Effect of Beclin 1 RNA interference on Vit K3 injured SMMC-7721 cells

AIM: To observe the effect of Beclin 1 silencing by RNA interference (RNAi) technique to the injury of SMMC-7721 hepatoma cells by vitamin K3 (Vit K3).METHODS: The recombinant plasmid Psilencer 3.1-siRNA-Beclin 1 was transfected into SMMC-7721 hepatoma cells by eukaryotic cell transfection technique. Plasmid vector and cell culture medium were used as negative and control, respectively. The cells were collected 48 h later to extract cell RNA and total protein and to detect Beclin 1 gene expression by RT-PCR and Western blotting. 40 μmol/L Vit K3 was used to treate the Beclin 1-siRNA cells, Hoechst33342 staining was used for the determination of the percentage of cell apoptosis.RESULTS: Compared with the control group, the synthetic siRNA of Beclin 1 significantly decreased the levels of Beclin 1 mRNA and protein expressions. Beclin 1 mRNA was up-regulated in 40 μmol/L Vit K3 treated SMMC-7721 hepatoma cells, the percentage of apoptosis cells increased (P﹤0.01). In beclin 1-siRNA cells, Beclin 1 mRNA was down-regulated obviously, the percentage of apoptosis cells increased significantly compared with the 40 μmol/L Vit K3 group (P﹤0.01).CONCLUSION: The transfection of SMMC-7721 hepatoma cells by Psilencer3.1-siRNA-Beclin 1 effectively inhibits the expressions of Beclin 1 mRNA and protein, inhibits the activation of Beclin 1 dependent autophagic signaling pathway, and aggravates the apoptosis induced by Vit K3.