title: [Construction of bicistronic eukaryotic vector containing basic fibroblast growth factor and study of their functions in gene therapy for hearing impairment].
abstract: OBJECTIVE
  To construct basic fibroblast growth factor(bFGR) and enhance green fluorescence protein(EGFP) fusion gene eukaryotic expression vector internal ribosome entry site (pIRES)-bFGF-GFP and to evaluate the effect of transduction bFGF gene on noise induced hearing-loss in inner ear hair cells of guinea pigs.
  
  
  METHODS
  Human bFGF cDNA was inserted into mammalian expressed plasmid pIRES-EGFP. The recombinant expression plasmid pIRES-bFGF-EGFP was transfected into inner ear of guinea pigs, using lipofectin method. The transduced bFGF gene was mediated by SA lipidsome. IRES- bFGF-GFP was administered into the round window as rescue agent at the same time of noise exposure or as a protective agent 7 days before.
  
  
  RESULTS
  SA liposome-mediated bFGF expressed at a high level in the cochlea of guinea pigs, and in the rescue group, a significant lower hearing thresholds was displayed. pIRES- bFGF-EGFP could protect hair cells. It demonstrated that pIRES- bFGF-GFP could protect the inner ear both structurally and functionally. bFGF/EGFP gene could be transcripted and translated into inner ear hair cells of guinea pig. bFGF/EGFP gene could express a specific protein. The recombinant bFGF/EGFP had significant protective effect as well as manifestation of autonomous fluorescence.
  
  
  CONCLUSIONS
  bFGF/EGFP fusion protein not only expressed in hair cells of guinea pig but also showed significant bFGF activity and autonomous fluorescence. IRES induced exogenous gene could enter the hair cells.
authors.affiliation: Institute of Otorhinolaryngology, PLA General Hospital, Beijing 100853, China. shient@sina.com
authors.forename: Li
authors.initials: L
authors.lastname: Shi
authors.forename: Ming-min
authors.initials: MM
authors.lastname: Dong
authors.forename: Wen-jie
authors.initials: WJ
authors.lastname: Shi
authors.forename: Yin-yan
authors.initials: YY
authors.lastname: Hu
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.forename: Suo-qiang
authors.initials: SQ
authors.lastname: Zhai
authors.forename: Wei-yan
authors.initials: WY
authors.lastname: Yang
mesh_terms: D000818:Animals; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006168:Guinea Pigs; D006198:Hair Cells, Auditory; D006317:Hearing Loss, Noise-Induced / Q000628:therapy*; D006801:Humans; D008297:Male
chemical_list: D016222:Fibroblast Growth Factor 2