title: [Specific inhibition of Raf-1 gene expression in HNE1 by RNA interference].
abstract: OBJECTIVE
  To silence the expression of Raf-1 gene in HNE1 cells using vector-based RNA interference (RNAi) technique.
  
  
  METHODS
  The vector containing the human U6 promoter was used for targeted gene silencing when a dsDNA oligonucleotide encoding an appropriate shRNA was ligated into the vector, and 67nt oligonucleotide fragment was inserted into the downstream of the U6 promoter. Plasmids containing different Raf-1 target sequences [ (1) pshuttle-Raf-1-a( 225), (2) pshattle-Raf-1-b ( 358) and (3) pshuttle-Raf-1-c(474)], were transfected into HNE1 cells. Expression of Raf-1 mRNA was assayed by RT-PCR. Apoptosis were determined by cytometry.
  
  
  RESULTS
  Vector-based RNAi had advantages over antisense RNA because it could be delivered to the target cell more efficiently, and effect could last longer. Raf-1 expression could be inhibited by plasmid-expressed shRNA. Three different targeting sequences were selected from Raf-1 gene, and the inhibitory effect of pSIREN shuttle-Raf-1-b (358) was biggest.
  
  
  CONCLUSIONS
  Raf-1 expression in HNE1 cells can be inhibited significantly using plasmid-based RNAi.
authors.affiliation: Department of Otorhinolaryngology, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Li-Zhong
authors.initials: LZ
authors.lastname: Liu
authors.forename: Chuan-Yu
authors.initials: CY
authors.lastname: Liang
authors.forename: Yan-Ling
authors.initials: YL
authors.lastname: Dou
authors.forename: Ling
authors.initials: L
authors.lastname: Tian
authors.forename: Yu-Quang
authors.initials: YQ
authors.lastname: Wei
authors.forename: Yan-Jun
authors.initials: YJ
authors.lastname: Wen
authors.forename: Jiong
authors.initials: J
authors.lastname: Li
authors.forename: Hong-Xing
authors.initials: HX
authors.lastname: Deng
authors.forename: Bing
authors.initials: B
authors.lastname: Kan
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D045744:Cell Line, Tumor; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D011401:Promoter Regions, Genetic; D019908:Proto-Oncogene Proteins c-raf / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D019908:Proto-Oncogene Proteins c-raf