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. 2021 Jan 15;17(2):589–602. doi: 10.7150/ijbs.49514

Figure 4.

Figure 4

Rhein synergistically enhances the therapeutic effects of oxaliplatin in PC cells via suppression of PI3K/AKT signaling. (A) Panc-1, MIAPaCa-2 and HPDE cells were treated with indicated concentration of oxaliplatin and Rhein for 24 or 48 h. CCK-8 assays were performed to determine the cell viability. (B) Combination Index (CI) values for oxaliplatin and Rhein in Panc-1 and MIAPaCa-2 cells were constructed by CalcuSyn software. CI < 1.0 indicates synergistic effect. (C) Panc-1 and MIAPaCa-2 cells were treated with oxaliplatin (25 μM) or Rhein (50 μM) alone or in combination for 24 h and allowed to form colonies for 14 days. Representative images are shown. Each bar represents means ± SD from three independent experiments. ***p < 0.001. (D) Western blot analysis of PARP, caspase-9, cleaved-caspase-3, Bcl-2 and IAP family proteins levels in Panc-1 and MIAPaCa-2 cells after treated with indicated concentration of oxaliplatin and Rhein. GAPDH was used as loading control. (E) Western blot analysis of PI3K/AKT pathway proteins treated with oxaliplatin (25 μM) or combination of oxaliplatin (25 μM) and Rhein 50 μM) for indicated times in Panc-1 or MIAPaCa-2 cells. GAPDH was used as loading control. (F) Panc-1 and MIAPaCa-2 cells were pretreated with 740Y-P or LY294002 for 1 h, followed by combination of oxaliplatin (25 μM) and Rhein (50 μM) treatment for 24 h. Cell viability was measured by CCK-8 assay. Each bar represents means ± SD from three independent experiments. *p < 0.05.