan, J. Gershan, R. J. Orentas, and B. D. Johnson Expression of Macrophage Migration Inhibitory Factor by Neuroblastoma Leads to the Inhibition of Antitumor T Cell Reactivity In Vivo J. Immunol., August 1, 2008; 181(3): 1877 - 1886. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. E. Rendon, T. Roger, I. Teneng, M. Zhao, Y. Al-Abed, T. Calandra, and R. A. Mitchell Regulation of Human Lung Adenocarcinoma Cell Migration and Invasion by Macrophage Migration Inhibitory Factor J. Biol. Chem., October 12, 2007; 282(41): 29910 - 29918. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. R. McClelland, S. L. Carskadon, L. Zhao, E. S. White, D. G. Beer, M. B. Orringer, A. Pickens, A. C. Chang, and D. A. Arenberg Diversity of the Angiogenic Phenotype in Non-Small Cell Lung Cancer Am. J. Respir. Cell Mol. Biol., March 1, 2007; 36(3): 343 - 350. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Wald, U. Izhar, G. Amir, S. Avniel, Y. Bar-Shavit, H. Wald, I. D. Weiss, E. Galun, and A. Peled CD4+CXCR4highCD69+ T Cells Accumulate in Lung Adenocarcinoma J. Immunol., November 15, 2006; 177(10): 6983 - 6990. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. D. Swant, B. E. Rendon, M. Symons, and R. A. Mitchell Rho GTPase-dependent Signaling Is Required for Macrophage Migration Inhibitory Factor-mediated Expression of Cyclin D1 J. Biol. Chem., June 17, 2005; 280(24): 23066 - 23072. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.-L. Yao, Y.-C. Lin, C.-H. Wang, Y.-C. Huang, W.-Y. Liao, S.-S. Wang, J. J. W. Chen, and P.-C. Yang Autocrine and Paracrine Regulation of Interleukin-8 Expression in Lung Cancer Cells Am. J. Respir. Cell Mol. Biol., June 1, 2005; 32(6): 540 - 547. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J.W. Chen, Y.-C. Lin, P.-L. Yao, A. Yuan, H.-Y. Chen, C.-T. Shun, M.-F. Tsai, C.-H. Chen, and P.-C. Yang Tumor-Associated Macrophages: The Double-Edged Sword in Cancer Progression J. Clin. Oncol., February 10, 2005; 23(5): 953 - 964. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. M. Langenfeld and J. Langenfeld Bone Morphogenetic Protein-2 Stimulates Angiogenesis in Developing Tumors Mol. Cancer Res., March 1, 2004; 2(3): 141 - 149. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Campa, S. Zienolddiny, V. Maggini, V. Skaug, A. Haugen, and F. Canzian Association of a common polymorphism in the cyclooxygenase 2 gene with risk of non-small cell lung cancer Carcinogenesis, February 1, 2004; 25(2): 229 - 235. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Amin, O. V. Volpert, J. M. Woods, P. Kumar, L. A. Harlow, and A. E. Koch Migration Inhibitory Factor Mediates Angiogenesis via Mitogen-Activated Protein Kinase and Phosphatidylinositol Kinase Circ. Res., August 22, 2003; 93(4): 321 - 329. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J.W. Chen, P.-L. Yao, A. Yuan, T.-M. Hong, C.-T. Shun, M.-L. Kuo, Y.-C. Lee, and P.-C. Yang Up-Regulation of Tumor Interleukin-8 Expression by Infiltrating Macrophages: Its Correlation with Tumor Angiogenesis and Patient Survival in Non-Small Cell Lung Cancer Clin. Cancer Res., February 1, 2003; 9(2): 729 - 737. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. S. White, K. R. Flaherty, S. Carskadon, A. Brant, M. D. Iannettoni, J. Yee, M. B. Orringer, and D. A. Arenberg Macrophage Migration Inhibitory Factor and CXC Chemokine Expression in Non-Small Cell Lung Cancer: Role in Angiogenesis and Prognosis Clin. Cancer Res., February 1, 2003; 9(2): 853 - 860. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Tomiyasu, I. Yoshino, R. Suemitsu, T. Okamoto, and K. Sugimachi Quantification of Macrophage Migration Inhibitory Factor mRNA Expression in Non-Small Cell Lung Cancer Tissues and Its Clinical Significance Clin. Cancer Res., December 1, 2002; 8(12): 3755 - 3760. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
A. Barchowsky, N. V. Soucy, K. A. O'Hara, J. Hwa, T. L. Noreault, and A. S. Andrew A Novel Pathway for Nickel-induced Interleukin-8 Expression J. Biol. Chem., June 28, 2002; 277(27): 24225 - 24231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. S. White, D. L. Livant, S. Markwart, and D. A. Arenberg Monocyte-Fibronectin Interactions, Via {alpha}5{beta}1 Integrin, Induce Expression of CXC Chemokine-Dependent Angiogenic Activity J. Immunol., November 1, 2001; 167(9): 5362 - 5366. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
