Figure 2. EphrinA5 isoforms suppress cell proliferation and migration.

(A) Cells were transfected with 1 µg of pIRESneo-ephrinA5 isoforms or pIRESneo vector, and analyzed at the indicated times by ACP assay. Ectopic expression of ephrinA5 significantly inhibited cell growth in both HepG2 and Hep3B cells as compared to the vector control at 72 hrs. The level of significance was set at p<0.05 (*), p<0.01 (**), or p<0.001 (***). EphrinA5S exerted a stronger suppressive effect than ephrinA5 on both cell lines HepG2 and Hep3B. (B) Cells treated with 3 concentrations of ephrinA5-Fc were analyzed at the indicated time points by MTT assay. EphrinA5-Fc significantly reduced cell proliferation of HepG2 and Hep3B (p<0.05). (C) Expression patterns of Eph receptors in hepatoma cell lines. Primary HCCs and paratumoral tissues were analyzed by RT-PCR. EphB2, A2 and A3 expressed in both cell lines and all human HCC tissues analyzed. (D) Cell migration was compared between HepG2 and Hep3B cells transfected with pIRESneo-ephrinA5 isoforms and vector control. 5 × 10⁴ cells were plated in Transwell inserts and cultured for 24 hr in triplicates. Data were analyzed with Student’s t-tests. Ectopic expression of ephrinA5 significantly decreased the cell migratory ability of both cell types. The level of significance was set at p<0.05 (*), p<0.01 (**), or p<0.001 (***). EphrinA5S also had a stronger inhibotory effect on cell migration.