title: [Establishment of a stable nasopharyngeal carcinoma cell line with lentivirus-mediated RNA interference for EIF4G1 gene silencing].
abstract: OBJECTIVE
  To establish a nasopharyngeal carcinoma (NPC) cell line with stable EIF4G1 gene silencing induced by small interfering RNA (siRNA).
  
  
  METHODS
  The EIF4G1 mRNA levels in 8 NPC cell lines including 5-8F, 6-10B, C666-1, CNE1, CNE2, HNE1, HONE1, and SUNE1 were detected by fluorescence quantitative RT-PCR (QRT-PCR). The recombinant lentivirus shRNA expression plasmid targeting EIF4G1 gene was packaged into mature lentivirus by 293FT cells and used to infect 5-8F cells. After blasticidin selection of NPC cells with constant expression of the EIF4G1-siRNA, the efficiency of EIF4G1 mRNA expression interference was determined using QRT-PCR.
  
  
  RESULTS
  The 8 NPC cell lines showed differential expression of EIF4G1 mRNA, among which 5-8F cells had the highest EIF4G1 expression. The recombinant lentivirus plasmid pLenti6/BLOCK-iT-DEST/EIF4G1-shRNA was successfully constructed and verified by PCR and sequencing. The EIF4G1 mRNA level of 5-8F cells infected with shRNA-EIF4G1 lentivirus was significantly reduced as compared with the negative control and the blank control cells.
  
  
  CONCLUSIONS
  The recombinant lentivirus vector pLenti6/BLOCK- iT-DEST/EIF4G1-shRNA we constructed results in marked downregulation of EIF4G1 mRNA expression and constant expression of EIF4G1-siRNA after infection of 5-8F cells.
authors.affiliation: Cancer Research Institute, Key Lab for Transcriptomics and Proteomics of Human Major Diseases, Southern Medical University, Guangzhou 510515, China.
authors.forename: Lu-xia
authors.initials: LX
authors.lastname: Tu
authors.forename: Wei-yi
authors.initials: WY
authors.lastname: Fang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Liu
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Ying
authors.initials: Y
authors.lastname: He
authors.forename: Si-ming
authors.initials: SM
authors.lastname: Xie
authors.forename: Kai-tai
authors.initials: KT
authors.lastname: Yao
mesh_terms: D045744:Cell Line, Tumor; D039603:Eukaryotic Initiation Factor-4G / Q000096:biosynthesis / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: C113920:EIF4G1 protein, human; D039603:Eukaryotic Initiation Factor-4G; D012333:RNA, Messenger; D034741:RNA, Small Interfering