Figure 2.

Targeting of AKT3 and WEE1 inhibited melanoma tumor growth. Targeting AKT3 and WEE1 synergistically inhibited melanoma tumor growth in vivo. UACC 903 (2A) and 1205 Lu (2B) cells were nucleofected with AKT3 and WEE1 siRNA alone or in combination and 48 hours later, viable cells were s.c. injected into left and right flanks of nude mice. Cells transfected with scrambled siRNA were used as a control. Developing tumors were measured on alternate days for 3 weeks. Significance was measured by one-way analysis of variance, followed by the post hoc test, *P < 0.05, **P < 0.01, ***P < 0.01. Error bars show SEM. Data were obtained from duplicate experiments with 3 mice per group, containing 2 tumors per mouse. C. Western blot analysis showing knockdown of AKT3 and WEE1 in tumor lysates of UACC 903 xenografts harvested at day 9. ERK2 served as a control for equal protein loading. (D) and E, Analysis of size and time matched tumors from animals injected with UACC 903 melanoma cells transfected with scrambled siRNA controls or siRNA to AKT3, WEE1 or AKT3, and WEE1. Tumor sections were immunostained for Ki-67 (2D) or TUNEL (2E) to measure the proliferation and apoptosis, respectively. Bar graphs show the fold change in Ki-67 or TUNEL-positive cells compared with the scrambled siRNA control. Data were obtained from 3 to 4 tumors with 4 to 5 fields averaged per tumor. Significance was measured by one-way analysis of variance, followed by the post hoc test, **P < 0.01, ***P < 0.001, NS; Non-significant. Columns, mean; error bars, SEM.