Fig. 6

AKT is a therapeutic target in the CRC cells. a Effect of the knockdown of AKT1/2 on total AKT protein. The CRC cells stably expressing AKT1/2 knockdown or control were established. The expression was determined by western blotting. b The cell growth in the CRC cells after AKT1/2 knockdown by the CCK-8 assay at different time points. c The effect of CTD treatment on the anchorage-independent growth was assessed in the CRC cells expressing pLKO or shRNA-AKT1/2. The anchorage-independent cell growth is decreased with or without CTD treatment after AKT1/2 expression was decreased. d The ability of migration was detected by the transwell assay after the knockdown of the AKT1/2. e The ability of invasiveness was detected by the transwell assay after the knockdown of the AKT1/2. f The AKT expression level of pLKO, shRNA-AKT1/2, overexpressed pUSE-CA-Akt1/2 and mock by western blot analyzed. g and h The proliferation of cells expressing pLKO or shRNA-AKT1/2 or overexpressing pUSE-CA-Akt1/2 and corresponding mock control were examined by the MTT and anchorage-independent cell growth assays. All the data are shown as means ± SD of values from triplicate samples. (*p < 0.05, **p < 0.01, *** p < 0.001) indicate a significant difference compared to the control. i The additive effects were observed via the coordinated inhibition of MDM2-p53 interaction and AKT, as shown by immunoblotting. MDM2, p53, and AKT were evaluated by immunoblotting after treatment with CTD for 24 h and infection with lentiviral AKT shRNA for 24 h. Actin is used as a loading control