| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Advances in Brief |
Department of Anatomy and Cell Biology [A. R. H., E. A. S., L. M. G. G., R. E. B. S., M. J. C. H.] and Holden Comprehensive Cancer Center, at the University of Iowa [E. A. S., G. B. S., R. E. B. S., M. J. C. H.], University of Iowa College of Medicine and the Dows Institute for Dental Research and Department of Prosthodontics [G. B. S.], University of Iowa College of Dentistry, Iowa City, Iowa, 52242-1109, and Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana 47907-1246 [K. L. C. K., M. S. K.]
 |
ABSTRACT |
|---|
 Top ABSTRACT IntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
|
Introduction |
|---|
|
TopABSTRACTIntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
Microarray analysis of the aggressive melanoma cell lines revealed differential expression of various protein tyrosine kinases as compared with the poorly aggressive melanoma cell lines (4 , 6) . Signal transduction pathways, consisting of PTKs,3 which are largely responsible for relaying signals through the cell by way of phosphorylation events, play critical and diverse roles in regulating cell adhesion, migration, and proliferation. The increased expression of various protein tyrosine kinases expressed by the aggressive melanoma cells prompted us to determine whether they may play a role in mediating vasculogenic mimicry.
In this study, we tested the hypothesis that melanoma cells that express a specific profile of tyrosine phosphorylated proteins would be facilitated in their ability to engage in vasculogenic mimicry, as determined by the formation of patterned tubular networks in vitro. Immunofluorescence staining of the aggressive melanoma cells grown on three-dimensional matrices indicated that areas of the cultures containing patterned tubular networks also showed high levels of tyrosine phosphorylation. The addition of general inhibitors of PTKs resulted in the inability of the aggressive melanoma cells to form the tubular networks. Further molecular analysis revealed Eck (Eck/EphA2) as one receptor PTK that was expressed and phosphorylated exclusively in the aggressive metastatic uveal melanoma tumor cells. Transient knockout of EphA2 expression in the aggressive uveal melanoma tumor cells resulted in the inhibition of tubular network formation by these cells. Together, these results demonstrate that PTKs, particularly EphA2, play a critical role in the formation of matrix-rich tubular networks by aggressive melanoma cells and may constitute one of the essential molecular determinants of vasculogenic mimicry demonstrated by melanoma.
|
Materials and Methods |
|---|
|
TopABSTRACTIntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
Three-Dimensional Cultures.
Fifteen µl of type I collagen (3.02 mg/ml; Collaborative Biomedical, Bedford, MA) were dropped onto 18-mm no. 1 glass coverslips in 12-well tissue culture plates. Forty-five µl of 100% ethanol were dropped onto each coverslip, and the collagen was allowed to polymerize for 5 min at room temperature. Forty-five µl of 1x PBS were dropped onto each coverslip, and the collagen was allowed to polymerize for an additional 5 min at room temperature. Five x 105 tumor cells were then seeded on top of the collagen gel. For experiments with herbimycin A, cells were allowed to incubate on the collagen for 3 h, after which the medium was removed and replaced with fresh medium containing herbimycin A (final concentration, 875 nM; Life Technologies, Inc.). Fresh medium containing herbimycin A was added to these cultures every 24 h for 4 days. Cells were determined to be viable by use of the trypan blue exclusion test.
Immunoprecipitation, Electrophoresis, and Immunoblotting.
