To confirm whether blocking the PI3K/Akt pathway inhibits cell proliferation and invasiveness and promotes apoptosis of YTS cells, LY-294002, a specific inhibitor of PI3K, which can significantly inhibit the protein expression of p-Akt and PI3K, but not Akt, was used to treat cells (Fig. 4A,B). As expected, CCK-8 and colony formation assays showed that blocking the PI3K-Akt pathway caused increased cell proliferation and colony formation of YTS cells (Fig. 4C,D). Flow cytometry analysis and transwell invasion assay exhibited that blocking the PI3K/Akt pathway resulted in increased apoptosis and diminished cell invasion ability in YTS cells (Fig. 4E,F). Fig. 4.Blocking the PI3K/AKT pathway inhibits YTS cell proliferation and invasion and promotes apoptosis. (A,B) Western blot analysis revealed that LY294002 treatment (5 μM and 20 μM) significantly suppressed p-AKT and PI3K in a dose-dependent manner, while AKT expression in the three experimental groups was not significantly different. (C) CCK-8 assay showed that LY294002 treatment had an inhibitory effect on cell proliferation of YTS cells in a dose-dependent manner at 48 h. (D) Colony formation assay showed that LY294002 treatment caused a decrease in the clonogenic survival of YTS cells in a dose-dependent manner. (E) Flow cytometric analysis exhibited that LY294002 treatment had a promotive effect on proliferation of YTS cells in a dose-dependent manner at 48 h. (F) Transwell invasion assay showed that LY294002 treatment had an inhibitory effect on the cell invasion ability of YTS-GSN cells in a dose-dependent manner at 48 h. *P<0.05,**P<0.01.