Effect of microRNA-193b on C-KIT protein expression and biological behaviors of K562 cells.
- Author:
Xiao-Ning GAO
1
;
Ji LIN
;
Li GAO
;
Yi DING
;
Jing-Xin LI
;
Li-Li WANG
;
Li YU
Author Information
1. Department of Hematology, Chinese PLA General Hospital, Beijing, China.
- Publication Type:Journal Article
- MeSH:
Cell Differentiation;
genetics;
Cell Proliferation;
Humans;
K562 Cells;
MicroRNAs;
genetics;
Proto-Oncogene Proteins c-kit;
genetics;
metabolism;
Transfection
- From:
Journal of Experimental Hematology
2011;19(6):1343-1347
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this study was to investigate the effect of microRNA-193b (miR-193b) on C-KIT protein expression and biological behaviors in K562 cells. The FAM-labeled miR-193b mimic and negative control were respectively transfected into K562 cells using HiPerFect transfection reagent. The percentage of FAM-positive cells was monitored by flow cytometry. The levels of C-KIT and phosphorylated C-KIT protein were detected by Western blot. The cell growth was measured by CCK-8 reagent. The apoptosis of cells were analyzed by flow cytometry with Annexin V staining. The differentiation of cells were analyzed by flow cytometry with anti-CD11b or anti-CD15 staining. The results demonstrated that the percentage of FAM-positive cells was about 80% in miR-193b or negative control-transfected K562 cells. Compared with the negative control group, overexpression of miR-193b in K562 cells significantly inhibited the levels of C-KIT and phosphorylated C-KIT protein. Meanwhile, the cell growth was inhibited and the percentages of apoptotic cells, CD11b- or CD15-positive cells increased. It is concluded that miR-193b can reduce C-KIT expression and inhibit cell growth in K562 cells. The growth-inhibitory activity of miR-193b is associated with apoptosis and granulocytic differentiation. This study contributed to further investigate the role of miR-193b in leukemogenesis.
