Molecular and Cellular Biochemistry

, Volume 361, Issue 1–2, pp 259–265 | Cite as

Activation of erythropoietin-producing hepatocellular receptor A2 attenuates cell adhesion of human fallopian tube epithelial cells via focal adhesion kinase dephosphorylation



Tyrosine kinase receptor erythropoietin-producing hepatocellular receptor A2 (EphA2) and its predominant ligand EphrinA1 have been studied extensively for their roles of mediating cell adhesion in epithelial cells. However, EphA2 signaling in human fallopian tube epithelial cells is poorly understood. In this study, primary cultured fallopian tube epithelial cells were used as a model treated with EphrinA1-Fc or IgG-Fc (control), to explore the role of EphA2 signal and its network involved in the regulation of cell adhesion of tubal epithelia cells. The activation of EphA2 and focal adhesion kinase (FAK) was evaluated by western blotting assay in the cultured fallopian tube epithelia cells, of which the cell adhesion activity was determined by MTT assay. A significantly negative correlation was found between phosphorylated-EphA2 (Pho-EphA2) and phosphorylated-FAK (Pho-FAK) after exposure to EphrinA1-Fc (P = 0.000; r = −0.848). EphrinA1-Fc increased Pho-EphA2 and reduced Pho-FAK in seconds, with the apex level of Pho-EphA2 and the nadir level of Pho-FAK detected at the same time (10 min). Cell adhesion of the cultured cells supplemented with EphrinA1-Fc appeared to be weaker than that of the controls at the later time points of the treatment (from 30 to 120 min) (P < 0.05). Taken together, the EphrinA1 addition directly induces an elevated Pho-EphA2 accompanied by a decreased Pho-FAK in human fallopian tube epithelia cells. Furthermore, activation of EphA2 participates in the regulation of fallopian tube cell adhesion via FAK dephosphorylation.


Erythropoietin-producing hepatocellular receptor A2 EphrinA1 Focal adhesion kinase Fallopian tube Cell adhesion 


