氨氯吡咪诱导白血病K-562细胞酸化及凋亡研究

目的研究氨氯吡咪诱导白血病细胞株K-562细胞酸化及凋亡的作用。方法采用荧光探针法检测氨氯吡咪对人白血病细胞株K-562细胞的酸化作用;采用流式细胞仪观察氨氯吡咪对K-562细胞周期的影响及诱导细胞凋亡的情况。结果与0 h相比,100μmol·L~(-1)氨氯吡咪处理8 h即能明显酸化K-562细胞,细胞内pH明显降低(P<0.05)。与0μmol·L~(-1)比较,除50μmol·L~(-1)氨氯吡咪处理K-562细胞24 h后G_1期细胞增加差异无统计学意义(P>0.05)外,其余各浓度氨氯吡咪处理K-562细胞24 h后G_1期细胞增加(P<0.01),S期细胞降低(P<0.01),G_2/M期细胞也有所增加(P<0.05)。与0μmol·L~(-1)相比,其余各浓度氨氯吡咪对K-562细胞处理24 h后细胞凋亡率明显增加(P<0.01)。与0 h相比,100μmol·L~(-1)氨氯吡咪对K-562细胞处理8、24、48 h凋亡细胞比率均明显增加(P<0.01)。结论氨氯吡咪可有效酸化K.562细胞,使细胞内pH显著降低,并诱导K-562细胞凋亡,肿瘤细胞的增殖生长也显著受到抑制,在白血病治疗中可能具有一定的应用前景。

Induction of the acidification and apoptosis of leukemia K-562 cell with amiloride

Objective To study the acidification and apoptosis of leukemia K-562 cell induced by amiloride.Methods The acidification of human leukemia K-562 cell induced by amiloride was determined with fluorescence probe method;Influence of cell cycle and induction of cell apoptosis was determined with flow cytometry.Results Compared with 0 hour,K-562 cells was acidated obviously if handled in 100μmol·L~(-1) amiloride for more than 8 hours,and the pHi was significantly lower(P<0.05 ).Compared with 0μmol·L~(-1),there was no statistically differences in the cell increases during G_1 phase when K-562 cell was handled in 50μmol·L~(-1) amiloride for 24 hours(P>0.05);Besides,when K-562 cell was handled in amiloride with other concentrations for 24 hours,cells in increased during G_1 phase(P<0.01 ),cells reduced during S period(P<0.01 ),cells increased during G_2/M(P<0.05).Compared with 0μmol·L~(-1),the apoptosis rate induced by K-562 cell handled in other concentrations of amiloride after 24 hours increased obviously(P<0.01 ).Compared with the 0 hour,there were significant increase in apoptosis rate induced by 100μmol·L~(-1) amiloride at different time(P<0.01 ).Conclusion Amiloride can induced the acidification and apoptosis of K-562 cells,and the proliferation and the growth of cancer cell were inhibited.It probably has a practical prospect in the treatment of leukemic.