Fifty µl of type I collagen (3.02 mg/ml; Collaborative Biomedical) were dropped into each well of a six-well tissue culture plate. One hundred and fifty µl of 100% ethanol were added to each well, and the collagen was allowed to polymerize for 5 min at room temperature. One hundred and fifty µl of 1x PBS were added to each well, and the collagen was allowed to continue polymerizing for an additional 5 min at room temperature. Two x 106 tumor cells were then seeded into each well. At the appropriate time points, cells plus the matrix were removed from the dishes by cell scraping. The medium, matrix, and cells were then centrifuged at 12,500 rpm for 5 min at 4°C. The pellets were washed once with ice-cold 1x PBS and then centrifuged at 12,500 rpm for 5 min at 4°C. The pellets were lysed with 1x RIPA buffer (50 mM Tris, 150 mM NaCl, 1% Triton X-100, and 0.5% deoxycholate) containing 25 µg/ml leupeptin (Sigma Chemical Co., St. Louis, MO), 10 µg/ml aprotinin (Sigma Chemical Co.), 1 mM sodium orthovanadate, and 2 mM EDTA and then centrifugation at 12,500 rpm for 5 min. The protein concentration of the samples was determined using a BCA Protein Assay Reagent kit (Pierce Corp., Rockford, IL), and whole cell lysates were analyzed by 10% SDS-PAGE and stained with Coomassie BBR-250 (Sigma Chemical Co.) to ensure equal loading (data not shown). Immunoprecipitations were preformed by continuous shaking for 1 h at 4°C with 5 µg of primary antibody/reaction, followed by 1-h incubation in the presence of protein A-Sepharose beads (15 µl; Sigma Chemical Co.). The protein A-Sepharose was washed thoroughly with RIPA buffer, and the protein was recovered by boiling in electrophoresis SDS sample buffer. Samples were separated by 10% SDS-PAGE and transferred to nitrocellulose (Schleicher and Schuell, Keene, NH.). Blots were blocked overnight at 4°C and for 1 h at room temperature in Tris-buffered saline (TBS) containing 0.05% Tween 20 (TBSTB), 0.1% BSA, and 2% cold fish gelatin (Sigma Chemical Co.). Blots were incubated with either anti-phosphotyrosine antibody (1:500 PY-20; Transduction Laboratories, San Diego, CA) or anti-EphA2 antibody (1 µg/ml; Upstate Biotechnology, Lake Placid, NY) for 1 h at room temperature with agitation, followed by incubation with a horseradish peroxidase-conjugated antimouse secondary antibody (1:5000; The Jackson Laboratory, Bar Harbor, ME). Blots were developed using an enhanced chemiluminescence detection kit (ECL; Amersham Pharmacia Biotech, Piscataway, NJ).
Immunocytochemistry.
Coverslips containing three-dimensional cultures were fixed with 3.7% formaldehyde in PBS for 10 min and then stored in TBS (50 mM Tris-HCl, 150 mM NaCl, and 1% sodium azide) at 4°C. Prior to staining, coverslips were rinsed with TBS. Coverslips where dipped in a 0.5% Triton X-100 solution for 6 min, followed by a rinse with TBS for 5 min. Coverslips where incubated for 1 h with anti-phosphotyrosine antibody (1:50 PY-20; Transduction Laboratories) and Texas Red-conjugated phalloidin (1:50; Molecular Probes, Eugene, OR) at room temperature. After incubation, the coverslips were rinsed with TBS for 5 min. The coverslips were then incubated for 1 h with a FITC-conjugated antimouse IgG (1:50; Cappel ICN, Irvine, CA) at room temperature and subsequently rinsed with TBS for 5 min. The coverslips were then mounted on glass slides using permanent mounting media.
For cross-sectional analysis of these three-dimensional cultures, transwells (Fisher Scientific) containing three-dimensional cultures of MUM-2B cells were frozen and sectioned at 7 µm. Slides were acetone fixed for 5 min, rinsed in PBS, and stained with anti-EphA2 kinase antibodies (1:50; Upstate Biotechnology), using LSAB plus alkaline phosphatase kit (Dako, Carpinteria, CA). Sections were then exposed to Vector red chromogen (Vector Laboratories, Burlingame, CA) for up to 5–7 min, rinsed in distilled water, counterstained with methyl green (0.5% aqueous purified with chloroform; Poly Scientific, Bay Shore, NY) for 10 min, and coverslipped with permanent mounting medium.