  1. 1.
    Davy A, Aubin J, Soriano P (2004) Ephrin-B1 forward and reverse signaling are required during mouse development. Genes Dev 18:572–583PubMedCrossRefGoogle Scholar
  2. 2.
    Carter N, Nakamoto T, Hirai H, Hunter T (2002) EphrinA1-induced cytoskeletal re-organization requires FAK and p130(cas). Nat Cell Biol 4:565–573PubMedGoogle Scholar
  3. 3.
    Lim BK, Matsuda N, Poo MM (2008) Ephrin-B reverse signaling promotes structural and functional synaptic maturation in vivo. Nat Neurosci 11:160–169PubMedCrossRefGoogle Scholar
  4. 4.
    Zhao C, Irie N, Takada Y, Shimoda K, Miyamoto T, Nishiwaki T, Suda T, Matsuo K (2006) Bidirectional ephrinB2-EphB4 signaling controls bone homeostasis. Cell Metab 4:111–121PubMedCrossRefGoogle Scholar
  5. 5.
    Carles-Kinch K, Kilpatrick KE, Stewart JC, Kinch MS (2002) Antibody targeting of the EphA2 tyrosine kinase inhibits malignant cell behavior. Cancer Res 62:2840–2847PubMedGoogle Scholar
  6. 6.
    Zelinski DP, Zantek ND, Stewart JC, Irizarry AR, Kinch MS (2001) EphA2 overexpression causes tumorigenesis of mammary epithelial cells. Cancer Res 61:2301–2306PubMedGoogle Scholar
  7. 7.
    Noblitt LW, Bangari DS, Shukla S, Mohammed S, Mittal SK (2005) Immunocompetent mouse model of breast cancer for preclinical testing of EphA2-targeted therapy. Cancer Gene Ther 12:46–53PubMedCrossRefGoogle Scholar
  8. 8.
    Guan JL, Shalloway D (1992) Regulation of focal adhesion-associated protein tyrosine kinase by both cellular adhesion and oncogenic transformation. Nature 358:690–692PubMedCrossRefGoogle Scholar
  9. 9.
    Chao WT, Kunz J (2009) Focal adhesion disassembly requires clathrin-dependent endocytosis of integrins. FEBS Lett 583:1337–1343PubMedCrossRefGoogle Scholar
  10. 10.
    Askari JA, Tynan CJ, Webb SE, Martin-Fernandez ML, Ballestrem C, Humphries MJ (2010) Focal adhesions are sites of integrin extension. J Cell Biol 188:891–903PubMedCrossRefGoogle Scholar
  11. 11.
    Wang J, Armant DR (2002) Integrin-mediated adhesion and signaling during blastocyst implantation. Cells Tissues Organs 172:190–201PubMedCrossRefGoogle Scholar
  12. 12.
    Wang J, Mayernik L, Armant DR (2007) Trophoblast adhesion of the peri-implantation mouse blastocyst is regulated by integrin signaling that targets phospholipase C. Dev Biol 302:143–153PubMedCrossRefGoogle Scholar
  13. 13.
    Ilic D, Genbacev O, Jin F, Caceres E, Almeida EA, Bellingard-Dubouchaud V, Schaefer EM, Damsky CH, Fisher SJ (2001) Plasma membrane-associated pY397FAK is a marker of cytotrophoblast invasion in vivo and in vitro. Am J Pathol 159:93–108PubMedCrossRefGoogle Scholar
  14. 14.
    Daoud G, Le bellego F, Lafond J (2008) PP2 regulates human trophoblast cells differentiation by activating p38 and ERK1/2 and inhibiting FAK activation. Placenta 29:862–870PubMedCrossRefGoogle Scholar
  15. 15.
    Miao H, Burnett E, Kinch M, Simon E, Wang B (2000) Activation of EphA2 kinase suppresses integrin function and causes focal-adhesion-kinase dephosphorylation. Nat Cell Biol 2:62–69PubMedCrossRefGoogle Scholar
  16. 16.
    Liu DP, Wang Y, Koeffler HP, Xie D (2007) Ephrin-A1 is a negative regulator in glioma through down-regulation of EphA2 and FAK. Int J Oncol 30:865–871PubMedGoogle Scholar
  17. 17.
    Lindberg RA, Hunter T (1990) cDNA cloning and characterization of Eck, an epithelial cell receptor protein-tyrosine kinase in the Eph/Elk family of protein kinases. Mol Cell Biol 10:6316–6324PubMedGoogle Scholar
  18. 18.
    Coffman KT, Hu M, Carles-Kinch K, Tice D, Donacki N, Munyon K, Kifle G, Woods R, Langermann S, Kiener PA, Kinch MS (2003) Differential EphA2 epitope display on normal versus malignant cells. Cancer Res 63:7907–7912PubMedGoogle Scholar
  19. 19.
    Wykosky J, Debinski W (2008) The EphA2 receptor and ephrinA1 ligand in solid tumors: function and therapeutic targeting. Mol Cancer Res 6:1795–1806PubMedCrossRefGoogle Scholar
  20. 20.
    Huveneers S, Truong H, Fassler R, Sonnenberg A, Danen EH (2008) Binding of soluble fibronectin to integrin alpha5 beta1-link to focal adhesion redistribution and contractile shape. J Cell Sci 121:2452–2462PubMedCrossRefGoogle Scholar
  21. 21.
    Fujii H, Tatsumi K, Kosaka K, Yoshioka S, Fujiwara H, Fujii S (2006) Eph-ephrin A system regulates murine blastocyst attachment and spreading. Dev Dyn 235:3250–3258PubMedCrossRefGoogle Scholar
  22. 22.
    Goldman-Wohl D, Greenfield C, Haimov-Kochman R, Ariel I, Anteby EY, Hochner-Celnikier D, Farhat M, Yagel S (2004) Eph and ephrin expression in normal placental development and preeclampsia. Placenta 25:623–630PubMedCrossRefGoogle Scholar
  23. 23.
    Yoshimura Y, Miyakoshi K, Hamatani T, Iwahashi K, Takahashi J, Kobayashi N, Sueoka K, Miyazaki T, Kuji N, Tanaka M (1998) Role of beta 1 integrins in human endometrium and decidua during implantation. Horm Res 50(Suppl 2):46–55PubMedCrossRefGoogle Scholar
  24. 24.
    Shiokawa S, Yoshimura Y, Nagamatsu S, Sawa H, Hanashi H, Sakai K, Noguchi K, Nakamura Y (1998) Functional role of focal adhesion kinase in the process of implantation. Mol Hum Reprod 4:907–914PubMedCrossRefGoogle Scholar
  25. 25.
    Hanashi H, Shiokawa S, Akimoto Y, Sakai K, Suzuki N, Kabir-Salmani M, Nagamatsu S, Iwashita M, Nakamura Y (2003) Physiologic role of decidual beta1 integrin and focal adhesion kinase in embryonic implantation. Endocr J 50:189–198PubMedCrossRefGoogle Scholar
  26. 26.
    Brantley-Sieders DM, Zhuang G, Hicks D, Fang WB, Hwang Y, Cates JM, Coffman K, Jackson D, Bruckheimer E, Muraoka-Cook RS, Chen J (2008) The receptor tyrosine kinase EphA2 promotes mammary adenocarcinoma tumorigenesis and metastatic progression in mice by amplifying ErbB2 signaling. J Clin Investig 118:64–78PubMedCrossRefGoogle Scholar
  27. 27.
    Rodgrigo JP, Alvarez-Alija G, Menendez ST, Mancebo G, Allonca E, Garcia Carracedo D, Fresno M, Suarez-Nieto C, Garcia-Pedrero JM (2011) Cortactin and focal adhesion kinase as predictors of cancer risk in patients with laryngeal premalignancy. Cancer Prev Res 4:1333–1341CrossRefGoogle Scholar
  28. 28.
    Daves MH, Hilsenbeck SG, Lau CC, Man TK (2011) Meta-analysis of multiple microarray datasets reveals a common gene signature of metastasis in solid tumors. BMC Med Genomics 4:56PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  1. 1.Institute of Reproductive Immunology, College of Life Science and TechnologyJinan UniversityGuangzhou CityPeople’s Republic of China

Personalised recommendations