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Tumor Biology |
Departments of Basic Medical Sciences [D. P. Z., N. D. Z., J. C. S., M. S. K.] and Veterinary Pathobiology [A. R. I.], Purdue University, West Lafayette, Indiana 47907-1246
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ABSTRACT |
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INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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When malignant cells metastasize to distant sites in the body, morbidity and mortality increase significantly (3) . Metastatic cells have acquired the abilities to break away from the primary tumor, translocate to distant sites in the body, and colonize a foreign microenvironment (4) . At the cellular level, malignant cells have overcome restraints on cell growth and migration that result from physical linkages and signals conveyed by cell-cell contacts (5) . Malignant cells often have increased interactions with surrounding ECM3 proteins, which provide linkages and signals that promote several aspects of metastasis (6) .
Our previous studies revealed that the levels of protein tyrosine phosphorylation regulate a balance between cell-cell and cell-ECM adhesions in epithelial cells (7) . Using oncogene-transformed mammary epithelial cells, we showed that elevated tyrosine kinase activity weakens cell-cell contacts and promotes ECM adhesions (7) . To identify tyrosine kinases that control tumor cell adhesion, we developed novel technologies to generate monoclonal antibodies against tyrosine kinases in cancer cells (8) . We focused on one particular antigen that was functionally altered in oncogene-transformed epithelial cells. This antigen was identified as EphA2 (ECK). EphA2 is a Mr 130,000 receptor tyrosine kinase that is expressed on adult epithelia (9) , where it is found at low levels and enriched within sites of cell-cell adhesion (10) . The subcellular localization is important because EphA2 binds five different ligands, ephrinA1–5, which are attached to the cell membrane (11) .
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Western Blot Analysis and Immunoprecipitation.
Western blot analyses were performed as described previously (10)
, and antibody binding was detected by enhanced chemiluminescence (Pierce, Rockford, IL) and autoradiography (Kodak X-OMAT; Kodak, Rochester, NY). To confirm equal sample loading, the blots were stripped and reprobed with antibodies specific for ß-catenin or vinculin.
Immunohistochemistry and Immunofluorescence Staining.
Formalin-fixed, paraffin-embedded "sausage" slides, each containing 15–30 breast cancer specimens (kindly provided by B. J. Kerns; BioGenex, San Ramon, CA), were stained and scored as described previously (12)
. Mean immunostaining intensity in benign and malignant breast was compared using Student’s t test with statistical software (SAS for Windows v.6.04 and Microsoft Excel ’97), defining P < 0.05 as significant. Staining of cell monolayers with EphA2 antibodies (clones D7 or B2D6) was performed as described previously (10)
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Transfection and Selection.
Monolayers of MCF-10A cells were cotransfected with the pNeoMSV-EphA2 (generously provided by Dr. T. Hunter, Scripps Institute, La Jolla, CA) and pBABE-Puro eukaryotic expression vectors, at a 4:1 ratio, using LipofectAMINE Plus (Life Technologies, Inc., Grand Island, NY). As a control for the transfection procedure, a parallel transfection was performed using pNeoMSV and pBABE-Puro. Puromycin-resistant cells were selected by supplementing the growth medium with 1 µg/ml puromycin (Sigma, St. Louis, MO). EphA2 overexpression was confirmed by Western blot analysis with specific antibodies. All experiments were performed using bulk culture transfectants, and identical results were obtained using cells from two separate transfections with EphA2 cDNAs. Parental cells and cultures transfected with pBABE-Puro were used as negative controls.
Colony Formation in Soft Agar.
Colony formation in soft agar was performed as described previously (13)
. Colony formation was scored microscopically, and clusters of at least three cells were defined as a positive result. For experiments with EA1, 0.5 µg/ml EA1 or a matched vehicle (50% glycerol in PBS) was included in top agar solution, and ligand was replenished daily with fresh media. The data shown are pooled from 10 separate high-power microscopic fields from each sample and representative of at least three separate experiments.
Cell Behavior in Matrigel.
