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Cancer Cell International
Published in: Cancer Cell International 1/2010

Open Access 01-12-2010 | Primary research

Soluble ephrin a1 is necessary for the growth of HeLa and SK-BR3 cells

Authors: Spencer Alford, Adam Watson-Hurthig, Nadia Scott, Amanda Carette, Heather Lorimer, Jessa Bazowski, Perry L Howard

Published in: Cancer Cell International | Issue 1/2010

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Abstract

Background

Ephrin A1 (EFNA1) is a member of the A-type ephrin family of cell surface proteins that function as ligands for the A-type Eph receptor tyrosine kinase family. In malignancy, the precise role of EFNA1 and its preferred receptor, EPHA2, is controversial. Several studies have found that EFNA1 may suppress EPHA2-mediated oncogenesis, or enhance it, depending on cell type and context. However, little is known about the conditions that influence whether EFNA1 promotes or suppresses tumorigenicity. EFNA1 exists in a soluble form as well as a glycophosphatidylinositol (GPI) membrane attached form. We investigated whether the contradictory roles of EFNA1 in malignancy might in part be related to the existence of both soluble and membrane attached forms of EFNA1 and potential differences in the manner in which they interact with EPHA2.

Results

Using a RNAi strategy to reduce the expression of endogenous EFNA1 and EPHA2, we found that both EFNA1 and EPHA2 are required for growth of HeLa and SK-BR3 cells. The growth defects could be rescued by conditioned media from cells overexpressing soluble EFNA1. Interestingly, we found that overexpression of the membrane attached form of EFNA1 suppresses growth of HeLa cells in 3D but not 2D. Knockdown of endogenous EFNA1, or overexpression of full-length EFNA1, resulted in relocalization of EPHA2 from the cell surface to sites of cell-cell contact. Overexpression of soluble EFNA1 however resulted in more EPHA2 distributed on the cell surface, away from cell-cell contacts, and promoted the growth of HeLa cells.

