Construction of eukaryotic expression vector of unknown KH gene and its effect on cell proliferation / 中华肿瘤杂志
Chinese Journal of Oncology
;
(12): 129-133, 2005.
Article
Dans Chinois
| WPRIM
| ID: wpr-331210
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic expression vector of open reading frame of unknown KH gene (KH-ORF), and investigate its effect on cell proliferation.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>(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.</p><p><b>CONCLUSION</b>KH gene may be a leukemia gene related to proliferation of K562 cells.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Physiologie
/
Plasmides
/
ARN messager
/
Transfection
/
Régulation de l'expression des gènes tumoraux
/
Chlorocebus aethiops
/
Cadres ouverts de lecture
/
Phase S
/
Protéines de liaison à l'ARN
/
Cellules COS
Limites du sujet:
Animaux
/
Humains
langue:
Chinois
Texte intégral:
Chinese Journal of Oncology
Année:
2005
Type:
Article
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