The behavior of cells in Matrigel was analyzed as described previously (14)
. Briefly, tissue culture dishes were coated with Matrigel (Collaborative, Bedford, MA) at 37°C before adding 1 x 105 vector- or EphA2-transfected MCF-10A cells. The behavior of EphA2-overexpressing cells was assessed at 6-h intervals using an inverted light microscope (Olympus IX-70). For experiments with EA1, the culture medium was supplemented with 0.5 µg/ml EA1 or an appropriately matched vehicle control. All images were recorded onto 35-mm film (T-Max-400; Kodak, Rochester, NY).
Xenograft Analyses.
Athymic (nu/nu) 3–4-week-old mice were purchased from Harlan Sprague Dawley (Indianapolis, IN) and Charles River (Wilmington, MA) and acclimated for 7–10 days. For s.c. implantation, 1 x 106 or 5 x 106 vector- or EphA2-transfected MCF-10A cells were suspended in 100 µl of fresh media and injected into the right craniolateral thorax (axilla) using a 23-gauge needle. For tail vein injections, 1 x 106 cells were injected into the tail vein, and mice were monitored for 7–28 days. At necropsy, primary tumors and all organs were evaluated macroscopically for the presence of tumors. Tissue samples of the primary tumor and organs were fixed in 10% buffered neutral formalin and embedded in paraffin. Tissue sections of the tumors and lung were stained with H&E to assess morphology. Lung sections were stained with antibodies specific for cytokeratin (AE1/AE3) or factor VIII-related antigen (DAKO, Carpinteria, CA) to confirm the epithelial nature of lung metastases.
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RESULTS |
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. Immunohistochemical staining of formalin-fixed, paraffin-embedded tissue sections revealed a low level of EphA2 immunoreactivity in benign mammary epithelia, with an average staining intensity of 0.1 (using a 0–3 scale to report staining intensity; Fig. 1B
). EphA2 immunoreactivity was increased in breast carcinoma specimens, with an average staining intensity of 2.9. Interestingly, EphA2 immunoreactivity in the breast carcinoma cells was diffusely distributed throughout the cytoplasm (Fig. 1A
, top). Increased staining intensity was accompanied by a larger percentage of carcinoma cells (an average of 87%) that stained positive for EphA2 as compared with benign mammary epithelial cells (an average of 3%).
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, Lanes 1–3), more EphA2 was detected in aggressive carcinoma cells (Fig. 2A
, Lanes 4–8). Identical results were obtained when equivalent numbers of cells or equal amounts of protein were analyzed (data not shown). Increased levels of EphA2 in aggressive cancer cell models were also confirmed using different EphA2 antibodies (D7, B2D6, and EK166B; data not shown), revealing that the differences in EphA2 levels did not reflect changes in a single epitope. The blots were stripped and reprobed with antibodies specific for ß-catenin (Fig. 2A)
or vinculin (data not shown), which confirmed equal sample loading.
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, top panel). After establishing cultures of MCF-10A cells with stable overexpression of EphA2, microscopic evaluation revealed differences in the cell morphology as compared with vector-transfected control cells (Fig. 2C)
. Nontransformed MCF-10A cells displayed an epithelial morphology and interacted with one another, even at low cell density. In contrast, EphA2-overexpressing MCF-10A cells (MCFEphA2 cells) adopted a fibroblast-like morphology and did not form cell-cell contacts, even at high cell density (data not shown). To confirm that the mesenchymal morphology did not represent clonal variation, a separate sample of MCF-10A cells was transfected with EphA2 cDNAs and yielded identical results.
EphA2 Overexpression Decreases Ligand-mediated Stimulation.
Because stable cell-cell contacts cause EphA2 to become enriched within sites of cell-cell contact (10)
, we assessed EphA2 subcellular localization by immunostaining with specific antibodies (Fig. 2C)
. The EphA2 on nontransformed MCF-10A cells was restricted to a narrow line where adjacent cells came into direct contact with each other, with little staining of membrane that was not in contact with neighboring cells. In contrast, the pattern of EphA2 staining on MCFEphA2 cells was diffuse, with little staining of cell-cell contacts. Notably, the cytoplasmic immunoreactivity of EphA2, which was prominent in tumor specimens, was also observed in MCFEphA2 cells.
