Association Between Epithelial-Mesenchymal Transition and the VEGFa/VEGFR2 Autocrine Pathway in Human Hepatocellular Carcinoma Huh7 Cells and the Regulatory Effect of Cinobufagin
Objective To investigate the effect of Cinobufagin (CBG)Injection on the VEGFa/VEGFR2 autocrine signaling pathway and epithelial-mesenchymal transition in human hepatocellular carcinoma Huh7 cells.Methods CCK-8 assay,scratch assay,Transwell invasion assay,and sphere-forming assay were used to observe the proliferation,migration,invasion,and sphere formation abilities of Huh7 cells;double-label immunofluorescence was used for the colocalization of VEGFa and VEGFR2 in human hepatocellular carcinoma Huh7 cells;ELISA was used to measure the level of VEGFa in the supernatant of Huh7 cells,and Western blot was used to measure the expression levels of the proteins associated with the VEGFa/VEGFR2 pathway and epithelial-mesenchymal transition in Huh7 cells.Results CBG Injection significantly inhibited the proliferation,migration,invasion,and sphere formation abilities of human hepatocellular carcinoma Huh7 cells.The VEGFa/VEGFR2 auto-crine pathway might exist in Huh7 cells,and CBG Injection inhibited the secretion of VEGFa,downregulated the expression levels of downstream p-VEGFR2,p-PI3K,and p-AKT and the mesenchymal markers N-Cadherin,Vimentin,and Snail,and upregulated the protein expression levels of the epithelial marker E-Cadherin.Conclusion CBG Injection may regulate the epi-thelial-mesenchymal transition of Huh7 cells through the VEGFa/VEGFR2 autocrine pathway.
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