论文部分内容阅读
探讨microRNA-10b(miR-10b)通过调节锌指蛋白Krüppel-like factor 4(KLF4)的表达对急性白血病细胞分化的影响。Real-time PCR及Western blot分别检测不同分化程度的白血病细胞系中miR-10b与KLF4的表达;1,25-二羟基维生素D3(1,25D3)诱导人白血病细胞系HL60向单核系分化,检测此过程中miR-10b及KLF4的表达变化;利用体外合成的寡核苷酸(miR-10b mimics)转染HL60细胞,瑞氏–吉姆萨染色观察1,25D3诱导后细胞分化形态学的改变;流式细胞术检测单核细胞表面标志CD14的表达。结果显示,miR-10b在分化早期的KG-1a细胞中表达最高,在分化晚期的U937、THP-1细胞中表达最低(P<0.01),而KLF4的表达与之相反;1,25D3诱导HL60向单核系分化过程中,miR-10b表达呈时间依赖性降低,KLF4表达则逐渐增高;HL60细胞中过表达miR-10b后可抑制1,25D3诱导的细胞分化形态特征的改变及CD14的表达(P<0.05)。提示miR-10b通过负调控KLF4的表达阻滞白血病细胞HL60单核系的分化。
To investigate the effect of microRNA-10b on the differentiation of acute leukemia cells by regulating the expression of KLF4. Real-time PCR and Western blot were used to detect the expression of miR-10b and KLF4 in different differentiated leukemia cell lines; 1,25-dihydroxyvitamin D3 (1,25D3) induced differentiation of human leukemia cell line HL60 to monocytogenes, The expression of miR-10b and KLF4 in this process was detected. HL60 cells were transfected with miR-10b mimics in vitro. Wright-Giemsa staining was used to observe the morphological changes of cells after 1,25D3 induction Flow cytometry was used to detect the expression of monocyte surface marker CD14. The results showed that miR-10b was the highest expression in early differentiation of KG-1a cells, the lowest expression in U937 and THP-1 cells in late differentiation (P <0.01), while the expression of KLF4 was the opposite; In the process of monocyte differentiation, the expression of miR-10b decreased in a time-dependent manner and the expression of KLF4 gradually increased. The overexpression of miR-10b in HL60 cells inhibited 1,25D3-induced morphological changes of cell differentiation and CD14 expression (P <0.05). It is suggested that miR-10b blocks the differentiation of HL60 monocytogenes by negatively regulating KLF4 expression.