The lack of EphA2 within the cell-cell contacts of MCFEphA2 cells was intriguing because EphA2 is stimulated by ligands that are anchored to the cell membrane (11
, 17)
. To measure EphA2 stimulation, the P-Tyr content of immunoprecipitated EphA2 was measured by Western blot analysis with P-Tyr-specific antibodies (Fig. 2B
, middle panel). Whereas the EphA2 in vector-transfected MCF-10A cells was tyrosine phosphorylated, EphA2 was not tyrosine phosphorylated in MCFEphA2 cells. The decreased P-Tyr content was confirmed using multiple EphA2 antibodies for immunoprecipitation (D7 and B2D6) and different P-Tyr-specific antibodies (4G10 and PY-20) for Western blot analyses (Table 1)
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, middle panel). Despite its inability to interact with its endogenous ligands, the EphA2 in MCFEphA2 cells could respond to exogenous stimuli.
EphA2 Overexpression Causes Malignant Transformation.
The pattern of defects in cell adhesion, EphA2 subcellular distribution, and P-Tyr content in MCFEphA2 cells were all reminiscent of metastatic cells (10)
, which prompted us to ask whether EphA2 overexpression induces malignant transformation. MCFEphA2 cells were found to colonize soft agar. Whereas vector-transfected MCF-10A cells formed fewer than 3 colonies/high-power field, MCFEphA2 cells displayed increased colony growth in soft agar, with an average of 30 colonies/sample (P < 3 x 10-7; Fig. 3A
). We then tested whether the decreased ligand binding in MCFEphA2 cells was related to colony formation in soft agar. To test this, MCFEphA2 cells were suspended in soft agar in the presence or absence of 0.5 µg/ml EA1. EA1 reduced colony formation in soft agar by 49% relative to vehicle-treated controls (P < 5 x 10-6). Thus, EphA2 stimulation reversed the effects of EphA2 overexpression.
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, left side). In contrast, MCFEphA2 cells adopted a stellate organization (Fig. 3B
, top right) that was indistinguishable from the behavior of aggressive breast cancer cells (e.g., MDA-MB-231 and MDA-MB-435; data not shown). To test whether EphA2 stimulation could alter cell behavior on Matrigel, the MCFEphA2 cells were treated with 0.5 µg/ml EA1, which restored a spherical phenotype that was comparable to that of nontransformed MCF-10A cells (Fig. 3B
, bottom right).
EphA2 Overexpression Confers Tumorigenic Potential.
Because in vitro analyses of transformation do not always predict tumorigenic potential in vivo, control or EphA2-overexpressing MCF-10A cells were implanted in athymic (nu/nu) mice (Fig. 4)
. The s.c. injection of MCFEphA2 cells caused the formation of palpable tumors within 4 days (Fig. 4, A–C)
in 19 of 19 mice. The median volume of resulting tumors was related to the number of implanted cells and reached an average of 300 mm3 (for samples injected with 5 x 106 cells) within 10 days (Fig. 4E)
. Necropsy revealed that the tumors were firmly attached to the underlying axillary muscle and surrounded by fibrous tissue (data not shown). Histologically, the neoplastic cells were invasive and associated with fibrous connective tissue (Fig. 4, B and C)
. These neoplastic cells exhibited moderate cytoplasmic and nuclear pleiomorphism and formed dysplastic tubular and secreting structures. In control experiments, cells transfected with vector DNA failed to grow in athymic mice (0 of 13 mice; Fig. 4E
), and necropsy failed to identify any growth or invasion of these cells (data not shown).
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. Immunohistochemical staining with cytokeratin antibodies confirmed the epithelial nature of the emboli, and a lack of anti-factor VIII-related antigen immunoreactivity revealed that the thrombus did not represent an abnormal or atypical outgrowth of endothelial cells (data not shown). No pulmonary emboli were observed in mice that had been injected with control MCF-10A cells (Fig. 4E)
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Consistent results with several cell models suggest that elevated levels of EphA2 are relevant to breast cancer. High levels of EphA2 are found in a large number of clinical specimens and aggressive cell models of breast cancer. Recent studies reveal that high levels of EphA2 may also be found in advanced melanoma (20) , colon cancer (21) , and prostate cancer (12) . The fact that elevated EphA2 levels are found on multiple types of cancer suggests that EphA2 overexpression may be a common event in the metastatic progression of carcinoma cells.
