Fig 2: Tan IIA enhances the inhibitory effect of IM on tumor growth in TIB-152 ×enograft mice. Mice (n=5/group) were injected with TIB-152 cells. Subsequently, IM and/or Tan IIA were administered to the animals for 3 weeks. (A) Isolated tumors were obtained after drug treatment was completed. (B) Tumor growth curve was recorded over 21 days after treatment. (C) TUNEL images of tumor samples (scale bar, 50 µm) and (D) quantitative analysis of apoptosis rates. Immunohistochemistry images following staining with (E) Ki67 and (F) cleaved caspase-3 (scale bar, 50 µm), and (G) quantification of the results. **P<0.01 vs. TIB-152; ##P<0.01 vs. IM (5 µM); &&P<0.01 vs. Tan (20 µM). IM, imatinib; Tan IIA, tanshinone IIA; p-, phosphorylated. Tan IIA enhances the inhibitory effect of IM on tumor growth in TIB-152 ×enograft mice. Mice (n=5/group) were injected with TIB-152 cells. Subsequently, IM and/or Tan IIA were administered to the animals for 3 weeks. Immunohistochemistry images following staining with (H) VEGF and (I) MMP-9 (scale bar, 50 µm), and (J) quantification of the results. (K) Western blot images and (L) quantification of p-PI3K, PI3K, p-AKT, AKT, p-mTOR and mTOR protein levels. **P<0.01 vs. TIB-152; ##P<0.01 vs. IM (5 µM); &&P<0.01 vs. Tan (20 µM). IM, imatinib; Tan IIA, tanshinone IIA; p-, phosphorylated.

Fig 3: Tan IIA reverses the effect of the PI3K pathway activator IGF-1 on TIB-152 cells. Following pretreatment with IGF-1 for 24 h, the cells were treated with IM (5 µM), or IM (5 µM) plus Tan IIA (20 µM), for 48 h. (A) Western blot images and (B) quantification of p-PI3K, PI3K, p-AKT, AKT, p-mTOR, mTOR, Ki67, cleaved caspase-3, VEGF and MMP-9 protein levels. (C) Cell proliferation was detected by CCK-8 assay. (D) Comparison of apoptotic rates determined by flow cytometry. (E) Quantitative analysis of cell invasion based on Transwell assays. (F) Quantitative analysis of cell migration based on wound healing assays. **P<0.01 vs. TIB-152; ##P<0.01 vs. IGF-1 (10 µM); and $$P<0.01 vs. IM + IGF-1. IM, imatinib; Tan IIA, tanshinone IIA; IGF-1, insulin-like growth factor-1; p-, phosphorylated.

Fig 4: Paeonol inhibits the activation of the AKT signaling pathway. The SGC-7901 cell lines were treated with 0, 0.1, 0.2 and 0.4 mg/ml paeonol, and the expression levels of PI3K-Akt were detected by western blot analysis. *P<0.05, **P<0.01, ***P<0.001 vs. 0 mg/ml Pae. Pae, paeonol; PI3K, phosphoinositide 3-kinase; p-, phosphorylated.

Fig 5: Cryptotanshinone (CPT) inhibits the growth of Huh7 cell xenografts in nude mice. (A) Photograph of extracted tumors from each group of mice (n = 6). Statistical analysis of tumor volumes (C), body weight (D), and tumor weight (E) in each group of mice (n = 6). (B) TUNEL assays were conducted to measure apoptosis in tumor tissues using immunochemistry and immunofluorescence analyses. (F) The binding mode of CPT with phosphatidylinositol 3-kinase (PI3K) determined by molecular docking simulation. CPT formed stable hydrogen bonds with PI3K at SER-854 and VAL-851. (G) Electrostatic surfaces of CPT and PI3K obtained using Pymol. Lower electrostatic potential denotes better binding ability. (H) Tumor tissue lysates were extracted from the control and CPT-treated groups and the expression levels of PI3K, p-PI3K, AKT, p-AKT, mTOR, p-mTOR, PARP, cleaved PARP, caspase-3, cleaved caspase-3, Bcl-2, Bax, LC3-I, LC3-II, p62/SQSTM1, and Beclin1 proteins were assessed by western blotting.