苦参碱对K562及其多药耐药细胞K562/vin、K562/dox的诱导凋亡作用

Apoptosis of K562 Cell and Multidrug Resistant Cells(K562/vin,K562/dox) Induced by Matrine

目的 明确中药苦参碱对化疗敏感和耐药细胞的诱导凋亡作用。方法 采用MTT法测定苦参碱对各细胞的IC5 0值。K 5 6 2、K 5 6 2 /vin、K 5 6 2 /dox细胞与适宜浓度苦参碱共同孵育于 16 40培养液中 ,一定时间后对细胞行Wright′s Giemsa染色 ,做形态学观察 ,同时行DNA凝胶电泳、流式细胞仪DNA含量分析以检测凋亡。结果 细胞形态学观察可见 0 .2 5～ 1.0 0mg/ml浓度苦参碱作用后的细胞体积缩小、核固缩深染、细胞质空泡化、核碎裂等 ;DNA电泳可见阶梯状条带 ;流式细胞仪检测可见有低G1期细胞出现 ,并S期细胞比例增高。在试验范围内凋亡比率与药物浓度呈一定相关。苦参碱对敏感和耐药细胞的诱导凋亡作用无明显差异。结论 苦参碱可诱导K 5 6 2细胞凋亡 ,对其多药耐药细胞K 5 6 2 /vin、K 5 6 2 /dox亦有诱导凋亡作用 ,其作用与药物浓度呈一定相关性。苦参碱诱导肿瘤细胞凋亡可能与其S期阻滞有关。

Objective To investigate induction of apoptosis in matrine-treated chemotherapy sensitive cell line and multidrug-resistant(MDR) cell lines.Methods MTT assay was used to detect the half-inhibition concentration(IC50) of matrine on each cell line,then the matrine with appropriate concentration was chosen to treat K562、K562/vin、K562/dox cell for a certain time.Morphological changes of apoptotic cells were investigated by light microscopy.DNA fragmentation was analyzed by agarose gel electrophoresis.In addition,the number of apoptotic cells and the DNA content were quantitatively assessed by flow cytometry(FCM).Results 0.25～1.00 mg/ml matrine-treated K562、K562/Vin and K562/Dox cells all showed morphological characteristics of apoptosis such as contract of cell、condensation of nuclei、bubble of cytoplasm、fragment of nucleus.A ladder-like pattern of DNA fragmentation was demonstrated on agarose gel eletrophoresis.The DNA content analysis of FCM showed pre-G 1 cells and increased S period ratio.In the experiment range,the apoptosis-inducing effect of matrine was dose-dependent and showed no significant difference between sensitive cell and MDR cell.Conclusion Matrine can kill K562 cell by inducing the cells′ apoptosis and it also shows the effect of inducing apoptosis in MDR cell lines as K562/vin、K562/dox.The apoptosis effect is dose-dependent and it perhaps has certoin relation to the blocking of S period cells.