(Hypertension. 2001;37:581.)
© 2001 American Heart Association, Inc.
Scientific Contributions |
Mitogenic and Antiapoptotic Actions of Hepatocyte Growth Factor Through ERK, STAT3, and Akt in Endothelial Cells
From the Department of Geriatric Medicine (H.N., R.M., K.Y., Y.T., M.A., T.O.), the Division of Gene Therapy Science (R.M., Y.K.), and the Division of Biochemistry, Department of Oncology, Biomedical Research Center (K.M., T.N.), Graduate School of Medicine, Osaka University, Osaka, Japan, and the Department of Medical Biochemistry (H.N., M.H.), Ehime University School of Medicine, Ehime, Japan.
Correspondence to Ryuichi Morishita, MD, PhD, Department of Geriatric Medicine, Osaka University Medical School, 2 -2 Yamada-oka, Suita 565 to 0871, Japan. E-mail morishit{at}geriat.med.osaka-u.ac.jp
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Abstract |
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Hepatocyte growth factor (HGF), a member of the angiogenic growth factors, may play a pivotal role in the regulation of endothelial cells, inasmuch as HGF shows mitogenic and antiapoptotic actions in endothelial cells. Because the mechanism of these actions is still unclear, we examined the signal transduction system of HGF in human aortic endothelial cells. Treatment of endothelial cells with recombinant HGF (rHGF) resulted in a significant increase in DNA synthesis as assessed by thymidine incorporation. Importantly, phosphorylation of extracellular signal–related kinase (ERK) and Akt by rHGF was clearly observed. Thus, we further examined the effects of specific inhibitors of ERK or Akt on cell proliferation. Pretreatment with PD98059, a mitogen-activated protein kinase kinase inhibitor, significantly attenuated cell proliferation induced by rHGF, whereas inhibitors of phosphatidylinositol-3-OH kinase, wortmannin, and LY-294002, did not. Interestingly, treatment with rHGF significantly increased the phosphorylation of the signal transducers and activators of transcription (STAT)3 (Ser727), whereas PD98059 attenuated the phosphorylation of Ser727 induced by rHGF. In addition, treatment with rHGF significantly increased the promoter activity of c-fos, which includes the sis-inducible element and serum response element, whereas PD98059 completely attenuated the activation of the c-fos promoter induced by rHGF. In contrast, inhibition of Akt by wortmannin and LY-294002 failed to inhibit the phosphorylation of STAT3 and c-fos activation. On the other hand, treatment with rHGF attenuated the increase in LDH release and caspase-3 activity induced by tumor necrosis factor-
stimulation. In
contrast to DNA synthesis, wortmannin and LY-294002 markedly attenuated
the decrease in caspase-3 activity mediated by rHGF, whereas PD98059
did not. Overall, the present study demonstrated that HGF
stimulated cell proliferation through the ERK-STAT3 (Ser727) pathway
and had an antiapoptotic action through the
phosphatidylinositol-3-OH kinase–Akt pathway in human aortic
endothelial cells. These findings provide new
perspectives in the role of HGF in cardiovascular
disease.
Key Words: vascular • growth substances • kinase • apoptosis • transcription
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Introduction |
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Dysfunction of endothelial cells may promote abnormal vascular growth, such as that found in atherosclerosis and arteriosclerosis, including hypertensive complications. From this viewpoint, we have focused on the role of angiogenic growth factors, such as vascular endothelial growth factor and basic fibroblast growth factor in endothelial cells. In addition, we and others have identified that hepatocyte growth factor (HGF) is also a member of angiogenic growth factors.1 2 3 4 These angiogenic growth factors induce the proliferation and migration of endothelial cells, remodeling of the extracellular matrix, and the formation of capillary tubules.5
HGF is a multifunctional cytokine possessing a wide spectrum of biological activities. It is secreted by cells of mesenchymal origin and acts as a mitogen, dissociation factor, and motility factor for many epithelial cells in culture6 7 8 through its tyrosine kinase receptor, c-met.9 We reported that HGF linking to c-met acts as a protective factor against endothelial cell death induced by serum-free treatment or high glucose conditions.10 11 12 Various intracellular signaling pathways have been shown to be activated by tyrosine kinases linked to c-met.13 The biological responses mediated by c-met are triggered by the tyrosine phosphorylation of a single multifunctional docking site located in the carboxy-terminal tail of the receptor.14 This sequence, containing 2 phosphotyrosines, interacts with several cytoplasmic signal transducers either directly or indirectly through molecular adapters such as Grb2, Shc, and Gab1.15 16 It has been reported that after HGF stimulation, c-met binds and activates phosphatidylinositol-3-OH (PI3) kinase and recruits the Grb-SOS complex, stimulating the Ras–mitogen-activated protein (MAP) kinase cascade.17 18 19 Although HGF has an antiapoptotic action under serum-free conditions through extracellular signal–related kinase (ERK) activation,12 the critical signal pathways for cell proliferation and the antiapoptotic effect of HGF are still largely unknown. Therefore, we focused on the signal transduction pathway of HGF in human aortic endothelial cells.
