As_2O_3诱导白血病细胞凋亡过程中Ca^(2+)的变化及其调节

Changes of Ca^(2+) and its regulation in apoptosis of leukemia cells induced by As_2O_3

目的 研究Ca2 +在As2 O3 诱导白血病细胞系NB4、K5 6 2、HL 6 0凋亡中的作用 ,及抗氧化剂NAC、NDMS、CAT和Ca2 +清除剂Quin 2对As2 O3 引起的Ca2 +变化的调控作用。方法 细胞内Ca2 +用Fluo 3AM标记 ,并用流式细胞仪检测。结果 0 .6 μmol/LAs2 O3 作用 72h能诱导NB4发生 4 4 .9%的凋亡 ,提高其浓度至 2 .7和 8.1μmol/L ,也能诱导K5 6 2和HL 6 0分别发生 5 8.3%和 6 2 .3%凋亡。As2 O3 能不同程度降低NB4和HL 6 0细胞内Ca2 +水平 ,对K5 6 2细胞内Ca2 +水平影响不大。抗氧化剂NAC、NDMS、CAT和Ca2 +清除剂Quin 2对As2 O3 引起的NB4细胞Ca2 +水平下降显示出抑制作用 ,对HL 6 0细胞内Ca2 +水平下降无明显抑制作用。结论 Ca2 +水平下降在As2 O3 诱导的不同白血病细胞凋亡作用过程起着不同的作用。抗氧化剂NAC、NDMS、CAT和Ca2 +清除剂Quin 2可不同程度地抑制As2 O3 诱导的NB4细胞凋亡时细胞内Ca2 +水平下降。

Objective To investigate the effect of arsenic trioxide (As 2 O 3 ) on Ca 2+ levels in 3 leukemia cell lines NB4, K562 and HL-60. Regulation effects of antioxidants, thiol reagents (N-acetyl-l-cysteine, NAC; Natrii dimercaptosussinas, NDMS), catalase (CAT) and calcium chelator (Quin2) on Ca 2+ levels were also examined during these processes. Methods Ca 2+ were labeled with Fluo-3 AM and measured with flow cytometry(FCM). Results The results showed that As 2 O 3 (0.6 μmol/L) induced 44.9 % apoptosis in NB4 cells only, however, when concentration of As 2 O 3 increased to 2.7 and 8.1 μmol/L respectively, 58.3 % and 62.3 % apoptosis significantly exhibited in K562 and HL-60 cells. At the above 3 concentrations, As 2 O 3 decreased Ca 2+ markedly in NB4 and HL-60 cells, but not in K562 cells. Antioxidants NAC, CAT, NDMS, and calcium chelator Quin 2 showed inhibition effects on the decline of Ca 2+ levels only in NB4 cells. Conclusion The declines of intracellular Ca 2+ levels could play various roles in different leukemia cells in apoptosis induced by As 2 O 3 . Antioxidants and calcium chelator are able to inhibit the declines of intracellular Ca 2+ in NB4 cells during apoptosis induced by As 2 O 3 .