Effect of ephrin-A1/EphA2 on invasion of trophoblastic cells

Abstract

The effect of axon guidance factors ephrin-A1/EphA2 on the invasion of trophoblastic cells and the possible mechanism were investigated in this study. The expression of EphA2 in vascular endothelial cells was detected by immunohistochemistry. The proliferation and invasion of TEV-1 cells (an extravillous trophoblastic cell line) in first trimester were determined by cell counting kit-8 (CCK-8) and Transwell invasion assay. Real-time PCR was used to detect the expression of ephrin-A1 in TEV-1 cells treated with EphA2 at different concentrations (10, 50, 100, 500, 1000 and 5000 μg/L). The results showed: (1) EphA2 was expressed in the vascular endothelial cells; (2) EphA2 could promote the proliferation of TEV-1 cells. The proliferative capacity reached a peak in TEV-1 cells treated with 100 μg/L EphA2 (P<0.05); (3) EphA2 could increase the invasion of TEV-1 cells. The invasive ability was the greatest in TEV-1 cells treated with 500 μg/L EphA2 (P<0.05); (4) in the presence of EphA2 (0–500 μg/L), the expression of ephrin-A1 was increased concentration-dependently (P<0.05), but when the concentration of EphA2 was over 500 μg/L, the expression of ephrin-A1 ceased to increase (P>0.05). It was concluded that EphA2 can promote the invasion and proliferation of the human extravillous trophoblastic cells probably by regulating the ephrin-A1 ligand.

Keywords

axon guidance factor / receptors / extravillous trophoblastic cells

References

[1] ReynoldsL.P., BorowiczP.P., VonnahmeK.A., et al.. Animal models of placental angiogenesis. Placenta, 2005, 26(10): 689-708

[2] SawamiphakS., SeidelS., EssmannC.L., et al.. Ephrin-B2 regulates VEGFR2 function in developmental and tumour angiogenesis. Nature, 2010, 465(7297): 487-491

[3] Goldman-WohlD., GreenfieldC., Haimov-KochmanI., et al.. Eph and ephrin expression in normal placental development and preeclampsia. Placenta, 2004, 25(7): 623-630

[4] FengH.C., ChoyM.Y., DengW., et al.. Establishment and characterization of a human first-trimester extravillous trophoblast cell line (TEV-1). J Soc Gyneco Investig, 2005, 12(4): 21-32

[5] JaffeE.A., HoyerL.W., NachmanR.L.. Synthesis of von Willebrand factor by cultured human endothelial cells. Proc Natl Acad Sci, 1974, 71(5): 1906-9190

[6] RaffanielloR., FedorovaD., LpD., et al.. Hsp90 co-localizes with Rav-GDl-1 and regulates agonist-induced amylase release in AR22J cells. Cell Physiol Biochem, 2009, 24(5–6): 369-378

[7] LivakK.J., SchmittgenT.D.. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C (T)) Method. Methods, 2001, 25(4): 402-408

[8] GuibourdencheJ., FournierT., MalassineA., et al.. Development and hormonal functions of the human placenta. Folia Histoche Cytobiol, 2009, 47(5): S35-S40

[9] HarrisL.K.. Review: Trophoblast-vascular cell interactions in first trimester: how to remodel a vessel. Placenta, 2010, 31(suppl): S93-S98

[10] ReberM., HindgesR., LemkeG.. Eph receptors and ephrin ligands in axon guidance. Adv Exp Med Biol, 2007, 621: 32-49

[11] PasqualeE.B.. Eph receptors and ephrins in cancer: bidiretional signaling and beyond. Nat Rev Cancer, 2010, 10(3): 165-180