Construction of eukaryotic expression vector of unknown KH gene and its effect on cell proliferation.
- Author:
Yu-juan HE
1
;
Yi-heng OU
;
Yan ZHANG
;
Xiao-shan LIU
;
Ling-di MA
;
Ji-kai JIANG
Author Information
- Publication Type:Journal Article
- MeSH: 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
- From: Chinese Journal of Oncology 2005;27(3):129-133
- CountryChina
- Language:Chinese
-
Abstract:
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.
