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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)安全而可靠的治疗手段之一
Objective To investigate the differentiation of HL 60 cells induced by low-dose arabin C (Ara C) and its possible mechanism by using the characteristics of HL 60 cell differentiation. Methods (1) HL 60 cells were collected at 0, 2, 4, 6, 8 days after Ara C 10ng/ml treatment, and the expression of CD13, CD14, CD15, and CD11b was detected by flow cytometry (FCM). Cell cycle analysis; (2) semi-quantitative reverse transcription polymerase chain reaction (RT PCR) was used to detect the expression of bcl 2 at different time points; (3) cell smears were stained with Wright’s staining and cell morphology was observed under light microscope. Results (1) With the prolongation of drug action time, cells in S phase gradually decreased, while cells in G0/G1 phase gradually increased, and proliferation of HL 60 cells was inhibited; (2) HL 60 cells were partially transferred after a certain period of drug action. Nuclear and partial differentiation into granulocytes; (3) Expression of anti-apoptosis factor bcl 2 at the mRNA level decreased in a time-dependent manner; (4) Characteristics of HL 60 cell morphology gradually differentiated or apoptotic under the action of drugs . Conclusion After induction of low-dose Ara C in HL 60 cells, both the mononuclear and granulocyte differentiations were observed, and with the prolonged action time of low-dose Ara C, the apoptosis or cytotoxicity may be more pronounced. It is one of the safe and reliable treatments for clinical acute nonlymphocytic leukemia (ANLL)