Fig. 6.

Effect of VNLG-152 on f-AR activity and eIF4F complex formation in 22Rv1 cells in vitro. (A) Effects of VNLG-152 on DHT-induced increase in f-AR reporter activity in 22Rv1/f-AR cells. 22Rv1/f-AR cells were seeded at a density of 0.1 × 10⁵ cells·mL⁻¹ of medium for 24 h. Then the medium was changed to RPMI 1640 without fetal bovine serum, and the cells were treated with vehicle (control) or DHT (10 nm) alone or in combination with VNLG-152 (5 and 10 μM) for 24 h. Luciferase activity was measured to etermine androgen receptor reporter activity in the 22Rv1/f-AR cells. Values are presented as the mean ± SEM from three separate experiments. *P < 0.01 compared with DHT-treated cells. Statistical significance was determined by Student’s t test and ANOVA. (B) Effect of VNLG-152 on eIF4E and eIF4G interaction in 22Rv1 cells. Western blot analysis of 4E-BP1, eIF4G and eIF4E from m⁷GTP purifications of vehicle- and VNLG-152 (5 μM, 24 h)-treated cell lysates. (C) 22Rv1 cells were treated with VNLG-152 (5 μM, 24 h) to evaluate the effect of p-mTOR and p-4E-BP1 by western blotting. Equal protein concentrations of cell lysate were separated by SDS/PAGE and western blots probed with indicated antibodies.