Impact Of ECEL1 Gene Knockdown On The Expression Of CDH1,EGR1,FOSL1 And MAP3K1 In Human Hepatocellular Carcinoma Cell

Objective:To investigate the effects of ECEL1 knock-down on the expression of CDH1,EGR1,FOSL1 and MAP3K1 in HHCC BEL-7404.To explore the mechanism of inhibiting proliferation and promoting apoptosis of HHCC by knock-down ECEL1 gene,and to provide a new target for the prevention and treatment of HHCC.Methods:In this experiment,experimental group(sh ECEL1 group)and control group(sh Ctrl group)were set up.The HHCC BEL-7404 was infected in vitro by the lentiviral vector of ECEL1;The efficiency of infection was measured by the fluoresce nce microscope.Total RNA was extracted from the two groups of cells;To detecting ECEL1 m RNA expression of BEL-7404 cells after Ecel1 knock-down by q PCR.The expression of CDH1,EGR1,FOSL1 and MAP3K1 in the HHCC BEL-7404 after ECEL1 knockdown was detected by q PCR.Results:(1)Infection of BEL-7404 lentivirus in two groups of HHCC: green fluorescence can be observed in sh ECEL1 and sh Ctrl,the calculated efficiency of infection for each group was above 80%,indicating successful infection of lentiviral vector in both groups.(2)Expression of ECEL1 m RNA in HHCC BEL-7404 in two groups after infection: the expression of ECEL1 m RNA in sh ECEL1 was inhibited in that its expression level significantly reduced(<0.01),and the efficiency of knock-down was 70.5 %,as compared to sh Ctrl.This indicates that the stable HHCC with ECEL1 knock-down was successfully constructed.(3)Expression of CDH1,EGR1,FOSL1 and MAP3K1 after ECEL1 gene knockdown in BEL-7404 cells: as compared to sh Ctrl,the expression level of CDH1 and MAP3K1 m RNA in the HHCC BEL-7404 in sh ECEL1 rose,while that of EGR1 m RNA reduced,the statistical processing was significant(<0.01).As compared to sh Ctrl,the expression level of FOSL1 m RNA in sh ECEL1 reduced,but no statistical difference(p>05.0).Conclusions:The expression of CDH1 and MAP3K1 genes in BEL-7404 cells are enhanced after ECEL1 knock-down,and the expression of EGR1 and Fosl1 genes are down-regulated,ECEL1 may regulate the proliferation and apoptosis of HHCC through the abovementioned pathway.