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Methods |
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Cell Culture
Human aortic endothelial cells (passage 3) were obtained from Clonetics Corp and cultured in modified MCDB131 medium supplemented with 5% FCS, 50 µg/mL gentamicin sulfate, 50 ng/mL amphotericin B, 10 ng/mL epidermal growth factor, and 1 mmol/L hydrocortisone in the standard fashion.20 Cells were incubated at 37°C in a humidified atmosphere of 95% air/5% CO2 with medium changes every 2 days. These cells showed the specific characteristics of endothelial cells by immunohistochemical examination and morphological observation. Briefly, human aortic endothelial cells tested positively for factor VIII antigen and for uptake of diacetylated LDL. All the cells were used within passages 5 to 8.
Measurement of DNA Synthesis as Assessed by
Thymidine Incorporation
Endothelial cells in growth medium
were equally seeded into 24-well culture plates. The next day, the
growth medium was changed to medium supplemented with 0.5% FCS. On day
3 after seeding, 1 hour before the addition of HGF,
inhibitors were added to each well of the culture plate,
and then the medium was changed to fresh serum-free medium containing
HGF (100 ng/mL) or vehicle, and the plate was incubated for 36 hours.
Twelve hours before harvest, [3H]thymidine
(1 µCi/µL, Amersham Pharmacia Biotech UK) was added to each well of
the culture plate. Before the count, the DNA was precipitated with cold
10% trichloroacetic acid for 20 minutes, and the precipitated material
was resuspended in 0.2 mL of 0.3 mol/L NaOH. The cells were harvested,
thoroughly washed, and assayed for
[3H] in a liquid scintillation
counter.
Western Blotting
Western blotting was performed for analysis
of ERK, Akt, and signal transducers and activators of
transcription (STAT)3 by using a phosphospecific antibody as previously
described.12 After treatment,
the cells were extracted with lysis buffer (50 mmol/L Tris-Cl,
2.5 mmol/L EGTA, 1 mmol/L EDTA, 10 mmol/L NaF, 1%
deoxycorticosterone, 1% Triton X-100, 1 mmol/L
phenylmethylsulfonyl fluoride, and 2 mmol/L
Na3VO4). Samples
containing 20 µg protein were run on 10% SDS-polyacrylamide
gels, separated by SDS-PAGE, transferred to nitrocellulose membranes
(Hybond ECL, Amersham), and incubated with a polyclonal antibody to
phosphospecific or total ERK (anti-human, -rat, -mouse, or -rabbit IgG,
1:1000, Cell Signaling Technology), phosphospecific or total Akt
(anti-human, -rat, -mouse, or -rabbit IgG, 1:1000, Cell Signaling
Technology), or phosphospecific (Tyr705 or Ser727) or total STAT3
(anti-human, -rat, -mouse, or -rabbit IgG, 1:2000, Cell Signaling
Technology) at 4°C overnight. The membranes were then washed and
incubated with a 1:2000 dilution of rabbit immunoglobulin horseradish
peroxidase–conjugated antibody (Amersham). Bound antibodies were
detected by enhanced chemiluminescence (ECL, Amersham) and
Hyperfilm-MP (Amersham). To
quantify and compare levels of proteins, the density of each band was
measured by densitometry (Shimazu).
c-fos
Promoter Assay
Endothelial cells were seeded in
6-well plates and transfected with
c-fos–luciferase reporter gene
(p2FTL) by using lipofectAMINE PLUS (GIBCO-BRL) as previously
described.21 The
fos-luciferase reporter gene consists of 2 copies of the
c-fos 5'-regulated enhancer
element (-357 to -276), the herpes simplex virus thymidine kinase
gene promoter (-200 to +70), and the luciferase
gene.22 At 24 hours after
transfection, transfected cells were incubated with serum-free medium
for 24 hours. Quiescent cells were treated with 100 ng/mL HGF for 4
hours, washed with PBS, and lysed for 15 minutes with 500 µL cell
lysis buffer at room temperature. Then, 10 µL cell extract was mixed
with 100 µL luciferase assay reagent, and the light produced was
measured for 30 seconds with use of a
luminometer.
LDH Release
The extent of cell death was assessed by using a kit
(Wako) to measure released LDH activity from dead cells, because loss
of cell membrane integrity was observed in both necrotic and
apoptotic cells.23
After subconfluence was attained, the medium was changed to serum-free
medium. The next day, the medium was changed to fresh serum-free medium
containing HGF or vehicle.
Activity of Caspase-3 Protease
Cells were harvested after exposure to HGF (100
ng/mL) or vehicle for the indicated periods of time, washed 3 times
with PBS, and then suspended in buffer containing 50 nmol/L Tris-HCl
(pH 7.4), 1 mmol/L EDTA, and 10 mmol/L EGTA. After addition
of 10 µmol/L digitonin, cells were incubated at 37°C for 10
minutes. Lysates were centrifuged at
900g for 3 minutes, and the
resulting supernatants (40 µg protein) were incubated with 50
µmol/L enzyme substrate Ac-DEVD-MCA at 37°C for 1 hour. The
level of released 7-amino-4-methylcoumarin was measured
by use of a spectrofluorometer (Hitachi F-3000 or F-2000) with
excitation at 380 nm and emission at 460 nm. Excitation and emission
slit widths were adjusted to 10 and 20 mm,
respectively.
Materials
Human recombinant HGF was purified from the culture
medium of Chinese hamster ovary cells or C-127 cells, which were
transfected with an expression plasmid containing human HGF
cDNA.7 Human recombinant tumor
necrosis factor (TNF)- was obtained from Peprotech. PD98059, a
specific inhibitor of MAP kinase kinase (MEK), was obtained
from New England BioLabs; wortmannin and trichloroacetic acid were
obtained from Sigma Chemical Co; and LY-294002 was obtained from
Calbiochem.
Statistical Analysis
All values are expressed as mean±SEM. ANOVA with a
subsequent Bonferroni/Dunnett test was used to determine the
significance of differences in multiple comparisons. Values of
P<0.05 were considered
statistically significant.
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Results |
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Effect of HGF on ERK (Thr202/Tyr204) and Akt (Ser473) Phosphorylation
The ERK pathway, which is activated by growth factors, is involved in mediating cellular proliferation, transformation, and differentiation. Consistent with previous reports,12 ERK was phosphorylated from 5 minutes after the addition of HGF, and maximal tyrosine phosphorylation was detected at 15 minutes (Figure 1A, P<0.01), whereas total ERK proteins were not altered by treatment with HGF. In contrast, the protein kinase Akt serves as a multifunctional regulator of cell survival, growth, and glucose metabolism.24 With respect to its cardiovascular function, Akt acts downstream of vascular endothelial growth factor25 to confer endothelial cell survival and ensure proper blood vessel development.26 Nevertheless, there is no report about Akt phosphorylation induced by recombinant HGF (rHGF). As shown in Figure 1B, Akt was also phosphorylated 5 minutes after the addition of rHGF (P<0.01).
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Role of Phosphorylation of ERK
or Akt in Mitogenic Activity of HGF
Therefore, in the present study, we further
examined how ERK and Akt act in the mitogenic action of
HGF. In previous reports, treatment with PD98059, a MEK
inhibitor, significantly attenuated
endothelial cell growth induced by rHGF in a
dose-dependent
manner.4 12
Consistent with the previous reports, treatment with PD98059
(30 µmol/L) significantly attenuated DNA synthesis induced by rHGF as
assessed by thymidine incorporation
(Figure 2, P<0.01),
whereas rHGF (100 ng/mL) significantly increased thymidine
incorporation. In contrast, we also examined the effect of 2
structurally unrelated PI3 kinase inhibitors, wortmannin
and LY-294002, on cell proliferation induced by HGF. Wortmannin is a
fungal metabolite that has been characterized as a specific
inhibitor of PI3 kinase at nanomolar
concentrations.27 28
LY-294002 is another specific inhibitor of PI3 kinase at
low micromolar concentrations, but it has no inhibitory
effect against a number of intracellular serine/threonine or tyrosine
kinases at a concentration of 50
µmol/L.29 30 As
shown in
Figure 2, pretreatment with both wortmannin (100 nmol/L) and
LY-294002 (50 µmol/L) could not attenuate the increase in DNA
synthesis induced by rHGF as assessed by thymidine incorporation. These
results demonstrated that HGF stimulated endothelial
growth through ERK rather than Akt
phosphorylation.
