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目的:研究纳米雄黄对白血病K562细胞及其白血病干细胞的凋亡诱导作用。方法:采用机械研磨法制备纳米雄黄。以白血病K562细胞为靶细胞,MTT法检测K562细胞增殖活性,采用流式细胞术检测K562细胞凋亡率、P-gp和BCRP蛋白的表达水平,以及白血病干细胞含量变化及其凋亡。结果:雄黄经高能球磨机球磨10～50h,扫描电镜(SEM)和透射电镜(TEM)观察,电镜测量纳米雄黄的平均粒径为72.72±22.18 nm。20μg/ml和50μg/ml纳米雄黄及雄黄原药处理K562细胞后,纳米雄黄对K562细胞有明显的增殖抑制作用,24h、48h、72h的IC50值分别为43.48μg/ml、20.52μg/ml、16.07μg/ml。细胞的凋亡率明显升高。雄黄原药对K562细胞也有增殖抑制及凋亡诱导作用,但效果要差于纳米雄黄。BCRP、P-gp蛋白表达以及二者共表达率随着时间的延长和浓度的增高呈上升趋势,尤以高浓度时明显。20μg/ml和50μg/ml纳米雄黄处理K562细胞24h,LSC含量随浓度升高而升高,K562细胞群中LSC的凋亡率升高。结论:纳米雄黄可诱导药物敏感性白血病细胞及其白血病干细胞发生凋亡。
AIM: To investigate the apoptosis-inducing effect of nano-realgar on leukemia K562 cells and leukemia stem cells. Methods: Nano realgar was prepared by mechanical grinding method. The leukemia K562 cells were used as target cells. The proliferation of K562 cells was detected by MTT assay. The apoptosis rate of K562 cells, the expressions of P-gp and BCRP proteins, the changes of leukemia stem cells and apoptosis were detected by flow cytometry. Results: Realgar was ball-milled for 10-50h by high-energy ball mill, and observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The average diameter of realgar was 72.72 ± 22.18 nm. Nano-realgar could significantly inhibit the proliferation of K562 cells after 20μg / ml and 50μg / ml nano-realgar and realgar treatment. The IC50 values at 24h, 48h and 72h were 43.48μg / ml and 20.52μg / ml respectively, 16.07 μg / ml. Cell apoptosis rate was significantly increased. Realgar also inhibited the proliferation of K562 cells and induced apoptosis, but the effect was worse than that of nano-realgar. The expression of BCRP and P-gp protein and their co-expression rate increased with the increase of time and concentration, especially in high concentration. In 20μg / ml and 50μg / ml nano-realgar treatment K562 cells 24h, LSC content increased with increasing concentration, K562 cell population LSC apoptosis increased. Conclusion: Nano realgar can induce apoptosis of drug sensitive leukemic cells and leukemia stem cells.