Effect Of ECEL1 Silencing On Expression Of DUSP1,THBS1,PTEN And EGR1 In Hepatocellular Carcinoma Cells

Objective: To study the effect of silencing ECEL1 on the expression of DUSP1,THBS1,PTEN and EGR1 in human liver cancer cells(BEL-7404).Methods:(1)Human liver cancer cells(bel-7404)were infected with lentivirus by ECEL1 gene RNA,and the control group was set up at the same time to observe the lentivirus infection efficiency of liver cancer cells under a fluorescence microscope.(2)qPCR was used to detect the expression levels of ECEL1,DUSP1,THBS1,PTEN and EGR1 mRNA in bel-7404 human hepatocellular carcinoma cells silenced by ECEL1.(3)WB was used to detect the expression levels of DUSP1,THBS1,PTEN and EGR1 proteins in bel-7404 human liver cancer cells silenced by ECEL1.Results:(1)After the lentivirus infected human liver cancer cells(bel-7404)for 72 hours,the fluorescence microscope showed that the liver cancer cells were in good condition and the infection efficiency reached more than 80%.The qPCR detection results showed that the silence rate of ECEL1 gene was up to 84.1%,and the mRNA expression of ECEL1 gene in shECEL1 group was significantly inhibited(p<0.05).(2)qPCR was used to detect the mRNA expression of relevant genes in liver cancer cells.The results showed that after ECEL1 was silenced,the expression of DUSP1,THBS1 and PTEN mRNA was up-regulated,and the expression of EGR1 mRNA was down-regulated.(3)WB technology was used to detect the expression of related gene proteins in liver cancer cells.The results showed that after ECEL1 was silenced,the expression of DUSP1,THBS1 and PTEN proteins was up-regulated,and the expression of EGR1 protein was down-regulated.Conclusion:Silence of ECEL1 gene can lead to up-regulation of DUSP1,THBS1 and PTEN gene expression and down-regulation of EGR1 gene expression in human liver cancer cells(BEL-7404).