Effects Of Ezrin Enhancer Knockout On Human Esophageal Carcinoma Cells

Objective: This thesis aims to investigate the effects of targeting knockout of the ezrin gene enhancer of human esophageal cancer cell with the CRISPR/Cas9 gene editing technology on the gene expression,MAPK pathway-related protein expression,cell proliferation and migration of human esophageal carcinoma cells.Methods: The CRISPR/Cas9 recombinant plasmids were co-transfected into human esophageal carcinoma Eca-109 cells in the prophase,and then the cell line Eca-C2 whose ezrin enhancer was screened out with purinomycin.The expression levels of ezrin mRNA and protein were detected through real-time fluorescent quantitative PCR and Western blotting technique respectively.The expression levels of MAPK-pathway-related proteins were detected with protein array technology.The effects of ezrin enhancer knockout on the proliferation and migration of Eca-109 cells were analyzed through WST-1 method and cell wound healing assay,respectively.Results: Compared with the control cells,the expression levels of ezrin mRNA and protein in ezrin enhancer knockout cells were significantly reduced(all P<0.05),the MAPK-pathway-related proteins were down-regulated in 17 tested proteins,and the cell proliferation and migration ability were significantly inhibited(all P<0.05).Conclusion: Caused by ezrin enhancer knockout,the down-regulation of ezrin gene expression can significantly inhibit the cell proliferation and migration of human esophageal carcinoma Eca-109 cells.