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In addition, STAT activity has been reported to be regulated predominantly by phosphorylation on specific tyrosine residues, which causes the STATs to dimerize. STAT dimerization is usually followed by translocation into the nucleus.31 Within the nucleus, STATs recognize and bind to consensus DNA binding sites that represent enhancer sequences for a variety of genes, including immediate early growth response genes, such as c-fos. Because previous studies have demonstrated that HGF increases STAT3 activity in hepatocytes and epithelial cells,32 we examined the phosphorylation of STAT3 (Thy705 and Ser727) induced by rHGF in human endothelial cells. As shown in Figure 3A, treatment with rHGF significantly increased the phosphorylation of STAT3 (Ser727), whereas total STAT3 proteins were not altered by treatment with rHGF. However, unexpectedly, rHGF did not affect the phosphorylation of STAT3 (Thy705). Of importance, pretreatment with PD98059, but not wortmannin or LY-294002, significantly attenuated the phosphorylation of STAT3 (Ser727) induced by rHGF (Figure 3B). Phosphorylation of STAT3 by HGF was further confirmed by the observation that HGF increased luciferase activity driven by sis-inducible element and serum response element (Figure 4), inasmuch as the proto-oncogene c-fos is well known to be induced by STAT3. Of importance, pretreatment with PD98059, but not wortmannin or LY-294002, also significantly attenuated the increase in luciferase activity induced by rHGF (Figure 4, P<0.01).
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Role of Phosphorylation of ERK
or Akt in Antiapoptotic Action of HGF
Because a signaling pathway from PI3 kinase to Akt is
implicated in some cellular responses of PI3 kinase, including
protection from
apoptosis,33 34
we next examined the role of ERK and Akt in the antiapoptotic
action of HGF. Our previous report documented that HGF inhibited cell
death under serum-free conditions, as assessed by LDH
release.12 In the present
study, we used TNF- stimulation as another cell death condition.
Similar to the serum-free condition, the addition of TNF-
significantly increased LDH release
(Figure 5A, P<0.01).
TNF- stimulation also significantly increased caspase-3 activity as
a specific index of apoptosis in a time-dependent manner
(Figure 5B, P<0.01).
Interestingly, treatment with rHGF significantly attenuated the
caspase-3 activation induced by TNF- stimulation
(Figure 5B, P<0.01).
Of importance, pretreatment with PI3 kinase inhibitors,
wortmannin and LY-294002, but not PD98059 significantly attenuated the
decrease in caspase-3 activity induced by rHGF
(Figure 5C, P<0.01).
These results indicate that the Akt pathway is critical in the
antiapoptotic action of HGF as assessed by caspase-3
activity.
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Discussion |
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Vascular structure has been postulated to be determined in large part by the balance between cell growth and cell death by apoptosis. This process of vascular remodeling plays an important role in determining the natural history of vascular disease.35 It has become increasingly clear that the process of cell death by apoptosis is a relatively ubiquitous phenomenon observed in a variety of cell types, including endothelial cells. Because HGF, an angiogenic growth factor, may play a pivotal role in the regulation of endothelial cell growth, we explored the signal transduction system of HGF in endothelial cells. In the present study, we demonstrated that HGF significantly increased DNA synthesis and cell proliferation through the ERK pathway, inasmuch as a MEK inhibitor, PD98059, inhibited the increase in DNA synthesis induced by HGF. Unexpectedly, the Akt pathway seems not to contribute to DNA synthesis induced by HGF.
Recently, a new class of SH2 domain–containing signaling molecules has been found to be involved in mediating the growth-promoting activity of other growth factors, such as platelet-derived growth factor and epidermal growth factor.36 These signaling molecules belong to a family of latent cytoplasmic transcription factors known as STAT. Interestingly, the induction of epithelial tubules by HGF was dependent on activation of the STAT pathway.32 Moreover, c-met, the HGF tyrosine receptor, can bind and directly phosphorylate STAT3.16 32 As discussed earlier, STAT proteins are transcription factors that bind specific DNA elements with high affinity for a modified form of the sis-inducible element, which is part of the c-fos gene promoter. Importantly, the present study demonstrated that HGF phosphorylated STAT3 (Ser727) and activated the c-fos promoter in human endothelial cells, whereas a MEK inhibitor inhibited STAT3 (Ser727) phosphorylation and completely blocked activation of the c-fos promoter induced by rHGF. In contrast, PI3 kinase inhibitors did not inhibit this. From these results, ERK may regulate cell growth at the transcription level of an early response gene, such as c-fos, mediated by STAT3 (Ser727) activation in endothelial cells. Unexpectedly, STAT3 (Thy705) was not phosphorylated by the addition of rHGF in human endothelial cells, but it was phosphorylated in epithelial cells. Phosphorylation of STAT at different sites might affect the physiological response in various cells after HGF stimulation.
