三氧化二砷对慢性粒细胞白血病细胞系的作用

摘要

The objective is to explore the effect and the mechanism of arsenic trioxide (As2O3) on different cell lines of chronic myeloid leukemia (CML). Different concentrations of As2O3 (0.2, 2 and 10μmol/L) were added to CMl cell lines KU812 and MEG-01 and other leukemia cell lines U937 and Pl21, the cell numbers were counted at different times, TUNEL and DNA ladder were assayed. Different antibodies, CD34, CD13, CD33,CD19.CD11b,CD14 and CD7, were added to detect the change of the molecules on cell surface, the change of bcr-abl ty RT-PCR and the activity of caspase-3 were assayed. The results showed that different concentrations of As2O3 had different effects on the sruvival of the 4 cell lines. After culture for 24 hours with As2O3, there was no significant increase in CD11b in all the four cell lines. There were no changes of bcr-abl in the 2 CML cell lines treated and untreated with As2O3 by RT-PCR. Activities of caspase-3 were all increased. It is concluded that As2O3 can induce apoptosis in CMl cell lines, the concentration to induce apoptosis is different, CML cell lines are more sensitive than the other 2 leukemia cell lines. As2O3 induced apoptosis may have some relation with the activation of capases-3.%本研究探讨三氧化二砷(As2O3)对两种慢性粒细胞白血病(CML)细胞系有无作用及作用机制.实验选用了两种不同的CML细胞系KU812和MEG-01及两种其它白血病细胞系U937和PL21,加入不同浓度的As2O3(0.2,2和10μmol/L),在不同时间计数活细胞数,观察细胞形态,并以TUNEL法和DNA凝胶电泳检测细胞有无凋亡的发生;同时检测0.2 μmol/L As2O3作用时细胞表面CD34,CD13,CD33,CD19,CD11b,CD14和CD7的变化,并通过RT-PCR检测细胞bcr-abl水平及caspase-3的变化.研究结果发现,不同浓度的As2O3对4种细胞系的作用不同,10μmol/L时4种细胞系均会发生细胞凋亡,2μmol/L时则仅CML细胞系KU812和MEG-01会产生凋亡,0.2μmol/L时4种细胞系均不发生凋亡.在培养24小时后,4种细胞系表面的CD34,CD13,CD33,CD19,CD11b,CD14和CD7等均无明显改变.RT-PCR检测bcr-abl的水平发现在2种CML细胞系中无明显改变,其caspase-3的活性较对照细胞均有所增加.结论:As2O3在较高浓度下可诱导4种白血病细胞系产生凋亡,在2μmol/L仅可诱导CML来源的两种细胞系产生凋亡,CML细胞系对As2O3较其它两种细胞系敏感,其作用机制与caspase-3有关.