Our results provide the first evidence that EphA2 is not merely a marker but an active participant in tumorigenesis. EphA2-overexpressing MCF-10A cells displayed the hallmarks of malignant transformation as defined in vitro and in vivo. EphA2-transformed MCF-10A cells formed tumors in vivo at a high frequency, which is remarkable given that other oncogenes (e.g., Ras, HER2, and TC21) are insufficient to convey tumorigenic potential on MCF-10A cells (13 , 14) . Thus, we suggest that EphA2 overexpression may be particularly relevant to breast cancer.
EphA2 overexpression causes defects in cell-cell contacts that are characteristic of aggressive cancer cells. EphA2 weakens cell-cell contacts and thereby prevents EphA2 from interacting with its ligands, which are anchored to the surface of neighboring cells. Consistent with this, the highest levels of EphA2 are consistently found on tumor-derived breast cell lines that have weak cell-cell contacts (10 , 16) . Moreover, the EphA2 in these aggressive cancer cells is not tyrosine phosphorylated (10) . One possible explanation for the weakened cell-cell adhesions is that overexpressed EphA2 may phosphorylate adhesion or cytoskeletal proteins and thereby destabilize cell-cell adhesions. Consistent with this, elevated levels of protein tyrosine phosphorylation have been shown to destabilize cell-cell adhesions (7 , 22) . Further support is provided by evidence that EphA2 interacts with important adhesion and cytoskeletal proteins, including E-cadherin, Src-like adapter protein, and phosphatidylinositol 3'-kinase (10 , 18 , 23) . Another possibility is that EphA2 alters the expression of important adhesion molecules. Future studies will be needed to identify the molecular targets of EphA2 in malignant cells.
The weakened cell-cell adhesions of EphA2-overexpressing cells are notable because EphA2 binds a membrane-anchored ligand. We have recently shown that EphA2 in nontransformed epithelia is enriched within sites of cell-cell contact, where it interacts with ligand and becomes tyrosine phosphorylated (10)
. We demonstrate here that EphA2 overexpression causes EphA2 to become diffusely distributed. Consequently, the overexpressed EphA2 fails to interact with ligand and become tyrosine phosphorylated. Interestingly, EphA2 immunoreactivity in EphA2-overexpressing cells and in clinical specimens of breast cancer was similarly diffuse and cytoplasmic (see Fig. 1A
). The cytoplasmic localization of EphA2 contrasts with its known localization within sites of cell-cell contact between nontransformed epithelial cells (9)
. These results lead us to postulate that the levels of EphA2 protein influence its subcellular localization and thereby regulate ligand binding.
EphA2 overexpression causes malignant transformation and decreases ligand binding. These properties appear to be directly linked because EphA2 stimulation by soluble ligands reverses the malignant behavior of EphA2-transformed cells. Ligand-mediated tyrosine phosphorylation of EphA2 also decreases the growth and invasiveness of malignant breast and prostate cancer cells (10 , 18) . Thus, whereas ligand binding inhibits tumor cell growth, EphA2 overexpression causes malignant transformation and tumorigenesis. Taken together, these results indicate that the expression levels and ligand binding properties work together to allow EphA2 to differentially regulate tumor cell growth and invasiveness.
EphA2 overexpression may cause malignant transformation by regulating cell contact with the ECM. In contrast to evidence that ligand-mediated stimulation of EphA2 blocks ECM attachments (10 , 18) , ECM adhesions are increased in EphA2-transformed MCF-10A cells relative to nontransformed epithelial cells.4 Many different lines of investigation have shown that ECM adhesions provide linkages and signals that promote cell growth, migration, and survival (6 , 24 , 25) . The molecular basis by which EphA2 regulates ECM adhesions remains largely unknown. However, EphA2 has been shown to interact with a variety of cytoskeletal and signaling proteins, including phosphatidylinositol 3'-kinase, FAK, SHP-2, and a Src-like adapter protein (18 , 23 , 26) . These protein interactions are intriguing because each of the associated proteins has been independently found to regulate cell growth or ECM adhesion (24 , 27 , 28) .
