Objective:To explore the molecular regulatory mechanism of down-regulation of E-cadherin inducing GSK-3β phosphorylation deactivation in hepatocellular carcinoma (HCC). Methods:The stable cell line without E-cadherin expression was constructed by RNA interference method. The expression levels of E-cadherin,Akt,p-Akt,GSK-3β and p-GSK-3β were examined by Western blot analysis;The expression levels of p-GSK-3β in HepG2-E-cadherin-shRNA stable cell line were examined by Western blot analysis after 10 μmol/L LY294002 (PI3K inhibitors) treatment;The mRNA level of Egr-1 and PTEN in HepG2-E-cadherin-shRNA cells were tested by RT-PCR;The expression of PTEN was also examined by Western blot analysis. Results:Compared with the blank,E-cadherin expression level down-regulated 82.54%,and the phosphorylation level of GSK-3β and Akt upregulated 298.93% and 218.98%,respectively. With the stimulation of PI3K inhibitor (LY294002),the phosphorylation level of GSK-3β down-regulated to 48.13% when compared with the blank. The mRNA level of PTEN dropped to 76.14%,and Egr-1 to 66.89% when compared with the blank. The expression of PTEN protein also down-regulated to 53.77% when compared with the blank. Conclusion:Down-regulation of E-cadherin deactivated GSK-3β via PI3K/Akt signaling in HCC cells.