去甲斑蝥素对白血病细胞系K562的增殖、凋亡、细胞周期及NF-κB活性的影响  

Effect of norcantharidin on the proliferation,apoptosis,cell cycle and activity of NF-κB on human leukemia cell line K562

作　　者：姚铠涛[1] 陈江声[1] 陈春莉[1] 

机构地区：[1]汕头大学医学院第二附属医院血液肿瘤科,广东汕头515041

出　　处：《临床肿瘤学杂志》2015年第5期406-410,共5页Chinese Clinical Oncology

摘　　要：目的：探讨去甲斑蝥素（ NCTD）对白血病细胞系K562的增殖、凋亡、细胞周期及核因子（ NF）-κB活性的影响。方法采用CCK-8试剂盒检测不同浓度NCTD（0、10、25、50、100μmol／L）处理K562细胞24、48、72、96 h后的增殖抑制率，采用流式细胞术检测不同浓度NCTD处理24、48 h后的细胞凋亡率和处理48 h后的细胞周期，采用原位细胞凋亡检测试剂盒（ Tunnel）检测不同浓度NCTD处理24、48 h后的细胞凋亡指数，采用Western blotting检测不同浓度NCTD处理48 h后的NF-κB p65及IκB-α水平以评价NF-κB活性的变化。结果在10～100μmol／L范围内，NCTD可呈时间和浓度依赖的方式升高K562细胞的增殖抑制率，各浓度间的差异有统计学意义（ P＜0.05）；与0μmol／L相比，其余各浓度处理24、48 h后的凋亡率和凋亡指数均升高，处理48 h后的G0／G1期细胞比例升高，S、G2／M期细胞比例均降低，NF-κB p65水平降低，IκB-α水平升高，以上差异均有统计学意义（ P＜0-05）。结论 NCTD对白血病细胞系K562有细胞毒性，可抑制该细胞的增殖、诱导其凋亡及细胞周期阻滞并下调NF-κB表达。Objective To investigate the effect of norcantharidin （ NCTD） on proliferation, apoptosis, cell cycle and the ac-tivity of nuclear factor （ NF）-κB on human leukemia cell line K562. Methods The K562 cell was treated with different concentrations of NCTD （ 0, 10, 25, 50, 100μmol/L） and the proliferation inhibition rates were measured by CCK-8 kit at 24, 48, 72, 96 h post-treatment. The apoptotic rates at 24 and 48 h together with cell cycle at 48 h after NCTD treatments were evaluated via flow cytometry. The in situ cell death detection kit （ Tunnel） was applied to measure the levels of apoptosis at 24 and 48 h post-treatment. As for the activity of NF-κB, levels of p65 subunit of NF-κB and IκB-α were detected by Western blotting at 48 h post-treatment. Results NCTD in the range 10 to 100μmol/L could increase the proliferation inhibition rates on K562 cells in a dose-and time-dependent man-ner （ P〈0.05） . In comparison with 0μmol/L NCTD, there were elevated apoptotic rates and indices at 24 and 48 h post-treatment （ P〈0.05）. The treatment with NCTD （10.100μmol/L） resulted in a higher proportion of cells within G0/G1phase but a lower proportion of cells within S and G2/M phases with significant differences than 0μmol/L （ P〈0.05） . Reduced IκB-αbut elevated p65 subunit of NF-κB were observed only in 10.100 μmol/L NCTD （ P〈0.05） . Conclusion NCTD exhibited cytotoxity on K562 cells, possibly by the inhibition of the cell proliferation, induction of apoptosis and down-regulation of the activity of NF-κB.

关 键 词：去甲斑蝥素 白血病 细胞增殖 凋亡 细胞周期 

分 类 号：R733.7[医药卫生—肿瘤]