title: [In vitro experimental study of killing Hep-2 cells of laryngeal cancer with suicide fusion gene CDglyTK].
abstract: OBJECTIVE
  To study the killing effect of suicide gene CDglyTK combined with GCV or 5-FC on the human laryngeal carcinoma Hep-2 cell line in vitro.
  
  
  METHODS
  Constructed plasmid pcDNA3.1 (-) CMV. CDglyTK was verified by enzyme digestion of Xho I /Hind III and automatic sequence analysis, then it was introduced into Hep-2 cells by electroporation to yield cells expressing CDglyTK stably after selecting with G418(400 ng/L) for 14 da. The expression of CDglyTK mRNA in transfected Hep-2 cells was tested by RT-PCR. Compared with Hep-2 cells transferred with pcDNA3.1(-), in vitro chemosensitivity of CDglyTK-expressing Hep-2 cells to 5-FC, GCV or 5-FC + GCV was detected by MTT assay.
  
  
  RESULTS
  The recombinant plasmid contained full-length coding region sequence of CD and TK gene. A anticipated 707 bp fragment was amplified from total RNA of CDglyTK-expressing Hep-2 cells by RT-PCR and a fusion protein of 59 000 was detected in cell extract from transfected Hep-2 cells. In vitro study growth of CDglyTK-positive Hep-2 cells were inhibited by 5-FC, GCV or 5-FC + GCV respectively, and the antitumour effect of 5-FC + GCV is superior to 5-FC or GCV.
  
  
  CONCLUSIONS
  CDglyTK may be a candidate for treating human laryngeal cancer.
authors.affiliation: Department of Otolaryngology, Hospital of Xiangya, Central South University, Changsha 410008, China.
authors.forename: Yaoyun
authors.initials: Y
authors.lastname: Tang
authors.forename: Changning
authors.initials: C
authors.lastname: Xie
authors.forename: Jianping
authors.initials: J
authors.lastname: Liu
authors.forename: Suping
authors.initials: S
authors.lastname: Zhao
authors.forename: Yongquan
authors.initials: Y
authors.lastname: Tian
authors.forename: Jianyun
authors.initials: J
authors.lastname: Xiao
mesh_terms: D045744:Cell Line, Tumor; D043525:Cytosine Deaminase / Q000235:genetics; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007822:Laryngeal Neoplasms / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics