Figure 1.
Cell viability effect of GDC0941 and AZD6244 in BxPC-3, MIA PaCa-2, PANC-1 and Capan-2 pancreatic cancer cell lines.
Cells were treated with varying concentrations of GDC0941 (A) or AZD6244 (B) alone for 72 hours. Doses ranged from 1 nM to 10 µM.
Figure 2.
Synergistic effects of GDC0941-AZD6244 combination therapy on cell viability and proliferation.
BxPC-3 (A) and MIA PaCa-2 cells (B) were treated with GDC0941 alone, AZD6244 alone or GDC0941-AZD6244 in combination. Results were analyzed according to the Chou-Talalay method [18]. The combination index (CI) values were calculated by using CalcuSyn software. PANC-1 (C) and Capan-2 cell line (D) were treated with GDC0941 at 400 nM, AZD6244 at 200 nM or combination. * p value <0.05 compared with control or single agent alone.
Figure 3.
Effects of GDC0941 and AZD6244 therapy on cell apoptosis in pancreatic cancer cell lines.
Pancreatic cancer cell lines were treated with GDC0941 alone, AZD6244 alone or GDC0941-AZD6244 in combination for 72 hours. Cell apoptosis was detected by flow cytometry. A, B: combinations vs. untreated groups (*p values <0.05), combinations vs. single agents (*p values <0.05), single agents vs. untreated groups (*p values <0.05). C, D: combinations vs. untreated groups (*p value <0.05) and combinations vs. single agents (*p value <0.05).
Figure 4.
Effects of GDC0941-AZD6244 combination therapy on PI3K/AKT and MEK/ERK pathways.
All four pancreatic cancer cell lines were treated with the GDC0941-AZD6244 combination for 24 hours, Cell lysates were then harvested to detect p-AKT (S473)/AKT, p-ERK (T202/Y204)/ERK, p-S6 (S240/244)/S6 and p-4E-BP1 (S65)/4E-BP1.
Figure 5.
Effects of GDC0941 and AZD6244 on PI3K/AKT and MEK/ERK pathways of MIA PaCa-2 and PANC-1 cell lines.
Both MIA PaCa-2 and PANC-1 cells were treated with 400 nm GDC0941, 200 nM AZD6244 or a combination at these doses for 24 hours. Cell proteins were then harvested to detect p-AKT(S473)/AKT, p-ERK(T202/Y204)/ERK, p-S6(S240/244)/S6 and p-4E-BP1(S65)/4E-BP-1.
Figure 6.
Effects of GDC0941 and AZD6244 on BxPC-3 xenograft model.
A: 1×106 BxPC-3 pancreatic cancer cells were injected s.c. into the right flank of female nude mice. Mice received vehicle (0.5% methylcellulose /0.2% Tween-20), 50 mg/kg GDC0941 QD, 25 mg/kg AZD6244 BID, or 50 mg/kg GDC0941 QD plus 25 mg/kg AZD6244 BID orally for 18 days. Data are presented as mean ± SE. *p values were determined by unpaired t-test. B: Kaplan-Meier survival curves of xenografts receiving vehicle, GDC0941 alone, AZD6244 alone and GDC0941-AZD6244 in combination.