To help visualize the extracellular matrix-rich tubular networks within the three-dimensional cultures prepared on glass coverslips, the coverslips were stained with PAS, omitting hematoxylin counterstaining to reduce visual noise; black and white photography with a green filter was also used to enhance the PAS-positive patterns. Pictures were taken using an Axioskop 2 (Carl Zeiss, Inc., Thornwood, NY) and Spot camera (Diagnostic Instrument, Inc., Sterling Heights, MI) using the Zeiss Axiovision 2.0.5 software (Carl Zeiss, Inc.).
Transient Transfections.
To knock out the expression of EphA2 in the aggressive MUM-2B uveal melanoma cells, they where transiently transfected with phosphorothioate-modified antisense oligonucleotides (5'-CCAGCAGTACCGCTTCCTTGCCCTGCGGCCG-3'), inverted antisense oligonucleotides (5'-GCCGCGTCCCGTTCCTTCACCATGACGACC-3'), or untransfected to serve as a vehicle control. The cells were transfected with 8 µg of DNA using Lipofectamine PLUS Reagent (Life Technologies, Inc.), following the manufacturer’s protocol. Twenty-four h after transfection, the cells were harvested and seeded onto three-dimensional cultures consisting of type I collagen. After 48 h, the cultures were analyzed by bright-field microscopy for the presence of tubular networks using an Olympus IX-70 inverted microscope. Images were captured onto 35 mm Kodak T-Max professional film using an Olympus OM-4T camera. At 0 and 24 h, the treated cells were harvested from the three-dimensional matrix, and a Western blot was prepared as described previously (8)
. The Western blot was probed with anti-EphA2 antibodies and with anti-paxillin antibodies (a generous gift from Keith Burridge, University of North Carolina, Chapel Hill, NC) to serve as an internal loading control.
|
Results |
|---|
|
TopABSTRACTIntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
, on days 1, 3, and 6, respectively. However, the poorly aggressive MUM-2C uveal melanoma cell line is unable to form tubular networks under the same culture conditions, as depicted in Fig. 1, B, D, and F
, during days 1, 3, and 6, respectively. Light microscopic analysis of cross-sections of the patterned networks formed by MUM-2B cells demonstrates the hollow lumen-like structures that exist within the tubular networks (Fig. 1E
, inset).
|
, and the corresponding dual-fluorescence image is shown in Fig. 2C
, which delineated an area of intense phosphotyrosine staining associated with the tumor formed tubular networks. Actin staining highlights the cellular components contained within the spheroids of tumor cells surrounded by the tubular networks. In contrast, when the poorly aggressive MUM-2C cells were cultured on a three-dimensional matrix, they were unable to form tubular networks, as shown in Fig. 2B
; the fluorescence image from the same field (shown in Fig. 2D
) revealed dispersed phosphotyrosine staining of the MUM-2C cells, along with a lack of organization into patterned tubular networks as seen with the aggressive MUM-2B cells.
|
, which suggested that tyrosine phosphorylation may be associated with tubular network formation, we next examined whether inhibiting tyrosine kinases could affect tubular network formation by the aggressive MUM-2B uveal melanoma cells. Consequently, MUM-2B cells were cultured on a three-dimensional matrix in the presence or absence of 875 nM herbimycin A. Under these conditions, cells can be cultured up to 96 h without toxic effects (9)
. After 4 days in vitro, the aggressive MUM-2B uveal melanoma cells cultured in the absence of herbimycin A were able to form tubular networks as expected (Fig. 3A)
. However, the herbimycin A-treated MUM-2B cells were inhibited in their ability to form complete tubular networks; instead, clusters of cells and extracellular matrix were observed in the absence of organized networks, as demonstrated by PAS histochemistry (Fig. 3B)
. These cells were viable based on the trypan blue exclusion test (data not shown). Removal of the herbimycin A allowed the MUM-2B cells to recover and resume the formation of tubular networks within 4 additional days (Fig. 3C)
.
