Effect Of ECEL1 Knockdown On Apoptosis Of Hepatocellular Carcinoma Cells

Objective: To study the effect of ECEL1 gene on the apoptosis of hepatocellular carcinoma cells,and provide a new target for the prevention and treatment of hepatocellular carcinoma.Methods:The lentivirus vector of ECEL1 gene was used to infect human hepatocellular carcinoma cells(BEL-7404,Huh-7)in vitro and the infection efficiency was detected by fluorescence microscopy.Total RNA and protein were extracted,and the expression of ECEL1 and protein was detected by q-PCR and Western blot.The apoptotic rate and cell cycle of BEL-7404 and Huh-7 cells were detected by flow cytometry.Finally,Caspase-Glo 3/7 Assay kit was used to detect the activity of Caspase 3/7in BEL-7404 and Huh-7 cells.Results:1.After the lentivirus vector of ECEL1 gene was infected with human hepatocellular carcinoma cells in vitro,the detection efficiency was more than 80%.2.The expression of ECEL1 in BEL-7404 and Huh-7 cells in shECEL1 group was significantly lower than that in shCtrl group(P < 0.01).3.Flow cytometry showed that the apoptotic rate ofBEL-7404 and Huh-7 cells increased significantly after ECEL1 gene knockdown,which was significantly higher than that of shCtrl control group(P < 0.01).4.After shRNA inhibited the expression of ECEL1 gene in BEL-7404 and Huh-7 cells,the S-phase of BEL-7404 cells decreased significantly,while the number of G2/M cells increased significantly(P <0.01),while the S-phase,G1-phase and G2/M-phase cells of Huh-7 cells did not change significantly.5.Caspase-Glo < 3/7 Assay kit test showed that the activity of Caspase-3/7 of BEL-7404 and Huh-7 cells in shECEL1 group was significantly higher than that in shCtrl control group(P < 0.01).Conclusions:1.Knockdown of ECEL1 gene can promote the apoptosis of hepatocellular carcinoma cells(BEL-7404,Huh-7).