Study On Cell Cycle And Proliferation Of Hepatocellular Carcinoma Cells Be Influenced By EEF1A1

Objective: To investigate the effects of RNA interference on eukaryotic translation elongation factor 1A1(eEF1A1)on the proliferation of hepatocellular carcinoma Hep3 B cells and to explore its possible mechanism.Methods: A recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA in which the short hairpin RNA(shRNA)targeting eEF1A1 was established,and then it was transfected into the Hep3 B cells.The efficacy of gene silencing was confirmed by Real-Time PCR and Westernblotting.The cell growth ability,colony formation ability,and cell cycle distribution of Hep3 B cells infected with pGPU6/GFP/Neo-eEF1A1-shRNA were detected by cell counting kit-8(CCK-8)assay,colony formation assay,and flow cytometry,respectively.The cyclin D1 and cyclin-dependent kinase 4(CDK4)protein expressions were examined by Westernblotting.Results: The recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA was successfully constructed,and the stably transfected cells were established.As compared with negative control(NC)cells which were tranfected with pGPU6/GFP/Neo-NC vector,the abilities of cell growth ability and colony formation ability of the cells which were tranfected with pGPU6/GFP/Neo-eEF1A1-shRNA were obviously decreased(P<0.05),the percentage of the cells in G1-phase was increased and which in S1-phase was decreased(P<0.05),as well as the expression levels of Cyclin D1,CDK4 proteins were down-regulated(P<0.05).Conclusion: Down-regulation of eEF1A1 expression can inhibit the proliferation of Hep3 B cells,this results suggests that over-express eEF1A1 may be related to the proliferation of hepatocellular carcinoma.