Fig. 2. Ursolic acid (UA) plus 3,3’-diindolylmethane (DIM) induces G1 phase arrest in ESCC.
(A) UA plus DIM induced G1 phase arrest in TE-8 and TE-12 cells. Cells were treated with UA (20 μM), DIM (50 μM), and a combination of both for 48 h. PI staining-based flow cytometry assay was designed to study the cell cycle in esophageal cancer cells. (B) UA plus DIM combination suppressed G1 phase-related protein levels. Western blot analysis was performed to measure G1 phase-related protein CDK4, CDK6, and cyclin D1. GAPDH was used as the internal control. Data are expressed as the mean ± standard error of the mean. CONT, control; ESCC, esophageal squamous cell carcinoma; PI, propidium iodide; n.s., no significant. *, compared to the control; #, compare with UA plus DIM combination treatment. * or #, p < 0.05; ** or ##, p < 0.01.
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