1. RAS/RAF pathway-activated ovarian cancer cells exhibit MEK dependence and synergistic induction of apoptosis with combined MEK/AKT inhibition.
RAS/RAF pathway-activated ovarian cancer cells exhibit MEK dependence and synergistic induction of apoptosis with combined MEK/AKT inhibition. A, levels of activated RAS (RAS-GTP) were determined by pulldown of GTP-bound RAS using recombinant RAS binding domain of RAF. Input lysates were also immunoblotted as shown. B, IC50 values for the allosteric inhibitor of MEK1/2, PD (PD901) were calculated after treatment with 0 to 500 nM of inhibitor for 5 days. Immunoblots for p-ERK and ERK after treatment with PD901 (50 nM) for 0 to 24 hours. C, induction of cell death after combined treatment with 50 nM PD901 and increasing doses of either AKTi1/2 or MK2206 (2 or 10 μM) was measured by FACS analysis at 72 hours in OVCAR-5 cells. Maximal cell death was induced after cotreatment with PD901 and MK2206 (asterisk indicates P ≤ versus all other treatments, n ≥ 3). D, OVCAR-5 cells were treated with PD901 (50 nM) or MK2206 (10 μM) alone or in combination for 48 hours and lysates were immunoblotted.
Aphrothiti J. Hanrahan et al. Cancer Discov 2012;2:56-67
©2012 by American Association for Cancer Research