In contrast, in the present study,
inhibitors of PI3 kinase could attenuate the
antiapoptotic effect of HGF under TNF- stimulation. Thus,
the present study revealed that HGF exerted an
antiapoptotic action through the PI3 kinase–Akt pathway rather
than the ERK-STAT3 pathway. We speculate that the ERK pathway might be
necessary to maintain endothelial cells and that the
Akt pathway might be more critical in protection against stress injury.
Recent studies have shown that Akt downstream from angiogenic growth
factors functions to promote endothelial cell
survival,24 37 NO
synthesis,38
migration,39 and cellular
responses, which contribute to new blood vessel growth and
stabilization of the vascular network. Akt downstream from the signal
transduction system of HGF is probably involved in the potent
angiogenic activity and antiapoptotic action of HGF.
Nevertheless, the Akt pathway might not be involved in
endothelial growth, because the inhibitors
of PI3 kinase did not affect phosphorylation of STAT3
or activation of the c-fos
promoter. HGF may use different signal molecules in its
mitogenic activity and antiapoptotic action in
human endothelial cells.
Overall, the present study demonstrated that HGF stimulated cell proliferation through the ERK-STAT3 pathway and had an antiapoptotic action through the PI3 kinase–Akt pathway in human aortic endothelial cells. These findings provide new perspectives in the role of HGF in cardiovascular disease, including hypertension, and may explain the preeminent role of HGF in angiogenesis.
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Acknowledgments |
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This work was partially supported by a grant from the Japan Cardiovascular Research Foundation, a Japan Heart Foundation Research Grant, a grant-in-aid from the Tokyo Biochemical Research Foundation, and a grant-in-aid from the Ministry of Education, Science, Sports, and Culture of Japan. We wish to thank Michiko Tamakoshi, Rie Kousai, and Keiko Yamaguchi for their excellent technical assistance.
Received October 24, 2000; first decision November 27, 2000; accepted December 14, 2000.
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  M. L. Guzman-Hernandez, A. Vazquez-Macias, J. Carretero-Ortega, R. Hernandez-Garcia, A. Garcia-Regalado, I. Hernandez-Negrete, G. Reyes-Cruz, J. S. Gutkind, and J. Vazquez-Prado Differential Inhibitor of G{beta}{gamma} Signaling to AKT and ERK Derived from Phosducin-like Protein: EFFECT ON SPHINGOSINE 1-PHOSPHATE-INDUCED ENDOTHELIAL CELL MIGRATION AND IN VITRO ANGIOGENESIS J. Biol. Chem., July 3, 2009; 284(27): 18334 - 18346. [Abstract] [Full Text] [PDF]
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M. Kajiya, H. Shiba, T. Fujita, K. Ouhara, K. Takeda, N. Mizuno, H. Kawaguchi, M. Kitagawa, T. Takata, K. Tsuji, et al. Brain-derived Neurotrophic Factor Stimulates Bone/Cementum-related Protein Gene Expression in Cementoblasts J. Biol. Chem., June 6, 2008; 283(23): 16259 - 16267. [Abstract] [Full Text] [PDF]
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 M. Kanayama, T. Takahara, Y. Yata, F. Xue, E. Shinno, K. Nonome, H. Kudo, K. Kawai, T. Kudo, Y. Tabuchi, et al. Hepatocyte growth factor promotes colonic epithelial regeneration via Akt signaling Am J Physiol Gastrointest Liver Physiol, July 1, 2007; 293(1): G230 - G239. [Abstract] [Full Text] [PDF]
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J. Hur, C.-H. Yoon, C.-S. Lee, T.-Y. Kim, I.-y. Oh, K.-W. Park, J.-H. Kim, H.-S. Lee, H.-J. Kang, I.-H. Chae, et al. Akt Is a Key Modulator of Endothelial Progenitor Cell Trafficking in Ischemic Muscle Stem Cells, July 1, 2007; 25(7): 1769 - 1778. [Abstract] [Full Text] [PDF]
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 A. Bhattacharjee, B. Xu, D. A. Frank, G. M. Feldman, and M. K. Cathcart Monocyte 15-Lipoxygenase Expression Is Regulated by a Novel Cytosolic Signaling Complex with Protein Kinase C {delta} and Tyrosine-Phosphorylated Stat3 J. Immunol., September 15, 2006; 177(6): 3771 - 3781. [Abstract] [Full Text] [PDF]
