Emodin attenuates high lipid-induced liver metastasis through the AKT and ERK pathways in vitro in breast cancer cells and in a mouse xenograft model

Figures

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  Fig. 1. Results of Reactome enrichment. (A) Overview diagram. The font size and color are consistent with figure C. (B) Sankey's diagram of the enrichment pathways and their categories. (C) Pie chart of the enrichment analysis results. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

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  Fig. 2. Results of GO, KEGG and Reactome enrichment analyses. (A) The overview results of enrichment were analyzed by g:Profiler. (B) Top 10 terms in GO enrichment pathways by WebGestalt. (C) Top 10 terms in KEGG enrichment pathways by WebGestalt. (D) Top 10 terms in REAC enrichment pathways by WebGestalt. Pathways of lipid metabolism are highlighted in red. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

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  Fig. 3. PPIs and interactions of TF-miRNAs by NetworkAnalyst analysis. (A) PPIs. (B) TF-miRNA interactions. (C) Signaling pathway interactions.

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  Fig. 4. Emodin attenuated the high potential malignancy of cancer cells coincubated with HFD liver homogenate extract. After the animals were sacrificed by anesthesia and bled, the mouse livers were harvested, and liver homogenates were prepared. Homogenates from the same treatment groups were mixed for subsequent cell experiments. For details of the determination, please refer to the Methods section. Data are shown as means ± SDs (n = 3). *P < 0.05 and **P < 0.01 compared to the corresponding vehicle group. #P < 0.05 and ##P < 0.01 compared to the LFD Veh group. Three biological replicates were used for each treatment group. Emodin inhibited cancer cell migration based on wound healing assays in MDA-MB-231 cells (A/E) and SKBR3 cells (C/E) and invasion as determined by transwell invasion assays in MDA-MB-231 cells (B/F) and SKBR3 cells (D/F). Emodin reduced tumor cell activity, as shown by CCK-8 analysis (G), and proliferation, as shown by a colony formation assay (H).

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  Fig. 5. Emodin inhibited breast cancer liver metastasis by regulating lipid synthesis in mice. Mice were fed a HFD or LFD for 9 weeks and were given an intravenous injection of luciferase-labeled MDA-MB-231 cells (4.0 × 106 cells/mouse) after anesthesia with pentobarbital sodium (50 mg/kg) (n = 6/group). The mice received emodin (400 mg/kg, p. o., q. d.) or vehicle for 4 weeks. For in vivo tumor proliferation, blood glucose and blood lipid detection in tumor-bearing mice, please refer to the Methods section. The means ± SDs with an asterisk (*) are significantly (P < 0.05) different from the vehicle. (A) Emodin attenuated breast cancer liver metastasis in mice. (B) Tumor proliferation in the liver of tumor-bearing mice. Effects of emodin on liver weight (C), liver TG (D), and liver FFA levels (E) in mice. (F) Emodin inhibited fatty acid synthesis and cholesterol synthesis in HepG2 cells. Three biological replicates were used for each treatment group. Metformin (Met) at a concentration of 1 mmol/L was used as a positive control. (G) Emodin reduced the lipogenic regulator Serbp1c and TG synthesis in the liver.

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  Fig. 6. Emodin regulated the ERK/AKT signaling pathway and interacted with CSNK2A1, ESR, PIM1 and PTP4A3. (A) The molecular structure of emodin. The binding action and binding site of emodin to CSNK2A1 (B), ESR1 (C), ESR2 (D), PIM1 (E) and PTP4A3 (F). (G) The results of the binding affinity between emodin and its targets. (H) Emodin decreased the phosphorylation levels of ERK1/2 and AKT, as shown by western blotting analysis. Three biological replicate was used for each treatment group. *p < 0.05 compared with the Veh group. (I) Schematic diagram of emodin inhibiting breast cancer liver metastasis.

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