(A) Proteins interacted with AKT3 and YWHAZ was predicted by String database (http://string-db.org). (B) FOXO3a protein expression in HCC patients according to Edmondson-Steiner grade and vascular invasion. (C) FOXO3a protein expression showed a significant positive correlation with miR-22 and a significant inverse correlation with YWHAZ protein expression in HCC tissues. (D) HCCLM9 cells were transfected with miR-22 mimic or pcDNA3.1-YWHAZ recombinant plasmid. 48 h later, protein expression of AKT, pAKT, YWHAZ, pFoxO3a, FOXO3a in different cellular components were detected by western blotting. (E) FOXO3a mRNA expression in response to miR-22 mimic and YWHAZ overexpression was determined by qRT-PCR. (F) and (G) In some cases, HCCLM9 cells were pretreated with LY294002 (PI3K inhibitor, 20 μM) for 1 h, and then transfected with miR-22 mimic or YWHAZ-expressing plasmid for 48 h. mRNA and protein expressions of E-caherin, N-cadherin, MMP2 and MMP9 were detected by qRT-PCR (F) and western blotting (G). Data are displayed as the Mean ± SD of three independent experiments.