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. 2020 Aug 15;17(3):707–725. doi: 10.20892/j.issn.2095-3941.2020.0056

Figure 7.

Figure 7

Rescued expression of SHP2 in SHP2 deleted cells restores the proliferation ability of breast cancer cells. (A) Rescued expression of SHP2 restores the expression of phosphorylated AKT (T308), phosphorylated ERK (T202/Y204), phosphorylated GSK3β (Ser9), and Cyclin D1 in SHP2 knockout cells. SHP2 knockout cells were infected with lentivirus for SHP2 expression. The cells were then lysed and analyzed via western blotting. (B) CCK-8 assays showed that the re-expression of SHP2 in SHP2 deleted cells restored the cell proliferation ability. Data are shown as mean ± SD. Statistical analysis was performed with two-way ANOVA (****P < 0.0001). (C) Rescued expression of SHP2 restored the colony formation ability of SHP2 knockout cells. Statistical analysis was carried out with one-way ANOVA (****P < 0.0001). (D) Rescued SHP2 expression resulted in a smaller proportion of G1 phase cells than that in SHP2 knockout cells. Data are shown as mean ± SD. Statistical analysis was carried out with one-way ANOVA (****P < 0.0001). (E) EdU incorporation assays showed that the re-expression of SHP2 increased the proportion of cells in S phase. Data are expressed as mean ± SD from 6 independent fields (****P < 0.0001). (F) Rescued SHP2 in SHP2 deleted cells restored the nuclear expression of Cyclin D1. Data are expressed as mean ± SD from 5 independent fields (****P < 0.0001). (G) SHP2 knockout inhibited tumor growth in vivo, whereas the re-expression of SHP2 rescued tumor growth defects caused by SHP2 deletion. Statistical analysis was performed with two-way ANOVA (****P < 0.0001). (H) SHP2 knockout in breast cancer cells resulted in a significant decrease in tumor weight, whereas the rescued expression of SHP2 in SHP2 deleted cells resulted in tumors heavier than those in the control group. All data are expressed as mean ± SD (****P < 0.0001). (I) SHP2 knockout decreased the expression of Ki67 and Cyclin D1 in tumor sections, whereas rescued expression of SHP2 restored the expression of these 2 proteins (**P < 0.01, ***P < 0.001, ****P < 0.0001).