Conclusions

We conclude that soluble EFNA1 is necessary for the transformation of HeLa and SK-BR3 cells and participates in the relocalization of EPHA2 away from sites of cell-cell contact during transformation.
Appendix
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Literature
1.
Gale NW, Holland SJ, Valenzuela DM, Flenniken A, Pan L, Ryan TE, Henkemeyer M, Strebhardt K, Hirai H, Wilkinson DG: Eph receptors and ligands comprise two major specificity subclasses and are reciprocally compartmentalized during embryogenesis. Neuron. 1996, 17: 9-19. 10.1016/S0896-6273(00)80276-7.CrossRefPubMed
2.
Himanen JP, Chumley MJ, Lackmann M, Li C, Barton WA, Jeffrey PD, Vearing C, Geleick D, Feldheim DA, Boyd AW: Repelling class discrimination: ephrin-A5 binds to and activates EphB2 receptor signaling. Nat Neurosci. 2004, 7: 501-509. 10.1038/nn1237.CrossRefPubMed
3.
Wykosky J, Debinski W: The EphA2 receptor and ephrinA1 ligand in solid tumors: function and therapeutic targeting. Mol Cancer Res. 2008, 6: 1795-1806. 10.1158/1541-7786.MCR-08-0244.PubMedCentralCrossRefPubMed
4.
Zelinski DP, Zantek ND, Stewart JC, Irizarry AR, Kinch MS: EphA2 overexpression causes tumorigenesis of mammary epithelial cells. Cancer Res. 2001, 61: 2301-2306.PubMed
5.
Ireton RC, Chen J: EphA2 receptor tyrosine kinase as a promising target for cancer therapeutics. Curr Cancer Drug Targets. 2005, 5: 149-157. 10.2174/1568009053765780.CrossRefPubMed
6.
Walker-Daniels J, Coffman K, Azimi M, Rhim JS, Bostwick DG, Snyder P, Kerns BJ, Waters DJ, Kinch MS: Overexpression of the EphA2 tyrosine kinase in prostate cancer. Prostate. 1999, 41: 275-280. 10.1002/(SICI)1097-0045(19991201)41:4<275::AID-PROS8>3.0.CO;2-T.CrossRefPubMed
7.
Easty DJ, Guthrie BA, Maung K, Farr CJ, Lindberg RA, Toso RJ, Herlyn M, Bennett DC: Protein B61 as a new growth factor: expression of B61 and up-regulation of its receptor epithelial cell kinase during melanoma progression. Cancer Res. 1995, 55: 2528-2532.PubMed
8.
Kinch MS, Carles-Kinch K: Overexpression and functional alterations of the EphA2 tyrosine kinase in cancer. Clin Exp Metastasis. 2003, 20: 59-68. 10.1023/A:1022546620495.CrossRefPubMed
9.
Kinch MS, Moore MB, Harpole DH: Predictive value of the EphA2 receptor tyrosine kinase in lung cancer recurrence and survival. Clin Cancer Res. 2003, 9: 613-618.PubMed
10.
Lu C, Shahzad MM, Wang H, Landen CN, Kim SW, Allen J, Nick AM, Jennings N, Kinch MS, Bar-Eli M: EphA2 overexpression promotes ovarian cancer growth. Cancer Biol Ther. 2008, 7: 1098-1103.PubMedCentralCrossRefPubMed
11.
Sulman EP, Tang XX, Allen C, Biegel JA, Pleasure DE, Brodeur GM, Ikegaki N: ECK, a human EPH-related gene, maps to 1p36.1, a common region of alteration in human cancers. Genomics. 1997, 40: 371-374. 10.1006/geno.1996.4569.CrossRefPubMed
12.
Rosenberg IM, Goke M, Kanai M, Reinecker HC, Podolsky DK: Epithelial cell kinase-B61: an autocrine loop modulating intestinal epithelial migration and barrier function. Am J Physiol. 1997, 273: G824-G832.PubMed
13.
Brantley-Sieders DM, Zhuang G, Hicks D, Fang WB, Hwang Y, Cates JM, Coffman K, Jackson D, Bruckheimer E, Muraoka-Cook RS: The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signaling. J Clin Invest. 2008, 118: 64-78. 10.1172/JCI33154.PubMedCentralCrossRefPubMed
14.
Brantley-Sieders DM, Fang WB, Hicks DJ, Zhuang G, Shyr Y, Chen J: Impaired tumor microenvironment in EphA2-deficient mice inhibits tumor angiogenesis and metastatic progression. FASEB J. 2005, 19: 1884-1886.PubMed
15.
Guo H, Miao H, Gerber L, Singh J, Denning MF, Gilliam AC, Wang B: Disruption of EphA2 receptor tyrosine kinase leads to increased susceptibility to carcinogenesis in mouse skin. Cancer Res. 2006, 66: 7050-7058. 10.1158/0008-5472.CAN-06-0004.CrossRefPubMed
16.
Zantek ND, Azimi M, Fedor-Chaiken M, Wang B, Brackenbury R, Kinch MS: E-cadherin regulates the function of the EphA2 receptor tyrosine kinase. Cell Growth Differ. 1999, 10: 629-638.PubMed
17.
Fang WB, Ireton RC, Zhuang G, Takahashi T, Reynolds A, Chen J: Overexpression of EPHA2 receptor destabilizes adherens junctions via a RhoA-dependent mechanism. J Cell Sci. 2008, 121: 358-368. 10.1242/jcs.017145.CrossRefPubMed
18.
Hess AR, Seftor EA, Gruman LM, Kinch MS, Seftor RE, Hendrix MJ: VE-cadherin regulates EphA2 in aggressive melanoma cells through a novel signaling pathway: implications for vasculogenic mimicry. Cancer Biol Ther. 2006, 5: 228-233. 10.4161/cbt.5.2.2510.CrossRefPubMed
19.
Miura K, Nam JM, Kojima C, Mochizuki N, Sabe H: EphA2 engages Git1 to suppress Arf6 activity modulating epithelial cell-cell contacts. Mol Biol Cell. 2009, 20: 1949-1959. 10.1091/mbc.E08-06-0549.PubMedCentralCrossRefPubMed
20.
Cui XD, Lee MJ, Yu GR, Kim IH, Yu HC, Song EY, Kim DG: EFNA1 ligand and its receptor EphA2: Potential biomarkers for hepatocellular carcinoma. Int J Cancer. 2010, 126 (4): 940-9.PubMed
21.