|
. Whole cell lysates were analyzed by 10% SDS-PAGE and stained with Coomassie blue to ensure equal loading (data not shown). The most significant difference appears to be a protein in the Mr 130,000 range that is phosphorylated in the MUM-2B cells but not in the MUM-2C cells.
|
. A Northern blot analysis verified that EphA2 was expressed exclusively in MUM-2B cells (data not shown), which was confirmed by Western blot analysis of an anti-EphA2 immunoprecipitation with these lysates (Fig. 4C)
. Furthermore, light microscopic immunohistochemical analysis of three-dimensional cultures of MUM-2B vasculogenic networks showed positive staining with anti-EphA2 antibodies (Fig. 4D)
, compared with no staining in the control tissues (Fig. 4E)
. These results suggest that phosphorylation of EphA2 may play a critical role in the aggressive MUM-2B uveal melanoma cells ability to form tubular networks and engage in vasculogenic mimicry.
To directly test the role of EphA2 in the process of tubular network formation, antisense oligonucleotides were used to transiently diminish EphA2 expression in the aggressive uveal melanoma cells. The treated cells were then assessed for their ability to develop tubular structures when cultured on a three-dimensional matrix. Light microscopic analysis of three-dimensional cultures indicated that 48 h after transfection, the cells that were treated with the inverted antisense oligonucleotides were able to form tubular networks as shown in Fig. 5A
, whereas the cells treated with the EphA2 antisense oligonucleotides were inhibited in their ability to initiate tubular networks (Fig. 5B)
. Western blot analysis confirmed that antisense oligonucleotide treatment of aggressive melanoma cells decreased EphA2 expression (Fig. 5C)
. Levels of paxillin were assessed as relatively unchanged in the control and experimental samples. These results implicate the importance of EphA2 expression in the ability of the aggressive uveal melanoma cell to engage in tubular network formation in vitro.
|
|
Discussion |
|---|
|
TopABSTRACTIntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
Epithelial cell kinase is a member of the Eph family of protein receptor tyrosine kinases. Eph is a large family that currently consists of 14 members (10) . The EphA2 receptor tyrosine kinase has a molecular weight of Mr 130,000 and contains 976 amino acids (11) . The cytoplasmic domain contains the tyrosine catalytic domain of the protein and a putative phosphorylation site at Y-772 (11) . EphA2 is normally expressed in tissues that contain a high proportion of epithelial cells such as skin, intestine, lung, ovary, and kidney (11, 12, 13) . The ligand for EphA2 is ephrin-A1 (otherwise known as B61; Ref. 14 , 15 ). Binding of ephrin-A1 causes EphA2 to become phosphorylated; however, it has been found that EphA2 can also be constitutively phosphorylated in unstimulated cells (12) . Early experiments using a yeast two-hybrid system showed that EphA2 was able to bind to the COOH-terminal SH2 domain of the p85 subunit of PI3K, and activation of EphA2 by ephrin-A1 then stimulated activation of PI3K (16) . Also, using the same yeast two-hybrid system, a novel protein containing SH3 and SH2 adapter modules was found to bind to activated EphA2 and was named src-like adapter protein (17) . Recently, it was revealed that the nonactivated form of EphA2 was able to associate with FAK in vivo, and the complex becomes disrupted, leading to the dephosphorylation of FAK and paxillin upon activation of EphA2 by ephrin-A1 (18) . These results demonstrated decreases in integrin-mediated cell adhesion, spreading, and migration (8 , 18) . Lastly, the dephosphorylation of FAK by activation of EphA2 could be suppressed by addition of inhibitors to protein tyrosine phosphatases, suggesting that activated EphA2 may recruit various phosphatases.
