AEBP2 Promotes Proliferation,Migration And Invasion Of Hepatocellular Carcinoma Cells Through PI3K/Akt Signaling Pathway

Objective:To investigate the effects of adipocyte enhancer binding protein 2(AEBP2)on the proliferation,apoptosis,migration and invasion of hepatocellular carcinoma cells(HCC)and the effect of PI3K/Akt signaling pathway.Methods:The UALCAN database was used to analyze the expression of AEBP2 in HCC samples,and kaplan-Meier Plotter database was used to analyze the prognostic correlation between AEBP2 and HCC patients.Real-time PCR and Western blotting were used to detect the expression levels of AEBP2 mRNA and protein in normal liver cells WRL68 and HCC HepG2 and Huh-7 cells.Silencing the expression of AEBP2 in HepG2 and Huh-7 cells,the effect of cell proliferation,apoptosis,migration and invasion were detected by plate cloning,cell proliferation,flow cytometry,wound-healing assay and cell invasion assay.Immunofluorescence and Western blotting were used to detect the expression of epithelial-mesenchymal transition(EMT)-related proteins after AEBP2 silencing.The expression data of patients with HCC was downloaded from TCGA database.The data were divided into two groups according to the expression of AEBP2 and used to identify differently expression genes,and the pathway enrichment of the differential genes was performed.After the PI3K pathway was activated by IGF-1,the abilities of migration and invasion in HCC HepG2 and Huh-7 cells were detected by Would-healing assay and cell invasion assay,and the expression of PI3K/Akt pathway-related proteins were detected by Western blotting.Results:The expression of AEBP2 was significantly up-regulated in HCC,and HCC patients with AEBP2 high expression had lower overall survival rate(P<0.05).The mRNA and protein levels of AEBP2 in HepG2 and Huh-7 cells were higher than WRL68 cells(P < 0.05).AEBP2 silenced HepG2 and Huh-7 cell lines were successfully constructed.Compared with the control group,down-regulation of AEBP2 expression inhibited the proliferation,migration and invasion of HCC HepG2 and Huh-7 cells,as well as promoted apoptosis(all P<0.05).Silencing AEBP2 inhibited EMT in HCC HepG2 and Huh-7 cells,up-regulated the expression of E-cadherin protein,down-regulated the expression of N-cadherin and Snail2 proteins(all P<0.05).Differently expression genes between high and low AEBP2 group were enriched in the PI3K/Akt pathway.After silencing AEBP2,the protein expression levels of downstream PI3K signaling pathway members were significantly down-regulated,such as p-Akt(S473),mTOR,MMP-2,MMP-9(all P<0.05).IGF-1could partially reversed the inhibitory effect of down-regulated AEBP2 on migration and invasion of HepG2 and Huh-7 cells.Conclusions:AEBP2 is highly expressed in HCC HepG2 and Huh-7 cells;AEBP2 may promote the proliferation,migration and invasion of HCC HepG2 and Huh-7 cells,and inhibits cell apoptosis through the PI3K/Akt signaling pathway.