title: [Construction of eukaryotic expression vector of unknown KH gene and its effect on cell proliferation].
abstract: OBJECTIVE
  To construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.
  
  
  METHODS
  The pCI-neo-KH-ORF expression vector was constructed by DNA recombinant technique and was introduced into COS-7 cells and K562 cells by lipofectactin-mediated DNA transfection. Expression of KH-ORF mRNA was detected by RT-PCR. The effect of KH-ORF on cell cycle of COS-7 cells and K562 cells was evaluated by flow cytometry (FCM). Effect on cell proliferation of COS-7 cells was tested by MTT assay and that on K562 cells was analyzed by growth curves and LDH activity measurement.
  
  
  RESULTS
  (1) KH-ORF mRNA was expressed both in COS-7 cells and K562 cells. (2) The cell cycle and cell proliferation of COS-7 cells were unaffected significantly. (3) The proportion of cells in S phase was increased in pCI-neo-KH-ORF-transfected K562 cells; and growth curves and LDH activity indicated enhanced cell proliferation.
  
  
  CONCLUSIONS
  KH gene may be a leukemia gene related to proliferation of K562 cells.
authors.affiliation: Department of Clinical Biochemistry, Chongqing University of Medical Sciences, Chongqing 400016, China.
authors.forename: Yu-juan
authors.initials: YJ
authors.lastname: He
authors.forename: Yi-heng
authors.initials: YH
authors.lastname: Ou
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xiao-shan
authors.initials: XS
authors.lastname: Liu
authors.forename: Ling-di
authors.initials: LD
authors.lastname: Ma
authors.forename: Ji-kai
authors.initials: JK
authors.lastname: Jiang
mesh_terms: D000818:Animals; D019556:COS Cells; D049109:Cell Proliferation; D002522:Chlorocebus aethiops; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D020014:K562 Cells; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics* / Q000502:physiology; D010957:Plasmids; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics; D016196:S Phase; D014162:Transfection
chemical_list: D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D007770:L-Lactate Dehydrogenase