The PI3K pathway was involved in DIXDC1‐induced p21, cyclin D1 alteration. (a) DLD1‐DIXDC1‐1 cells were treated with 10 µM LY294002 or DMSO vehicle control for 12 h and 24 h, then were harvested and subjected to immunoblotting analysis using Akt, p‐Akt, p21, cyclin D1, and β‐catenin antibodies. (b) Blocking the PI3K pathway with LY294002 decreased the S‐phase cell fraction in DLD1‐DIXDC1‐1 cells. Cells were treated with LY292002 (10 µM) or DMSO for 12 h or 24 h, and were then collected for flow cytometry analysis. The percentages of cells in the G0/G1, S, and G2/M phases were counted. Columns represent the mean of three independent experiments; bars, SD. *P < 0.01, compared with DMSO 12 h; **P < 0.01, compared with DMSO 24 h. (c) DIXDC1 interacted with p‐Akt in DIXDC1 overexpression cells. Cell lysates were immunoprecipitated with anti‐DIXDC1 antibody. The immunoprecipitates and cell lysates were probed with anti‐phospho‐Akt, anti‐Akt, and anti‐DIXDC1 antibodies. IP, immnunoprecipitation; IB, immunoblot.