目的 研究以大黄素为母体结构,人工半合成的4种蒽醌衍生物抑制KB和KBv200细胞增殖的作用机制.方法 采用MTT比色法测定细胞毒作用;采用流式细胞仪检测分别用DCFH-DA和DiOC6荧光探针标记的细胞内活性氧(ROS)和线粒体跨膜电位(△ψm).结果 4种蒽醌衍生物均能不同程度地抑制KB和KBv200细胞的增殖,表现为较强的体外抗肿瘤作用:1,8-二(二甲氨乙氨基)-3-甲基-6-甲氧基蒽醌(H-19)对KB和KBv200细胞IC50分别为1.37和1.42 μmol/L;1-吡啶乙氨基-3-甲基-6,8-二甲氧基蒽醌(H-21)的IC50分别是13.0和17.9 μmol/L;1-吡咯烷乙氨基-3-甲基-6,8-二甲氧基蒽醌(H-25)的IC50分别是8.5和11.7μmol/L;1-羟丁氨基-3-甲基-6,8-二甲氧基蒽醌(H-28)的IC50分别是7.6和8.6 μmol/L,且各药对敏感株KB和相应的多药耐药(MDR)细胞KBv200的IC50相近(P＞0.05).用4种药物分别处理两种细胞12、24、48 h,12 h即能引起ROS的明显增加,24 h达到最大,与48 h引起的ROS增加差异不显著;各药均引起两种细胞△ψm时间依赖性的降低,48h时△ψm降低达到最大.结论 4种大黄素蒽醌衍生物可能通过诱导细胞内ROS的增加,使得△ψm降低,从而抑制KB和KBv200细胞的生长,且对MDR细胞无抗药性,具有开发前景。

Object To study the inhibitory mechanism of four anthraquinone derivatives,which were obtained from modifying the structure of emodin,on proliferation of parental drug-sensitive KB cells and multidrug resistant ( MDR ) KBv200 cells.Methods Cytotoxicity was determined by tetrazolium (MTT)assay.Reactive oxygen species (ROS) levels and mitochondrialmembrane potential (△ψm ) in cells respectively labelled by DCFH-DA and DiOC6were assayed by flow cytometry.Results All four anthraquinone derivatives suppressed proliferation of KB and KBv200 cells,showed potent cytotoxicity,the mean IC₅₀ of H-19 (1,8-dimethylam inethylam ine-3-methyl-6-methoxy-anthraquinone) to KB and KBv200 cells was1.37 and 1.42μmol/L,respectively.IC₅₀ of H-21( 1-pyridylethylam ine-3-methyl-6,8-dimethoxy-anthraquinone ) was 13.0 and 17.9μmol/L,IC₅₀ of H-25 (1-pyrrolylethylam ine-3-methyl-6,8-dimethoxy-anthraquinone ) was 8.5 and 11.7μmol/L,IC₅₀ of H-28 (1-hydroxybutylam ine-3-methyl -6,8-dimethoxy-anthraquinone ) was 7.6 and 8.6μmol/L.The IC₅₀ of them to MDR KBv200 cellswas similar to that of them to the parental drug -sensitive KB cells ( P > 0.05).The generation of ROS increased obviously after the cells were incubated with them for 12 h,and the increase of ROS reached the peak treated for 24h.The levels of △ψm were timedependently decreased after treating with four compounds for 12,24,and 48 h.Conclusion The grouth of both MDR KBv200 cells and parental drug-sensitive KB cellswere inhibited to the treatment of four anthraquinone derivative in vitro.The mechanism of their effects is associated with the increase of the cellular ROS level and the decrease of △ψm.