MicroRNA-10b通过调控锌指蛋白KLF4阻滞白血病细胞分化  

MicroRNA-10b Blocks the Differentiation of Leukemia Cells Through Regulating the Zinc Finger Protein KLF4

作　　者：谭诗[1] 张慧娟[1] 王娟[1] 陈莎娜[1] 全静[1] 鲜敬荣[1] 张伶[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室重庆市重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2013年第3期290-295,共6页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81271913)资助的课题~~

摘　　要：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。We aimed to explore the effect of microRNA-10b （miR-10b） on the differentiation of leukemia cells through regulating the expression of zinc finger protein Kriippel-like factor 4 （KLF4）. The expression of miR- 10b and KLF4 in leukemia cell lines at different levels of differentiation was detected by Real-time PCR and Western blot, respectively. Leukemia cell line HL60 was induced with 1,25-dihydroxy-vitamin D3 （1,25D3） to differentiate along the monocytic lineage. The expression of miR-10b and KLF4 was examined during 1,25D3-induced differentiation of HL60. The synthesized miR-10b mimics was transfected into HL60 cells. The morphological changes of cells treated with 1,25D3 were observed under light microscope following Wright-Giemsa staining, and the monocyte surface marker CD14 was analyzed by flow cytometry, miR-10b was detected at the highest levels of expression in KG-la cells displaying early differentiation phenotypes and at the lowest in more mature U937 and THP-1 cells （P〈0.01）,while the KLF4 exhibited an opposite expression pattern, miR-10b was decreased in a time-dependent manner in HL60 cells during induction with 1,25D3, whereas the KLF4 was increased. Enforced expression ofmiR-10b in HL60 cells inhibited the 1,25D3-induced morphological changes and the expression of CD14 （P〈0.05）. Our data indicate that miR-10b suppresses monocytic differentiation of ilL60 cells via targeting to KLF4.

关 键 词：MIR-10B KLF4 急性髓系白血病 细胞分化 

分 类 号：R733.7[医药卫生—肿瘤]