Overexpressed receptor tyrosine kinases can facilitate new and efficacious modalities for targeted intervention against cancer cells (2)
. A recent success arose from antibody targeting of HER2, a receptor tyrosine kinase that is overexpressed on some breast cancer cells (2)
. Unfortunately, HER2 overexpression is limited to one-third of breast carcinomas and is sporadic on other tumor types, which underscores the need for new targets. Our results suggest that EphA2 might provide a target for intervention against aggressive breast cancers. At minimum, EphA2 overexpression may identify a larger or different set of tumors than HER2. Strong EphA2 immunoreactivity was detected in 5 of 12 (
40%) breast cancer specimens, whereas strong HER2 immunoreactivity was limited to 2 of 12 samples (data not shown). Our evidence suggests that strategies that restore or mimic the effects of ligand could negatively regulate tumor cell growth and invasiveness (10
, 18)
. This latter approach would redirect the function of an overexpressed oncoprotein so that it blocks tumor cell growth and invasiveness.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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1 Supported by the American Cancer Society, NIH, Department of Defense Breast Cancer Research Program, and the Howard Hughes Medical Institute (N. D. Z.). 
2 To whom requests for reprints should be addressed, at Department of Basic Medical Sciences, 1246 Lynn Hall, Purdue University, West Lafayette, Indiana 47907-1246.
3 The abbreviations used are: ECM, extracellular matrix; EA1, EphrinA1-Fc; P-Tyr, phosphotyrosine; 3dRBM, 3-dimensional, reconstituted basement membrane.
4 D. P. Zelinski and M. S. Kinch, unpublished results.
Received 8/10/00. Accepted 1/ 3/01.
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S. Abraham, D. W. Knapp, L. Cheng, P. W. Snyder, S. K. Mittal, D. S. Bangari, M. Kinch, L. Wu, J. Dhariwal, and S. I. Mohammed Expression of EphA2 and Ephrin A-1 in Carcinoma of the Urinary Bladder Clin. Cancer Res., January 15, 2006; 12(2): 353 - 360. [Abstract] [Full Text] [PDF]
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M. Parri, F. Buricchi, M. L. Taddei, E. Giannoni, G. Raugei, G. Ramponi, and P. Chiarugi EphrinA1 Repulsive Response Is Regulated by an EphA2 Tyrosine Phosphatase J. Biol. Chem., October 7, 2005; 280(40): 34008 - 34018. [Abstract] [Full Text] [PDF]
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J. Wykosky, D. M. Gibo, C. Stanton, and W. Debinski EphA2 as a Novel Molecular Marker and Target in Glioblastoma Multiforme Mol. Cancer Res., October 1, 2005; 3(10): 541 - 551. [Abstract] [Full Text] [PDF]
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C. N. Landen Jr., A. Chavez-Reyes, C. Bucana, R. Schmandt, M. T. Deavers, G. Lopez-Berestein, and A. K. Sood Therapeutic EphA2 Gene Targeting In vivo Using Neutral Liposomal Small Interfering RNA Delivery Cancer Res., August 1, 2005; 65(15): 6910 - 6918. [Abstract] [Full Text] [PDF]
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 H Iida, M Honda, H F Kawai, T Yamashita, Y Shirota, B-C Wang, H Miao, and S Kaneko Ephrin-A1 expression contributes to the malignant characteristics of {alpha}-fetoprotein producing hepatocellular carcinoma Gut, June 1, 2005; 54(6): 843 - 851. [Abstract] [Full Text] [PDF]
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 H. Xu, W. Tian, J. N. Lindsley, T. T. Oyama, J. M. Capasso, C. J. Rivard, H. T. Cohen, S. M. Bagnasco, S. Anderson, and D. M. Cohen EphA2: expression in the renal medulla and regulation by hypertonicity and urea stress in vitro and in vivo Am J Physiol Renal Physiol, April 1, 2005; 288(4): F855 - F866. [Abstract] [Full Text] [PDF]
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V. J. Yao, M. G. Ozawa, M. Trepel, W. Arap, D. M. McDonald, and R. Pasqualini Targeting Pancreatic Islets with Phage Display Assisted by Laser Pressure Catapult Microdissection Am. J. Pathol., February 1, 2005; 166(2): 625 - 636. [Abstract] [Full Text] [PDF]
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T. Zhu, B. Starling-Emerald, X. Zhang, K.-O. Lee, P. D. Gluckman, H. C. Mertani, and P. E. Lobie Oncogenic Transformation of Human Mammary Epithelial Cells by Autocrine Human Growth Hormone Cancer Res., January 1, 2005; 65(1): 317 - 324. [Abstract] [Full Text] [PDF]
