Fig. 7

PHB2-mediated SHIP2 degradation leads to Akt activation and GC proliferation. A Knockdown of PHB2 increased SHIP2 protein expression and diminished Akt activation, which was reversed by co-knockdown of SHIP2. Data shown represent three independent experiments. B, C Knockdown of PHB2 inhibited GC cell proliferation, which was abolished by co-knockdown of SHIP2 as shown in clonogenic assays (B) and BrdU incorporation (C). Data are representatives or mean ± SEM; n = 3 independent experiments, one-way ANOVA followed by Tukey’s multiple comparison. Scale bar, 1 cm. D-F, Overexpression of myr-Akt reversed the inhibition of Akt signaling (D) and GC cell proliferation caused by silencing of PHB2 as shown in clonogenic assays (E) and BrdU incorporation (F). Data are representatives or mean ± SEM; n = 3 independent experiments, two-tailed Student’s t-test. Scale bar, 1 cm. G-I Representative Photographs (G), tumour weight (H) and growth curves (I) of MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts in nu/nu mice with or without co-transduction of myr-Akt. Data are representatives or mean ± SEM; n = 3 mice per group, one-way ANOVA followed by Tukey’s multiple comparison (H), two-way ANOVA followed by Tukey’s multiple comparison (I). J Western blotting showing the expression of PHB2, SHIP2 and Akt in GC cells isolated from MKN-28.sh-scramble and MKN-28.sh-PHB2 xenografts harvested from mice treated as described in G. Data are representatives of three independent experiments