title: [Construction of a recombinant lentiviral vector for VHL and VHL shRNA and its effect on proliferation and apoptosis of renal cell carcinoma cells].
abstract: OBJECTIVE
  To construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.
  
  
  METHODS
  Lentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.
  
  
  RESULTS
  The recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).
  
  
  CONCLUSIONS
  VHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.
authors.affiliation: Department of Urology/State Key Laboratory of Kidney Diseases, General Hospital of PLA, Chinese PLA Medical School, Beijing 100853, China.E-mail: dannyshen881015@qq.com.
authors.forename: Donglai
authors.initials: D
authors.lastname: Shen
authors.forename: Xin
authors.initials: X
authors.lastname: Ma
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Gao
authors.forename: Xingtao
authors.initials: X
authors.lastname: Li
authors.forename: Liangyou
authors.initials: L
authors.lastname: Gu
authors.forename: Huijie
authors.initials: H
authors.lastname: Gong
authors.forename: Shaoxi
authors.initials: S
authors.lastname: Niu
authors.forename: Xu
authors.initials: X
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis; D002292:Carcinoma, Renal Cell / Q000473:pathology*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D010957:Plasmids; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014162:Transfection; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human