【摘要】 目的 探讨中药砒霜的主要有效成分三氧化二砷（As₂O₃）对白血病干细胞（LSCs）的增殖、凋亡和自我更新能力的影响。方法 以药物敏感细胞株K562细胞和白血病多药耐药株K562/ADM细胞为靶细胞，利用免疫磁珠法分选纯化K562细胞和K562/ADM细胞中的LSCs;MTT比色法测定As₂O₃对细胞增殖活性的影响；流式细胞术结合AnnexinⅤ/PI染色法测定细胞凋亡率；流式细胞术结合LSCs细胞表面标志物检测细胞群体中LSCs相对含量；甲基纤维素集落形成法检测细胞的自我更新和增殖能力。结果 低浓度As₂O₃作用后，K562细胞和K562/ADM细胞中LSCs的含量不同程度增高，高浓度时增高较缓或不明显，但凋亡LSCs的数量则随浓度的增高而增高；As₂O₃呈浓度依赖性地抑制两类LSCs（敏感性LSCs和耐药性LSCs）的增殖，并诱导部分LSCs凋亡，多药耐药性LSCs对As₂O₃的敏感性较强；As₂O₃显著抑制敏感性LSCs和耐药性LSCs的集落形成能力，且对耐药性LSCs的抑制效应更强。结论 As₂O₃能够抑制LSCs的增殖并诱导其凋亡，来源于耐药性白血病细胞群体的多药耐药性LSCs对As₂O₃的敏感性更高。更多还原

【Abstract】 Objective To investigate the inhibitory and apoptotic effects of arsenic trioxide （As₂O₃） on leukemia stem cells （LSCs）.Methods K562/ADM cell line,a multidrug-resistant leukemia cell line and its drug-sensitive parent cell line K562 were used as cell models.LSCs were purified via MiniMACS CD34,CD38 and CD123 isolation kits following the manufacture’s protocol.Cell proliferation was measured by MTT assay,and cell apoptosis was measured using AnnexinⅤ/PI staining.The relative content of LSCs was detected by flow cytometry,and the self-renewal and proliferation ability of cells detected by colony formation assay.Results The purity and viability of LSCs sorted from K562 and K562/ADM cell populations were both above 90%.The content of LSCs in K562 and K562/ADM cells increased by varying degrees at low concentrations,but increased slowly or not at all at high concentrations.However,the number of apoptotic LSCs increased with increasing concentration.As₂O₃ inhibited the proliferation of both types of LSCs (LSC_K562 and LSC_K562/ADM) in a concentration-dependent manner and induced apoptosis in some LSCs.Drug-resistant LSCs were more sensitive to As₂O₃.As₂O₃ significantly inhibited the formation of LSC_K562and LSC_K562/ADM colonies on semi-solid culture medium,and the inhibitory effect of As₂O₃ on LSC_K562/ADM was stronger.Conclusion As₂O₃ can inhibit the proliferation of LSCs and induce apoptosis.LSCs derived from drug-resistant leukemia cell populations are more sensitive to As₂O₃.更多还原