阿糖胞苷体外对人白血病细胞株HL-60细胞增殖抑制和诱导凋亡的相关性研究

Study on the proliferation inhibition and apoptosis induction of Ara-C on treatment HL-60 in vitro

目的 研究阿糖胞苷 (Ara -C)在体外对人白血病细胞株HL - 6 0增殖抑制和诱导凋亡的剂量 -时间 -效应关系。通过对Ara -C在不同剂量和疗程时抗白血病的机理研究 ,验证大剂量阿糖胞苷 (HD -AraC)治疗血液肿瘤的优越性。方法 在体外设定的浓度梯度和作用时间下应用Ara -C处理人白血病HL - 6 0细胞 ,采用四氮唑盐还原法 (MTT法 )检测Ara -C对HL - 6 0细胞增殖抑制的剂量 -时间 -效应关系 ,荧光染色观察凋亡细胞形态学改变 ,琼脂糖凝胶电泳测定DNA梯带 (DNA ,Lad der) ,流式细胞仪检测细胞凋亡率和细胞周期变化。结果 荧光染色观察到特异性的凋亡小体形成 ,DNA琼脂糖凝胶电泳显示出典型的电泳条带变化 ,流式细胞仪检查见明显的凋亡峰形成 ,细胞周期主要阻滞在sub -G1期。MTT法显示阿糖胞苷作用于HL - 6 0细胞的半数抑制浓度 (IC50 )分别是 2 .4 1 μM和 1 1 .81 μM。统计分析表明 :随着药物浓度的增加和作用时间的延长 ,Ara -C对细胞的增殖抑制和诱发凋亡作用逐渐增强 (P <0 .0 5)。结论 诱导白血病细胞凋亡可能是Ara -C抗白血病的机理之一 ,通过诱导HL - 6 0细胞凋亡从而对其产生明显的增殖抑制作用 ,且与剂量、时间相关。提示应用HD -AraC治疗白血病 ,可以明显提高疗效。

Objective To investigate the relationship of dose-duration-effect of proliferation inhibition and apoptosis induction when treated HL-60 with different dose of Ara-C and different duration of administration in vitro. In addition to study the antileukemic mechanism of Ara-C. Methods Following treat the HL-60 cell with designed concentration and duration, the MTT survey was used to detect the relationship of dose-duration-effect in inhibiting the proliferation of HL-60. Then the morphology of HL-60 and the DNA Ladder were observed through luminescent dyes-Hoechst 33258 and agar electrophoresis respectively. Furthermore, the cell cycle distribution and apoptosis rate were determined through FCM. Results The apoptosis body and DNA Ladder were observed apparently and the marked apoptotic peak were inspected, and the cell cycle were retarded in sub-G1. In the MTT survey, the IC50 were 11.81 μM and 2.41 μM in one-day group and two-day group respectively. The statistics results showed that the proliferation inhibition rate and apoptosis rate were increased corresponding to the improve of concentration and the prolong of treatment duration (P<0.05). Conclusion The antileukemic mechanism of Ara-C might be induce leukemia cells to generate apoptosis, thereby cause the noticeable inhibition of proliferation. The results suggests that the outcomes of therapy for leukemia could be significantly elevated by using high-dose Ara-C.