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H. Nakagami, K. Maeda, R. Morishita, S. Iguchi, T. Nishikawa, Y. Takami, Y. Kikuchi, Y. Saito, K. Tamai, T. Ogihara, et al. Novel Autologous Cell Therapy in Ischemic Limb Disease Through Growth Factor Secretion by Cultured Adipose Tissue-Derived Stromal Cells Arterioscler. Thromb. Vasc. Biol., December 1, 2005; 25(12): 2542 - 2547. [Abstract] [Full Text] [PDF]
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 M. Jinnin, H. Ihn, Y. Mimura, Y. Asano, K. Yamane, and K. Tamaki Matrix metalloproteinase-1 up-regulation by hepatocyte growth factor in human dermal fibroblasts via ERK signaling pathway involves Ets1 and Fli1 Nucleic Acids Res., June 21, 2005; 33(11): 3540 - 3549. [Abstract] [Full Text] [PDF]
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B. Xu, A. Bhattacharjee, B. Roy, G. M. Feldman, and M. K. Cathcart Role of Protein Kinase C Isoforms in the Regulation of Interleukin-13-induced 15-Lipoxygenase Gene Expression in Human Monocytes J. Biol. Chem., April 16, 2004; 279(16): 15954 - 15960. [Abstract] [Full Text] [PDF]
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 J. M. Ueland, J. Gwira, Z.-X. Liu, and L. G. Cantley The chemokine KC regulates HGF-stimulated epithelial cell morphogenesis Am J Physiol Renal Physiol, March 1, 2004; 286(3): F581 - F589. [Abstract] [Full Text]
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C.-W. Ni, H.-J. Hsieh, Y.-J. Chao, and D. L. Wang Shear Flow Attenuates Serum-induced STAT3 Activation in Endothelial Cells J. Biol. Chem., May 23, 2003; 278(22): 19702 - 19708. [Abstract] [Full Text] [PDF]
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 J. Rehman, R. V. Considine, J. E. Bovenkerk, J. Li, C. A. Slavens, R. M. Jones, and K. L. March Obesity is associated with increased levels of circulating hepatocyte growth factor J. Am. Coll. Cardiol., April 16, 2003; 41(8): 1408 - 1413. [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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 M. Sata and R. Nagai Phosphatidylinositol 3-Kinase: A Key Regulator of Vascular Tone? Circ. Res., August 23, 2002; 91(4): 273 - 275. [Full Text] [PDF]
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Q. Zeng, S. Chen, Z. You, F. Yang, T. E. Carey, D. Saims, and C.-Y. Wang Hepatocyte Growth Factor Inhibits Anoikis in Head and Neck Squamous Cell Carcinoma Cells by Activation of ERK and Akt Signaling Independent of NFkappa B J. Biol. Chem., July 5, 2002; 277(28): 25203 - 25208. [Abstract] [Full Text] [PDF]
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M. Torbenson, S. Q. Yang, H. Z. Liu, J. Huang, W. Gage, and A. M. Diehl STAT-3 Overexpression and p21 Up-Regulation Accompany Impaired Regeneration of Fatty Livers Am. J. Pathol., July 1, 2002; 161(1): 155 - 161. [Abstract] [Full Text] [PDF]
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I. Shiojima and K. Walsh Role of Akt Signaling in Vascular Homeostasis and Angiogenesis Circ. Res., June 28, 2002; 90(12): 1243 - 1250. [Abstract] [Full Text] [PDF]
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H. Nakagami, T.-X. Cui, M. Iwai, T. Shiuchi, Y. Takeda-Matsubara, L. Wu, and M. Horiuchi Tumor Necrosis Factor-{alpha} Inhibits Growth Factor-Mediated Cell Proliferation Through SHP-1 Activation in Endothelial Cells Arterioscler. Thromb. Vasc. Biol., February 1, 2002; 22(2): 238 - 242. [Abstract] [Full Text] [PDF]
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N. Wajih, J. Walter, and D. C. Sane Vascular Origin of a Soluble Truncated Form of the Hepatocyte Growth Factor Receptor (c-met) Circ. Res., January 11, 2002; 90(1): 46 - 52. [Abstract] [Full Text] [PDF]
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