ECHS1Involved In Promoting The Proliferation And Anti-apoptosis In HepG2Cells

Objective:To observe the effect of the enoyl-CoA hydratase short chain1(ECHS1) onproliferation and apoptosis in HepG2cells, in order to provide a new theoretical basisof gene therapy and diagnosis to hepatocellular carcinoma.Methods:The constructed plasmid pGpU6/GFP/siRNA-ECHS1for ECHS1geneinterference was transfected into HepG2cells. The HepG2cells with stable ECHS1gene interference were screened by puromycin. The expression level of ECHS1inHepG2cells was detected by Real time PCR and Western blot. The proliferativeability of HepG2cells after transfection with pGpU6/GFP/siRNA-ECHS1wasdetected by CCK-8(Cell Counting kit-8) assay and BrdU (5-bromo-2’-deoxyuridine)assay. The apoptosis of HepG2cells transfected with pGpU6/GFP/siRNA-ECHS1was detected by flow cytometry and TUNEL assays. The expressions of ERK, P-ERK,Cyclin D1, Cyclin D3, Akt, P-Akt, GSK-3β, Bax and Bcl-xl were detected viaWestern blot.Results:The plasmid pGpU6/GFP/siRNA-ECHS1for ECHS1gene was successfullyconstructed. The expressions of ECHS1in HepG2cells after transfection withpGpU6/GFP/siRNA-ECHS1were significantly lower than those in the blank controlcells (HepG2cells without transfection) and the negative control cells (HepG2cellstransfected with pU6vector)(P<0.05). As compared with the negative controlcells, the proliferative ability was significantly inhibited detected by CCK-8andBrdU assays (P<0.05). As compared with the negative control cells, the expressionlevels of P-ERK, Cyclin D1and Cyclin D3proteins were lower in HepG2cells aftertransfection with pGpU6/GFP/siRNA-ECHS1, the expression level of Bax washigher than that in negative control cells (P<0.05). Apoptosis ratios of the HepG2 cells after transfection with pGpU6/GFP/siRNA-ECHS1were significant higher thanthose of negative control group by Flow cytometry and TUNEL assays (P<0.05). Ascompared with the negative control cells, the expression levels of P-Akt and Bcl-xlproteins were lower in HepG2cells after transfection with pGpU6/GFP/siRNA-ECHS1, the expression level of Bax was higher than that in negativecontrol cells (P<0.05).Conclusion:ECHS1promotes the proliferative ability of HepG2cells via regulatingERK-mediated cell signaling pathways and inhibits apoptosis of HepG2cells throughAkt and mitochondrial pathway.