K562细胞硫氧还蛋白还原酶活力测定及其抑制剂体外抗白血病的初步研究

Preliminary study on the thioredoxin reductase in K562 cells and anti-leukemia effect of BBSKE invitro

目的探讨慢性粒细胞白血病（CML）细胞株K562中硫氧还蛋白还原酶（TrxR）的活力及其新型抑制剂乙烷硒啉（BBSKE）体外抗白血病作用。方法应用胰岛素还原法检测K562细胞株及健康人骨髓单个核细胞中TrxR的活力。运用CCK-8法测定BBSKE对K562细胞的增殖抑制率。应用激光共聚焦显微镜、琼脂糖凝胶电泳以及AnnexinV-FITC／PI双标记流式细胞术观察BBSKE的抗白血病作用。结果K562细胞中TrxR的活性明显高于健康人骨髓单个核细胞，10Ixmol／LBBSKE与K562细胞作用24h，激光共聚焦显微镜可见典型的细胞凋亡表现，琼脂糖凝胶电泳后可见典型的DNA“梯”条带出现，流式细胞术检测凋亡率为（10．28±2．74）％；10μmol／LBBSKE对CML患者原代细胞有诱导凋亡的作用，凋亡率为（5．70±0．48）％。结论慢性粒细胞白血病细胞株K562中TrxR活力高于健康人骨髓单个核细胞，BBSKE有抑制TrxR活力、抑制K562细胞增殖和诱导凋亡的作用，是治疗CML潜在的有效药物。

Objective To explore the activity of thioredoxin reductase （TrxR） in chronic myeloid leukemia cell line K562 and the anti-leukemia effect of BBSKE （a novel inhibitor of TrxR） in vitro. Methods The activity of TrxR on K562 cell lineage and fresh bone marrow cell from healthy adult was analyzed by insulin reduction assay. The inhibition of proliferation was measured by CCK-8 assay. The anti-leukemia effect of BBSKE was detected by laser scanning confocal microscope,agarose gel electrophoresis and flow cytometry with Annexin V-FITC/PI staining, Results TrxR activity of K562 cell lineage was significantly higher than that of normal bone marrow mononuclear ceils. The apoptosis of K562 cells could be induced at concentrations of 10 μmol/L BBSKE after treated for 24 hours. The typical DNA ＂ladder＂ bans were observed by agarose gel electrophoresis. The apoptotie rates of K562 cells were （10.28±2.74） %. Application of 10 p, mol/L BBSKE for 48 hours could also induce apoptosis of fresh bone marrow cell from chronic myeloid leukemia patients, and the apoptotic rates were （5.70±0.48） %. Conclusion TrxR activity in chronic myeloid leukemia cells was significantly higher than that of normal cells. BBSKE inhibits the TrxR activity and the proliferation of K562 by inducing apoptosis.It might be a potential medication for chronic myeloid leukemia.