ABSTRACT
Objective: To investigate the effect of ECHS1 (enoyl-coenzyme A hydratase, short chain 1) on the proliferation of hepatocellular carcinoma HepG2 cells. Methods: The recombinant plasmid pGPU6/GFP/siRNA-ECHS1 for ECHS1 gene interference was transfected into HepG2 cells. The HepG2 cells with stable ECHS1 gene interference were screened by puromycin. The expression level of ECHS1 protein in HepG2 cells was detected by Western blotting. The proliferative ability of HepG2 cells after transfection with pGPU6/GFP/siRNA-ECHS1 was detected by CCK-8 (cell counting kit-8) assay and BrdU (5-bromo-2'-deoxyuridine) assay. The expression levels of ERK (extracellular signal regulated kinase), p-ERK (phosphorylated-ERK), cyclin D3 and cyclin D1 were detected by Western blotting. Results: The HepG2 cells with stable ECHS1 gene interference were successfully established. The expression level of ECHS1 protein in HepG2 cells after transfection with pGPU6/GFP/siRNA-ECHS1 was significantly lower than that in the blank control cells (HepG2 cells without transfection) and the negative control cells (HepG2 cells transfected with pGPU6 vector) (P < 0.05). As compared with the negative control cells, the proliferative ability was significantly inhibited (P < 0.05) and the expression levels of p-ERK, cyclin D3 and cyclin D1 proteins were lower in HepG2 cells after transfection with pGPU6/GFP/siRNA-ECHS1 (P < 0.05). Conclusion: ECHS1 may promote the proliferative ability of HepG2 cells via enhancing the expression levels of p-ERK, cyclin D3 and cyclin D1. Copyright © 2013 by TUMOR.