Ezrin enhancer knockout inhibits the proliferation and migration of human esophageal carcinoma Eca-109 cells
10.3872/j.issn.1007-385x.2019.01.006
- VernacularTitle:Ezrin增强子敲除可抑制人食管癌Eca-109细胞的增殖和迁移
- Author:
LEI Yue
1
;
YE Qingsong
1
;
WEI Jinqi
2
;
LI Wenna
1
;
MO Zhentao
2
;
ZHANG Qingfeng
1
;
GAO Shuying
3
Author Information
1. Department of Biochemistry and Molecular Biology
2. Zhuhai Premier-Discipline Enhancement Scheme of Pharmacology, Zhuhai Campus of Zunyi Medical University
3. Department of Gastroenterology, the Fifth Affiliated Hospital of Sun Yat-sen Universit
- Publication Type:Journal Article
- Keywords:
ezrin enhancer;
gene knockout;
esophageal carcinoma;
Eca-109 cell;
proliferation;
migration
- From:
Chinese Journal of Cancer Biotherapy
2019;26(1):29-35
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effects of ezrin enhancer knockout on ezrin gene expression, cell proliferation and migration of human esophageal carcinoma Eca-109 cells.
Methods: The CRISPR/Cas9 recombinant plasmids targeting upstream/downstream of human ezrin enhancer were co-transfected into human esophageal carcinoma Eca-109 cells, and the cell line Eca-C2 with ezrin enhancer knockout was screened by purinomycin. Then the expression levels of ezrin mRNAand protein in Eca-C2 cells were detected by Real-time quantitative PCR (qPCR) and Western blotting, respectively; The expression levels of MAPK-pathway-related proteins were detected by protein array technology; and the effects of ezrin enhancer knockout on the proliferation and migration of Eca-C2 cells were analyzed by WST-1 method and wound-healing assay, respectively.
Results:The human esophageal carcinoma cell line Eca-C2 with stable ezrin enhancer knockout was established successfully. Compared with control cells, the mRNA and protein expressions of ezrin in Eca-C2 cells were significantly reduced (all P<0.05).Among the 17 detected MAPK pathway related proteins in Eca-C2 cells, 9 proteins (AKT, CREB, GSK3b, MKK6, mTOR, P38, P53, P70S6K and RSK1) were down-regulated, and the cell proliferation and migration were significantly inhibited (all P<0.05).
Conclusion: ezrin enhancer knockout can significantly inhibit the cell proliferation and migration of human esophageal carcinoma Eca-109 cells.
- Full text:20190106.pdf
