小剂量阿糖胞苷体外诱导HL-60细胞定向分化的实验研究  

Study of directional differentiation of HL-60 cell by low-dose cytosine arabinside in vitro

作　　者：沈群[1] 王毅军[1] 夏雯[1] 傅行财[1] 候艳秋[1] 宋敏[1] 张柳波[1] 

机构地区：[1]南京医科大学第二附属医院血液科,江苏省中医院210011

出　　处：《江苏医药》2002年第10期735-737,共3页Jiangsu Medical Journal

基　　金：江苏省"3 3 3二期工程"项目

摘　　要：目的　利用HL 6 0细胞定向分化的特征 ,探讨小剂量阿糖胞甘 (Ara C)诱导HL 6 0细胞的分化方向及可能的作用机制。方法　 (1)分别于Ara C 10ng/ml处理后 0 ,2 ,4 ,6 ,8天收集HL 6 0细胞 ,采用流式细胞术 (FCM)检测CD13、CD14、CD15、CD11b的表达 ,并进行细胞周期分析 ;(2 )半定量逆转录 聚合酶链反应 (RT PCR)检测不同时段bcl 2的表达 ;(3)细胞涂片瑞氏染色 ,光镜下观察细胞形态变化。结果　 (1)随药物作用时间的延长 ,S期细胞逐渐减少 ,而G0 /G1期细胞逐渐增加 ,HL 6 0细胞增殖受抑 ;(2 )药物作用一定时间后 ,HL 6 0细胞部分向单核细胞、部分向粒细胞分化 ;(3)抗凋亡因子bcl 2在mRNA水平的表达呈时间依赖性下降 ;(4)药物作用下 ,HL 6 0细胞形态学渐出现分化或凋亡的特征。结论　HL 6 0细胞经小剂量Ara C诱导作用后 ,既有向单核 ,也有向粒细胞分化 ;小剂量Ara C随作用时间的延长 ,其细胞凋亡作用甚或细胞毒作用可能更为明显 ,不失为临床急性非淋巴细胞性白血病 (ANLL)Objective To discuss low dose cytosine arabinoside (Ara C)induced directional differentiation of HL 60 cells and the possible mechanism.Methods (1) Using FCM to analyze the changes of CD13,CD14,CD15,CD11b and cell cycles in HL 60 cells treated by Ara C(10ng/ml)at day 0,2,4,6,8.(2)Determining the expression of bcl 2 with semi quantity reverse transcript polymerase chain reaction (RT PCR) respectively.(3)Using Wrights staining to observe the morphological changes of cells with microscope.Results (1)In the course of induction,the G0/G1 phase cells increased in number but the S phase cells decreased,suggesting the proliferation of cells was suppressed.(2)Exposed HL 60 cells in low dose Ara C(10ng/ml)were directionally developed to granulocyte or monocyte like cells.(3)The expression of bcl 2 gene decreased in a time dependent manner.(4)The morphological changes showed cell differentiation or apoptosis characteristics.Conclusion HL 60 cells were induced to either granulocytic or monocytic lineage by handling low dose Ara C;It might be got more obviously in the presentation of apoptosis or cytotoxicity that treated with the continuous low dose Ara C in HL 60 in vitro.Therefore,we believe that low dose Ara C may be one of the safe and effective therapeutics in adult ANLL.

关 键 词：HL-60细胞 诱导分化 细胞凋亡 阿糖胞苷 流式细胞术 半定量逆转录-聚合酶链反应 急性非淋巴细胞性白血病 ANLL 

分 类 号：R733.71[医药卫生—肿瘤]