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C. J. Herrem, T. Tatsumi, K. S. Olson, K. Shirai, J. H. Finke, R. M. Bukowski, M. Zhou, A. L. Richmond, I. Derweesh, M. S. Kinch, et al. Expression of EphA2 is Prognostic of Disease-Free Interval and Overall Survival in Surgically Treated Patients with Renal Cell Carcinoma Clin. Cancer Res., January 1, 2005; 11(1): 226 - 231. [Abstract] [Full Text] [PDF]
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Z.-l. Yuan, Y.-j. Guan, L. Wang, W. Wei, A. B. Kane, and Y. E. Chin Central Role of the Threonine Residue within the p+1 Loop of Receptor Tyrosine Kinase in STAT3 Constitutive Phosphorylation in Metastatic Cancer Cells Mol. Cell. Biol., November 1, 2004; 24(21): 9390 - 9400. [Abstract] [Full Text] [PDF]
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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]
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R. van Doorn, R. Dijkman, M. H. Vermeer, J. J. Out-Luiting, E. M. H. van der Raaij-Helmer, R. Willemze, and C. P. Tensen Aberrant Expression of the Tyrosine Kinase Receptor EphA4 and the Transcription Factor Twist in Sezary Syndrome Identified by Gene Expression Analysis Cancer Res., August 15, 2004; 64(16): 5578 - 5586. [Abstract] [Full Text] [PDF]
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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]
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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]
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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]
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R. L. Pratt and M. S. Kinch Ligand Binding Up-Regulates EphA2 Messenger RNA Through the Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Pathway Mol. Cancer Res., December 1, 2003; 1(14): 1070 - 1076. [Abstract] [Full Text] [PDF]
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G. Zeng, Z. Hu, M. S. Kinch, C.-X. Pan, D. A. Flockhart, C. Kao, T. A. Gardner, S. Zhang, L. Li, L. A. Baldridge, et al. High-Level Expression of EphA2 Receptor Tyrosine Kinase in Prostatic Intraepithelial Neoplasia Am. J. Pathol., December 1, 2003; 163(6): 2271 - 2276. [Abstract] [Full Text]
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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]
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Y.-R. Kao, J.-Y. Shih, W.-C. Wen, Y.-P. Ko, B.-M. Chen, Y.-L. Chan, Y.-W. Chu, P.-C. Yang, C.-W. Wu, and S. R. Roffler Tumor-associated Antigen L6 and the Invasion of Human Lung Cancer Cells Clin. Cancer Res., July 1, 2003; 9(7): 2807 - 2816. [Abstract] [Full Text] [PDF]
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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]
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 A. Palmer and R. Klein Multiple roles of ephrins in morphogenesis, neuronal networking, and brain function Genes & Dev., June 15, 2003; 17(12): 1429 - 1450. [Full Text] [PDF]
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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]
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X. Zhang, T. Zhu, Y. Chen, H. C. Mertani, K.-O. Lee, and P. E. Lobie Human Growth Hormone-regulated HOXA1 Is a Human Mammary Epithelial Oncogene J. Biol. Chem., February 21, 2003; 278(9): 7580 - 7590. [Abstract] [Full Text] [PDF]
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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]
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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]
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J. Walker-Daniels, D. J. Riese II, and M. S. Kinch c-Cbl-Dependent EphA2 Protein Degradation Is Induced by Ligand Binding Mol. Cancer Res., November 1, 2002; 1(1): 79 - 87. [Abstract] [Full Text] [PDF]
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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]
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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]
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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]
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 E. Malaga-Trillo and A. Meyer Genome Duplications and Accelerated Evolution of Hox Genes and Cluster Architecture in Teleost Fishes Integr. Comp. Biol., June 1, 2001; 41(3): 676 - 686. [Abstract] [Full Text] [PDF]
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2 analyses.