检验医学与临床Issue(7)：888-890,894,4.DOI:10.3969/j.issn.1672-9455.2015.07.005
慢病毒感染稳定敲减eIF4E基因的鼻咽癌CNE2细胞系的建立
Construction of stable eIF4E knockdown nasopharyngeal carcinoma CNE2 cell line by lentivirus infection
摘要
Abstract
Objective To establish a nasopharyngeal carcinoma CNE2 cell line with stable interference of eu‐karyotic initiation factor 4E (eIF4E) gene .Methods The interference sequences ,targeting eIF4E gene sequence ,were designed ,coated and transfected into CNE2 cells ,by using lentiviral vector infection system based on pGC‐LV .The cells containing stable transformants were screened by the ability of resistance to puromycin .The expression of eIF4E was assayed by real‐time polymerase chain reaction and Western Blot .The effects of eIF4E knockdown on prolifera‐tion and migration of CNE2 cells were tested .Results The lentivirus particles with short hairpin shRNA targeting eIF4E gene were constructed and screened successfully .CNE2 cells were transfected and anti‐pure positive cell ,which was named CNE2siEp cell ,were obtained by antibiotic screening .Compared with negative control cells (CNE2siNC) , the expression of eIF4E mRNA and protein in CNE2siEp cells were significantly decreased by 76 .0% and 50 .0% re‐spectively .The proliferation and migration of CNE2siEp cells were also decreased by 85 .0% and 59 .0% respective‐ly .Conclusion The stable eIF4E knockdown cell line (CNE2siEp) was constructed successfully ,in which the expres‐sion and functions of eIF4E gene were inhibited significantly .关键词
真核起始因子4E/敲减/慢病毒感染/RNA干扰Key words
eukaryotic initiation factor 4E/knockdown/lentivirus infection/RNA interference引用本文复制引用
赵毅,王燕,胡新荣..慢病毒感染稳定敲减eIF4E基因的鼻咽癌CNE2细胞系的建立[J].检验医学与临床,2015,(7):888-890,894,4.基金项目
广东省卫生厅医学科研基金项目（B2013291）。 （）
