Amarogentin Inhibits Liver Cancer Cell Angiogenesis after Insufficient Radiofrequency Ablation via Affecting Stemness and the p53-Dependent VEGFA/Dll4/Notch1 Pathway

Figure 4

Amarogentin inhibits iRFA-induced angiogenesis via the p53-dependent VEGFA/Dll4/Notch1 pathway in Huh7 cells. (a–c) The supernatant levels of VEGFA, the mRNA levels of CD133 and VEGFA, and the protein and phosphorylation levels of p53, Dll4, and Notch1 in iRFA cells treated with amarogentin, iRFA cells transfected with p53-shRNA, and iRFA cells transfected with p53-shRNA before treatment with amarogentin were detected by ELISA, RT-PCR, and WB assays, respectively. (d) The percentages of LCSCs in normal liver cancer cells, iRFA cells, iRFA cells treated with amarogentin, iRFA cells transfected with p53-shRNA, and iRFA cells transfected with p53-shRNA before treatment with amarogentin were indicated by CD133-PE staining and detected by flow cytometry. (e) HUVECs were cultured with supernatant from normal liver cancer cells, iRFA cells, iRFA cells treated with amarogentin, iRFA cells transfected with p53-shRNA, and iRFA cells transfected with p53-shRNA before treatment with amarogentin (400x). N = normal liver cancer; iRFA = iRFA cells; iRFA+A = iRFA cells with amarogentin treatment group; iRFA+shRNA = iRFA cells iRFA cells transfected with p53-shRNA group; iRFA+shRNA+A = iRFA cells transfected with p53-shRNA before treatment with amarogentin group ().

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