EphA2 is not normally expressed in normal melanocytes but is often up-regulated in various primary melanoma tumors as well as those from metastatic lesions (4 , 19, 20, 21, 22) . The role of EphA2 in tumor progression remains largely unknown. Studies have implicated EphA2 to play a role in the growth and proliferation of EphA2 expressing melanoma cells, as well as induction of vascularization in advanced melanoma (22) , thus underscoring its importance in tumor progression.
Our results validated the hypothesis that melanoma cells expressing a specific profile of tyrosine phosphorylated proteins would be facilitated in their ability to participate in vasculogenic mimicry, the formation of patterned tubular networks in vitro, and suggests that protein tyrosine kinases leading to phosphorylation of various signal transduction proteins are also important for the process of vasculogenic mimicry. We have identified one such receptor protein tyrosine kinase, EphA2, as being exclusively expressed and phosphorylated in the highly aggressive uveal melanoma cell line, MUM-2B, but not in the poorly aggressive MUM-2C cell line. These data extend the molecular microarray analysis of these two cell lines, isolated from the same patient, which demonstrated a genetic instability or deregulation in the aggressive cells that allowed them to indiscriminately express genes that are associated with embryonic-like, pluripotent stem cells (4) , which may facilitate their ability to engage in vasculogenic mimicry and form patterned tubular networks.
The signaling pathways downstream of EphA2 need to be further examined to discern its precise role in vasculogenic mimicry. Interestingly, it has been reported that kidney epithelial cells expressing elevated levels of PI3K activity are able to form branching tubules when cultured on a three-dimensional matrix composed of type I collagen (23) . On the basis of these observations and ours, it is tempting to speculate that activated EphA2 could be signaling through PI3K in the aggressive uveal melanoma cells, thus resulting in the formation of tubular networks in three-dimensional matrices.
Our observations reveal for the first time that epithelial cell kinase is a critical molecular determinant for vasculogenic mimicry in aggressive melanoma cells. Inhibition of tyrosine phosphorylation abrogates the ability of the aggressive melanoma cells to form embryonic-like patterns of vasculogenic networks, which may have future implications for clinical use in treating uveal melanoma tumors containing patterned, extracellular matrix-rich networks surrounding spheroids of tumor cells. Not only have these patterned networks been found during embryogenesis and in the formation of the primitive microcirculation (24, 25, 26) , but most recently they have been observed in Drosophila tumors containing mutations in the lats gene (large tumor suppressor gene), a model with no blood flow or endothelial cells (27 , 28) . Although the function of these networks is not completely understood, it is plausible that they may be important in tumor perfusion, dissemination, and drug resistance. Further investigation of Eck may help to elucidate the molecular regulation of vasculogenic mimicry and may contribute new therapeutic targets for clinical intervention in aggressive melanoma tumors.
|
ACKNOWLEDGMENTS |
|---|
|
FOOTNOTES |
|---|
1 This research was funded by NIH Grants CA59702 and CA80318 (to M. J. C. H.), CA83137 (to R. E. B. S.), and United States Army Medical Research and Materiel Command DAMD17-98–1-8146 (to M. S. K.). 
2 To whom requests for reprints should be addressed, at Department of Anatomy and Cell Biology, 51 Newton Road, 1–100 Bowen Science Building, University of Iowa, Iowa City, IA 52242-1109. Phone: (319) 335-7755; Fax (319) 335-7770; E-mail: mary-hendrix{at}uiowa.edu
3 The abbreviations used are: PTK, protein tyrosine kinase; Eck/EphA2, epithelial cell kinase; PAS, periodic acid-Schiff; FAK, focal adhesion kinase; PI3K, phosphoinositide 3-kinase.
Received 6/ 1/00. Accepted 2/28/01.
|
REFERENCES |
|---|
|
TopABSTRACTIntroductionMaterials and MethodsResultsDiscussionREFERENCES |
|---|
-induced angiogenesis. Science (Wash. DC), 286: 567-569, 1995.