Bartley TD, Hunt RW, Welcher AA, Boyle WJ, Parker VP, Lindberg RA, Lu HS, Colombero AM, Elliott RL, Guthrie BA: B61 is a ligand for the ECK receptor protein-tyrosine kinase. Nature. 1994, 368: 558-560. 10.1038/368558a0.CrossRefPubMed
22.
Easty DJ, Hill SP, Hsu MY, Fallowfield ME, Florenes VA, Herlyn M, Bennett DC: Up-regulation of ephrin-A1 during melanoma progression. Int J Cancer. 1999, 84: 494-501. 10.1002/(SICI)1097-0215(19991022)84:5<494::AID-IJC8>3.0.CO;2-O.CrossRefPubMed
23.
Potla L, Boghaert ER, Armellino D, Frost P, Damle NK: Reduced expression of EphrinA1 (EFNA1) inhibits three-dimensional growth of HT29 colon carcinoma cells. Cancer Lett. 2002, 175: 187-195. 10.1016/S0304-3835(01)00613-9.CrossRefPubMed
24.
Abraham S, Knapp DW, Cheng L, Snyder PW, Mittal SK, Bangari DS, Kinch M, Wu L, Dhariwal J, Mohammed SI: Expression of EphA2 and Ephrin A-1 in carcinoma of the urinary bladder. Clin Cancer Res. 2006, 12: 353-360. 10.1158/1078-0432.CCR-05-1505.CrossRefPubMed
25.
Herath NI, Spanevello MD, Sabesan S, Newton T, Cummings M, Duffy S, Lincoln D, Boyle G, Parsons PG, Boyd AW: Over-expression of Eph and ephrin genes in advanced ovarian cancer: ephrin gene expression correlates with shortened survival. BMC Cancer. 2006, 6 (144):
26.
Wykosky J, Gibo DM, Stanton C, Debinski W: EphA2 as a novel molecular marker and target in glioblastoma multiforme. Mol Cancer Res. 2005, 3: 541-551. 10.1158/1541-7786.MCR-05-0056.CrossRefPubMed
27.
Liu DP, Wang Y, Koeffler HP, Xie D: Ephrin-A1 is a negative regulator in glioma through down-regulation of EphA2 and FAK. Int J Oncol. 2007, 30: 865-871.PubMed
28.
Nakamura R, Kataoka H, Sato N, Kanamori M, Ihara M, Igarashi H, Ravshanov S, Wang YJ, Li ZY, Shimamura T: EPHA2/EFNA1 expression in human gastric cancer. Cancer Sci. 2005, 96: 42-47. 10.1111/j.1349-7006.2005.00007.x.CrossRefPubMed
29.
Noblitt LW, Bangari DS, Shukla S, Knapp DW, Mohammed S, Kinch MS, Mittal SK: Decreased tumorigenic potential of EphA2-overexpressing breast cancer cells following treatment with adenoviral vectors that express EphrinA1. Cancer Gene Ther. 2004, 11: 757-766. 10.1038/sj.cgt.7700761.CrossRefPubMed
30.
Shi L, Itoh F, Itoh S, Takahashi S, Yamamoto M, Kato M: Ephrin-A1 promotes the malignant progression of intestinal tumors in Apc(min/+) mice. Oncogene. 2008, 27 (23): 3265-73. 10.1038/sj.onc.1210992.CrossRefPubMed
31.
Wykosky J, Palma E, Gibo DM, Ringler S, Turner CP, Debinski W: Soluble monomeric EphrinA1 is released from tumor cells and is a functional ligand for the EphA2 receptor. Oncogene. 2008, 27: 7260-7273. 10.1038/onc.2008.328.PubMedCentralCrossRefPubMed
32.
Pandey A, Shao H, Marks RM, Polverini PJ, Dixit VM: Role of B61, the ligand for the Eck receptor tyrosine kinase in TNF-alpha-induced angiogenesis. Science. 1995, 268: 567-569. 10.1126/science.7536959.CrossRefPubMed
33.
Holzman LB, Marks RM, Dixit VM: A novel immediate-early response gene of endothelium is induced by cytokines and encodes a secreted protein. Mol Cell Biol. 1990, 10: 5830-5838.PubMedCentralCrossRefPubMed
34.
Davis S, Gale NW, Aldrich TH, Maisonpierre PC, Lhotak V, Pawson T, Goldfarb M, Yancopoulos GD: Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science. 1994, 266: 816-819. 10.1126/science.7973638.CrossRefPubMed
35.
Alford SC, Bazowski J, Lorimer H, Elowe S, Howard PL: Tissue transglutaminase clusters soluble A-type ephrins into functionally active high molecular weight oligomers. Exp Cell Res. 2007, 313: 4170-4179. 10.1016/j.yexcr.2007.07.019.CrossRefPubMed
36.
Peehl DM, Stanbridge EJ: Anchorage-independent growth of normal human fibroblasts. Proc Natl Acad Sci USA. 1981, 78: 3053-3057. 10.1073/pnas.78.5.3053.PubMedCentralCrossRefPubMed
37.
Orsulic S, Kemler R: Expression of Eph receptors and ephrins is differentially regulated by E-cadherin. J Cell Sci. 2000, 113 (Pt 10): 1793-1802.PubMed
38.
Holland SJ, Gale NW, Gish GD, Roth RA, Songyang Z, Cantley LC, Henkemeyer M, Yancopoulos GD, Pawson T: Juxtamembrane tyrosine residues couple the Eph family receptor EphB2/Nuk to specific SH2 domain proteins in neuronal cells. EMBO J. 1997, 16: 3877-3888. 10.1093/emboj/16.13.3877.PubMedCentralCrossRefPubMed
39.
Holland SJ, Gale NW, Mbamalu G, Yancopoulos GD, Henkemeyer M, Pawson T: Bidirectional signalling through the EPH-family receptor Nuk and its transmembrane ligands. Nature. 1996, 383: 722-725. 10.1038/383722a0.CrossRefPubMed
40.
Dalva MB, Takasu MA, Lin MZ, Shamah SM, Hu L, Gale NW, Greenberg ME: EphB receptors interact with NMDA receptors and regulate excitatory synapse formation. Cell. 2000, 103: 945-956. 10.1016/S0092-8674(00)00197-5.CrossRefPubMed
Metadata
Title
Soluble ephrin a1 is necessary for the growth of HeLa and SK-BR3 cells
Authors
Spencer Alford
Adam Watson-Hurthig
Nadia Scott
Amanda Carette
Heather Lorimer
Jessa Bazowski
Perry L Howard
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2010
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/1475-2867-10-41

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