title: [Antisense hTERT inhibits gene expression and functional activity of telomerase in leukemia cell lines].
abstract: OBJECTIVE
  To explore the inhibitory effects of gene expression and functional activity of telomerase in leukemia cell lines by in vitro antisense hTERT treatment.
  
  
  METHODS
  An antisense hTERT eukaryotic expression vector was constructed by using gene recombination technique, targeting the 5' end mRNA sequence of the telomerase catalytic subunit. The vector expression in leukemia cell lines (HL60 and K562) was achieved by transfection using the SuperFect transfection reagent (Qiagen). After transfection, ectopic expression of the telomerase catalytic subunit was analyzed by quantitative fluorescence real-time RT-PCR, and cellular apoptosis and cell cycle parameters were evaluated by flow cytometry respectively.
  
  
  RESULTS
  An antisense pcDNA-hTERT eukaryotic expression vector was successfully constructed. Leukemia cell lines transfected with antisense hTERT constructed displayed a significant inhibition of gene expression of telomerase and its activity in vitro, as compared with the result of the control groups (without transfection and vector control).
  
  
  CONCLUSIONS
  In-vitro antisense hTERT expression may down-regulate the gene expression and biological activity of telomerase in leukemia cells, suggesting a possibility of gene therapy against human malignancy through the telomerase-targeted molecular mechanism.
authors.affiliation: Department of Pathology, Shenzhen Children's Hospital, Shenzhen 518026, China. sunlb163@hotmail.com
authors.forename: Lai-bao
authors.initials: LB
authors.lastname: Sun
authors.forename: Cheng-rong
authors.initials: CR
authors.lastname: Li
authors.forename: Jian-ming
authors.initials: JM
authors.lastname: Wen
authors.forename: Guo-bing
authors.initials: GB
authors.lastname: Wang
authors.forename: Meng
authors.initials: M
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
authors.forename: Ruo-xin
authors.initials: RX
authors.lastname: Li
mesh_terms: D017209:Apoptosis; D002453:Cell Cycle; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D015536:Down-Regulation; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D016372:RNA, Antisense / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D011994:Recombinant Proteins; D019098:Telomerase