In vitro study of joint intervention of E-cad and Bmi-Ⅰ mediated by transcription activator-like effector nuclease in nasopharyngeal carcinoma
10.11817/j.issn.1672-7347.2018.03.001
- VernacularTitle:类转录激活因子效应物核酸酶介导的E-cad和Bmi-Ⅰ基因联合干预鼻咽癌的体外研究
- Author:
Tingting LUO
1
;
Aifen YAN
;
Lian LIU
;
Hong JIANG
;
Cuilan FENG
;
Guannan LIU
;
Fang LIU
;
Dongsheng TANG
;
Tianhong ZHOU
Author Information
1. 暨南大学生命科学技术学院分子生物学教研室
- Keywords:
transcription activator-like effector nuclease;
E-cadherin;
B-lymphoma Moloney murine leukemia virus insertion region-1;
gene targeting;
nasopharyngeal carcinoma
- From:
Journal of Central South University(Medical Sciences)
2018;43(3):229-239
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of intervention of E-cadherin (E-cad) and B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1) mediated by transcription activator-like effector nuclease (TALEN) on the biological behaviors ofnasopharyngeal carcinoma cells.Methods:Multi-locus gene targeting vectors pUC-DS1-CMV-E-cad-2A-Neo-DS2 and pUC-DS1-Bmi-1 shRNA-Zeo-DS2 were constructed,and the E-cad and Bmi-1 targeting vectors were transferred with TALEN plasmids to CNE-2 cells individually or simultaneously.The integration of target genes were detected by PCR,the expressions of E-cad and Bmi-1 were detected by Western blot.The changes of cell proliferation were detected by cell counting kit-8 (CCK-8) assay.T-he cell cycle and apoptosis were detected by flow cytometry.The cell migration and invasion were detected by Transwell assay.Results:The E-cad and Bmi-1 shRNA expression elements were successfully integrated into the genome of CNE-2 cells,the protein expression level of E-cad was up-regulated,and the protein expression level of Bmi-1 was down-regulated.The intervention of E-cad and Bmi-1 didn't affect the proliferation,cell cycle and apoptosis of CNE-2 cells,but it significantly inhibited the migration and invasion ability of CNE-2 cells.Furthermore,the intervention of E-cad and Bmi-1 together significantly inhibited the migration ability of nasopharyngeal carcinoma cells compared with the intervention of E-cad or Bmi-1 alone (all P＜0.01).Conclusion:The joint intervention of E-cad and Bmi-1 mediated by TALEN can effectively inhibit the migration and invasion of nasopharyngeal carcinoma cells in vitro,which may lay the preliminary experimental basis for gene therapy of human cancer.
