Effect of microRNA-193b on C-KIT protein expression and biological behaviors of K562 cells / 中国实验血液学杂志

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.