This article has been cited by other articles:
|
|
 |
|
  M. Parri, M. L. Taddei, F. Bianchini, L. Calorini, and P. Chiarugi EphA2 Reexpression Prompts Invasion of Melanoma Cells Shifting from Mesenchymal to Amoeboid-like Motility Style Cancer Res., March 1, 2009; 69(5): 2072 - 2081. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-C. Lissitzky, D. Parriaux, E. Ristorcelli, A. Verine, D. Lombardo, and P. Verrando Cyclic AMP Signaling as a Mediator of Vasculogenic Mimicry in Aggressive Human Melanoma Cells In vitro Cancer Res., February 1, 2009; 69(3): 802 - 809. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Wykosky and W. Debinski The EphA2 Receptor and EphrinA1 Ligand in Solid Tumors: Function and Therapeutic Targeting Mol. Cancer Res., December 1, 2008; 6(12): 1795 - 1806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. A. Mourad-Zeidan, V. O. Melnikova, H. Wang, A. Raz, and M. Bar-Eli Expression Profiling of Galectin-3-Depleted Melanoma Cells Reveals its Major Role in Melanoma Cell Plasticity and Vasculogenic Mimicry Am. J. Pathol., December 1, 2008; 173(6): 1839 - 1852. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Buettner, T. Mesa, A. Vultur, F. Lee, and R. Jove Inhibition of Src Family Kinases with Dasatinib Blocks Migration and Invasion of Human Melanoma Cells Mol. Cancer Res., November 1, 2008; 6(11): 1766 - 1774. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Noberini, M. Koolpe, S. Peddibhotla, R. Dahl, Y. Su, N. D. P. Cosford, G. P. Roth, and E. B. Pasquale Small Molecules Can Selectively Inhibit Ephrin Binding to the EphA4 and EphA2 Receptors J. Biol. Chem., October 24, 2008; 283(43): 29461 - 29472. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. P. Robertson Mig-7 Linked to Vasculogenic Mimicry Am. J. Pathol., May 1, 2007; 170(5): 1454 - 1456. [Full Text] [PDF]
|
|
|
|
|
|
A. P. Petty, K. L. Garman, V. D. Winn, C. M. Spidel, and J. S. Lindsey Overexpression of Carcinoma and Embryonic Cytotrophoblast Cell-Specific Mig-7 Induces Invasion and Vessel-Like Structure Formation Am. J. Pathol., May 1, 2007; 170(5): 1763 - 1780. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N.-Y. Yang, E. B. Pasquale, L. B. Owen, and I. M. Ethell The EphB4 Receptor-tyrosine Kinase Promotes the Migration of Melanoma Cells through Rho-mediated Actin Cytoskeleton Reorganization J. Biol. Chem., October 27, 2006; 281(43): 32574 - 32586. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. R. Hess, L.-M. Postovit, N. V. Margaryan, E. A. Seftor, G. B. Schneider, R. E.B. Seftor, B. J. Nickoloff, and M. J.C. Hendrix Focal Adhesion Kinase Promotes the Aggressive Melanoma Phenotype Cancer Res., November 1, 2005; 65(21): 9851 - 9860. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Hu, K. L. Carles-Kinch, D. P. Zelinski, and M. S. Kinch EphA2 Induction of Fibronectin Creates a Permissive Microenvironment for Malignant Cells Mol. Cancer Res., October 1, 2004; 2(10): 533 - 540. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. H. Thaker, M. Deavers, J. Celestino, A. Thornton, M. S. Fletcher, C. N. Landen, M. S. Kinch, P. A. Kiener, and A. K. Sood EphA2 Expression Is Associated with Aggressive Features in Ovarian Carcinoma Clin. Cancer Res., August 1, 2004; 10(15): 5145 - 5150. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. A. Scott, J. K. Vass, E. K. Parkinson, D. A. F. Gillespie, J. N. Winnie, and B. W. Ozanne Invasion of Normal Human Fibroblasts Induced by v-Fos Is Independent of Proliferation, Immortalization, and the Tumor Suppressors p16INK4a and p53 Mol. Cell. Biol., February 15, 2004; 24(4): 1540 - 1559. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Dobrzanski, K. Hunter, S. Jones-Bolin, H. Chang, C. Robinson, S. Pritchard, H. Zhao, and B. Ruggeri Antiangiogenic and Antitumor Efficacy of EphA2 Receptor Antagonist Cancer Res., February 1, 2004; 64(3): 910 - 919. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. M. S. Alves, O. Faure, S. Graff-Dubois, D.-A. Gross, S. Cornet, S. Chouaib, I. Miconnet, F. A. Lemonnier, and K. Kosmatopoulos EphA2 as Target of Anticancer Immunotherapy: Identification of HLA-A*0201-Restricted Epitopes Cancer Res., December 1, 2003; 63(23): 8476 - 8480. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. T. Coffman, M. Hu, K. Carles-Kinch, D. Tice, N. Donacki, K. Munyon, G. Kifle, R. Woods, S. Langermann, P. A. Kiener, et al. Differential EphA2 Epitope Display on Normal versus Malignant Cells Cancer Res., November 15, 2003; 63(22): 7907 - 7912. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kunz, S. Moeller, D. Koczan, P. Lorenz, R. H. Wenger, M. O. Glocker, H.-J. Thiesen, G. Gross, and S. M. Ibrahim Mechanisms of Hypoxic Gene Regulation of Angiogenesis Factor Cyr61 in Melanoma Cells J. Biol. Chem., November 14, 2003; 278(46): 45651 - 45660. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Ruf, E. A. Seftor, R. J. Petrovan, R. M. Weiss, L. M. Gruman, N. V. Margaryan, R. E. B. Seftor, Y. Miyagi, and M. J. C. Hendrix Differential Role of Tissue Factor Pathway Inhibitors 1 and 2 in Melanoma Vasculogenic Mimicry Cancer Res., September 1, 2003; 63(17): 5381 - 5389. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. R. Hess, E. A. Seftor, R. E. B. Seftor, and M. J. C. Hendrix Phosphoinositide 3-Kinase Regulates Membrane Type 1-Matrix Metalloproteinase (MMP) and MMP-2 Activity during Melanoma Cell Vasculogenic Mimicry Cancer Res., August 15, 2003; 63(16): 4757 - 4762. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Lu, K. D. Miller, Y. Gokmen-Polar, M.-H. Jeng, and M. S. Kinch EphA2 Overexpression Decreases Estrogen Dependence and Tamoxifen Sensitivity Cancer Res., June 15, 2003; 63(12): 3425 - 3429. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Walker-Daniels, A. R. Hess, M. J.C. Hendrix, and M. S. Kinch Differential Regulation of EphA2 in Normal and Malignant Cells Am. J. Pathol., April 1, 2003; 162(4): 1037 - 1042. [Full Text] [PDF]
|
|
|
|
|
|
M. S. Kinch, M.-B. Moore, and D. H. Harpole Jr. Predictive Value of the EphA2 Receptor Tyrosine Kinase in Lung Cancer Recurrence and Survival Clin. Cancer Res., February 1, 2003; 9(2): 613 - 618. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. D. Miller, C. J. Sweeney, and G. W. Sledge The Snark is a Boojum: the continuing problem of drug resistance in the antiangiogenic era Ann. Onc., January 1, 2003; 14(1): 20 - 28. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Cohen, A. Zavala-Pompa, J. H. Sequeira, M. Shoji, D. G. Sexton, G. Cotsonis, F. Cerimele, B. Govindarajan, N. Macaron, and J. L. Arbiser Mitogen-actived Protein Kinase Activation Is an Early Event in Melanoma Progression Clin. Cancer Res., December 1, 2002; 8(12): 3728 - 3733. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Koolpe, M. Dail, and E. B. Pasquale An Ephrin Mimetic Peptide That Selectively Targets the EphA2 Receptor J. Biol. Chem., November 27, 2002; 277(49): 46974 - 46979. