title: [Establishment of a pulmonary metastasis model of melanoma   with stably expressed hTERT/Luc].
abstract: OBJECTIVE
  To construct the eukaryotic expression vectors of human telomerase reverse transcriptase (hTERT) and luciferase (Luc) gene, and stably transfect B16 cell line with them, and inoculate mice via caudal vein to construct pulmonary metastasis models.
  
  
  METHODS
  The hTERT and Luc cDNA fragments were inserted into eukaryotic expression vectors to obtain the recombinant plasmids pIRES-neo-hTERT and pIRES-hyg3-Luc. The plasmids were then transfected into B16 cells by Lipofectamine(TM); 2000. Afte the stably transfected cell line was screened by G418 and hygromycin B, the transcription and expression of the hTERT and Luc genes were identified by Western blotting and immunofluorescence staining. The stably transfected cells were inoculated via caudal vein into male C56BL/6 mice to construct pulmonary metastasis models. An in vivo imaging system was used to monitor the growth and metastasis of the tumors.
  
  
  RESULTS
  A stably transfected cell line was established and the express rates of hTERT and Luc gene in the stable cell line were 84% and 98%, respectively. Pulmonary metastasis models were successfully established and the conditions of the lung metastasis could be detected easily using in vivo imaging system.
  
  
  CONCLUSIONS
  The pulmonary metastasis models with stably expressed hTERT for in vivo imaging system in mice has been successfully established.
authors.affiliation: Department of Urology, Chinese PLA General Hospital, Beijing 100853, China. xiaoyi1007@gmail.com
authors.forename: Yi
authors.initials: Y
authors.lastname: Xiao
authors.forename: Jiangping
authors.initials: J
authors.lastname: Gao
authors.forename: Kun
authors.initials: K
authors.lastname: Gao
authors.forename: Jinqi
authors.initials: J
authors.lastname: Yan
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.forename: Yuanji
authors.initials: Y
authors.lastname: Xu
authors.forename: Xiaoxiong
authors.initials: X
authors.lastname: Wang
authors.forename: Jiyun
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D015870:Gene Expression; D006801:Humans; D008156:Luciferases / Q000235:genetics*; D008175:Lung Neoplasms / Q000235:genetics* / Q000556:secondary*; D008297:Male; D008546:Melanoma, Experimental / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D009362:Neoplasm Metastasis; D010957:Plasmids / Q000235:genetics; D019098:Telomerase / Q000235:genetics*
chemical_list: D008156:Luciferases; C509186:TERT protein, human; D019098:Telomerase