title: [Construction of recombinant lentiviral vectors containing Rheb gene and its mutant Rheb'D60K gene and their expression in human liver cancer cells].
abstract: OBJECTIVE
  To construct recombinant lentiviral vectors carrying Rheb gene and its mutant Rheb'D60K gene, and examine their expression in human liver cancer cells.
  
  
  METHODS
  Rheb gene was amplified by PCR to construct the recombinant plasmid LV31-Rheb-WT and LV31-Rheb-D60K. HEK-293 FT cells were contransfected with the recombinant lentiviral vector together with a lentiviral package plasmid to produce the lentiviral particles. The expression of PS6 protein was detected in the lentivirus-infected MCF-7 cells. The apoptosis of SK-HEP-1 cells transfected with LV31-Rheb-WT or LV31-Rheb-D60K was observed.
  
  
  RESULTS
  The recombinant LV31-Rheb-WT and LV31-Rheb-D60K vectors were confirmed by PCR and DNA sequencing. Western blotting showed that PS6 protein expression was increased in LV31-Rheb-WT-transfected cells while decreased in LV31-Rheb-D60K-transfected cells. LV31-Rheb-D60K-transfected SK-HEP-1 cells showed more obvious apoptosis after starvation than LV31-Rheb-WT-transfected cells.
  
  
  CONCLUSIONS
  Lentiviral vectors carrying Rheb gene and its mutant has been successfully constructed, which can be useful in further investigation of the role of Rheb gene in cancer cells.
authors.affiliation: Clinical Oncology Center, People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China. ckhxy@163.com
authors.forename: Ke-He
authors.initials: KH
authors.lastname: Chen
authors.forename: Bo
authors.initials: B
authors.lastname: Liang
authors.forename: Zhen-Hong
authors.initials: ZH
authors.lastname: Zou
authors.forename: Ze-Long
authors.initials: ZL
authors.lastname: Han
authors.forename: Jin-Fei
authors.initials: JF
authors.lastname: Pan
authors.forename: An-Ling
authors.initials: AL
authors.lastname: Liu
mesh_terms: D017209:Apoptosis / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology*; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D061986:MCF-7 Cells; D020559:Monomeric GTP-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics*; D050505:Mutant Proteins / Q000235:genetics*; D009479:Neuropeptides / Q000096:biosynthesis* / Q000235:genetics*; D000076205:Ras Homolog Enriched in Brain Protein; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D050505:Mutant Proteins; D009479:Neuropeptides; C490211:RHEB protein, human; D000076205:Ras Homolog Enriched in Brain Protein; D011994:Recombinant Proteins; D020559:Monomeric GTP-Binding Proteins