Study On The Effect And Mechanism Of LKB1 With The Relationship Of EMT In Hepatocellular Carcinoma Cells

ObjectiveTo Study on the effect and mechanism of LKB1 on hepatocellular carcinoma cells(HCC) and it’s function of the epithelial-mesenchymal transition(EMT) in HCC. MethodsProtein expressions of LKB1 in HCC cell line SNU449 and liver cell line HL7702 were detected by western blotting.The experiment was divided into three groups:Control group and NC-shRNA,SNU449 cells transfected with LKB1-shRNA.To detect the cell activity by MTT.Real-time quantitative RT-PCR was used to investigate the mRNA expression of LKB1,E-cadherin and N-cadherin after transfection in compared with the control group and NC-shRNA group.Western blot was used to detect the expression LKB1、E-cadherin and N-cadherin after transfection in compared with the control group and NC-shRNA group. Results1.The expressions of LKB1 by western blot in HCC cell line SNU449 and liver cell line HL7702 was 0.58±0.13 and 0.86±0.11,which in cell line SNU449 was significantly lower than in HL7702(P<0.05).2.The level of expressions of LKB1 mRNA in the control group and NC-shRNA group,there is no obvious change.But the level of expressions of LKB1 mRNA with LKB1-shRNA transfected with was elevated in compare with the NC-shRNA group(P<0.05). And by western blot,the expression of LKB1 protein in control group,NC-shRNA group and LKB1-shRNA group was:0.55±0.11,0.51±0.07,0.28±0.13.There is no obvious change in control group and NC-shRNA group,but LKB1-shRNA group is lower than NC-shRNA group(P<0.05).3.The cell activity detect by MTT after 1d,2d,3d,4d,5d,the Control group and the NC-shRNA group,there is no clear change,but the LKB1-shRNA group was significantly higher than the NC-shRNA(P<0.05).4.The mRNA expression of E-cadherin and N-cadherin by RT-PCR,which Control and NC-shRNA group,there is no clear change.But the E-cadherin in LKB1-shRNA group was lower than NC-shRNA group,the N-cadherin level was higher than NC-shRNA group(P<0.05). By western blot,the expression E-cadherin protein in Control and NC-shRNA group,there is no obvious difference,but the level of E-cadherin was declined than NC-shRNA group,the N-cadherin level was elevated han NC-shRNA group(P<0.05). Conclusions1.The expression level of LKB1 elevated significantly in liver cell with HCC.2.Silencing LKB1 promotes proliferation of HCC.3.LKB1 is involved in EMT regulation in HCC.