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Cheng, D. M. Brantley, H. Liu, Q. Lin, M. Enriquez, N. Gale, G. Yancopoulos, D. P. Cerretti, T. O. Daniel, and J. Chen Blockade of EphA Receptor Tyrosine Kinase Activation Inhibits Vascular Endothelial Cell Growth Factor-Induced Angiogenesis Mol. Cancer Res., November 1, 2002; 1(1): 2 - 11. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Herlyn, M. Padarathsingh, L. Chin, M. Hendrix, D. Becker, M. Nelson, Y. DeClerck, J. McCarthy, and S. Mohla New Approaches to the Biology of Melanoma: A Workshop of the National Institutes of Health Pathology B Study Section Am. J. Pathol., November 1, 2002; 161(5): 1949 - 1957. [Full Text] [PDF]
|
|
|
|
|
|
K. D. Kikawa, D. R. Vidale, R. L. Van Etten, and M. S. Kinch Regulation of the EphA2 Kinase by the Low Molecular Weight Tyrosine Phosphatase Induces Transformation J. Biol. Chem., October 11, 2002; 277(42): 39274 - 39279. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Liu, H. Huang, F. Donate, C. Dickinson, R. Santucci, A. El-Sheikh, R. Vessella, and T. S. Edgington Prostate-specific Membrane Antigen Directed Selective Thrombotic Infarction of Tumors Cancer Res., October 1, 2002; 62(19): 5470 - 5475. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Carles-Kinch, K. E. Kilpatrick, J. C. Stewart, and M. S. Kinch Antibody Targeting of the EphA2 Tyrosine Kinase Inhibits Malignant Cell Behavior Cancer Res., May 1, 2002; 62(10): 2840 - 2847. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
M. J. C. Hendrix, R. E. B. Seftor, E. A. Seftor, L. M. Gruman, L. M. L. Lee, B. J. Nickoloff, L. Miele, D. D. Sheriff, and G. C. Schatteman Transendothelial Function of Human Metastatic Melanoma Cells: Role of the Microenvironment in Cell-Fate Determination Cancer Res., February 1, 2002; 62(3): 665 - 668. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Shirakawa, H. Kobayashi, Y. Heike, S. Kawamoto, M. W. Brechbiel, F. Kasumi, T. Iwanaga, F. Konishi, M. Terada, and H. Wakasugi Hemodynamics in Vasculogenic Mimicry and Angiogenesis of Inflammatory Breast Cancer Xenograft Cancer Res., January 1, 2002; 62(2): 560 - 566. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. D. Zantek, J. Walker-Daniels, J. Stewart, R. K. Hansen, D. Robinson, H. Miao, B. Wang, H.-J. Kung, M. J. Bissell, and M. S. Kinch MCF-10A-NeoST: A New Cell System for Studying Cell-ECM and Cell-Cell Interactions in Breast Cancer Clin. Cancer Res., November 1, 2001; 7(11): 3640 - 3648. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. E. B. Seftor, E. A. Seftor, N. Koshikawa, P. S. Meltzer, L. M. G. Gardner, M. Bilban, W. G. Stetler-Stevenson, V. Quaranta, and M. J. C. Hendrix Cooperative Interactions of Laminin 5 {gamma}2 Chain, Matrix Metalloproteinase-2, and Membrane Type-1-Matrix/Metalloproteinase Are Required for Mimicry of Embryonic Vasculogenesis by Aggressive Melanoma Cancer Res., September 1, 2001; 61(17): 6322 - 6327. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Cancer Research | Clinical Cancer Research |
| Cancer Epidemiology Biomarkers & Prevention | Molecular Cancer Therapeutics |
| Molecular Cancer Research | Cancer Prevention Research |
| Cancer Prevention Journals Portal | Cancer Reviews Online |
| Annual Meeting Education Book | Meeting Abstracts Online |
