OBJECTIVETo construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.

METHODSThe 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.

CONCLUSIONKH gene may be a leukemia gene related to proliferation of K562 cells.

Animals ; COS Cells ; Cell Proliferation ; Cercopithecus aethiops ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; physiology ; Genetic Vectors ; Humans ; K562 Cells ; L-Lactate Dehydrogenase ; metabolism ; Open Reading Frames ; genetics ; physiology ; Plasmids ; RNA, Messenger ; biosynthesis ; genetics ; RNA-Binding Proteins ; genetics ; S Phase ; Transfection