Figure 6. Twist1 is required for AKT1-mediated EMT repression.
(A) MCF 10A cells were continuously treated with or without TGFβ or MK-2206. Phenotypic changes were examined by Western blot with the indicated antibodies.
(B) MCF 10A cells harboring shCTRL and shTwist1 were treated with 0.2 μM or 0.5 μM MK-2206 for 10 days and subjected to Western blotting with the indicated antibodies.
(C) Resveratrol reduces MK-2206-induced EMT. MCF 10A cells were continuously treated with or without MK-2206 or Resveratrol and subjected to Western blotting with the indicated antibodies.
(D and E) The number of migrating cells and their distance of migration (arbitrary unit) calculated from three independent views.
(F) Representative photograph of metastatic lung nodules. 4T1 cells (1 × 105 cells/0.1 ml) were injected into the lateral tail vein of Balb/c female mice. MK-2206 and resveratrol were then administered orally at a dosage of 200 mg/kg (n = 6/group) per day for 21 days. Mice were sacrificed, and the lungs were stained with India ink and resected for imaging.
(G) Quantification of the lung nodules. Error bars represent ± SD for n = 6.
(H) The survival time of mice in (F). Survival curve was plotted by Kaplan-Meier analysis.
(I) Schematic diagram showing the combinatorial strategy of utilizing resveratrol to overcome MK-2206-induced metastatic potential.