苦参碱对KB细胞及其多药耐药细胞KBv200的凋亡诱导作用

Apoptosis of Human Oral Epidermoid Carcinoma KB Cells and Multidrug Resistant KBv200 Cells Induced by Matrine

目的观察中药苦参的提取物苦参碱对口腔上皮癌KB细胞及其多药耐药细胞KBv200的凋亡诱导作用。方法MTT法观察苦参碱对体外培养的KB及KBv200细胞存活率的影响。吖啶橙溴乙啶(AO EB)双荧光染色检测二者细胞凋亡率。流式细胞术分析苦参碱对细胞周期的影响;扫描电镜观察苦参碱作用后的细胞形态学变化。结果苦参碱质量浓度在0.50、1.00、1.50、2.00mg ml时,均有抑制KB及KBv200增殖的作用,双荧光染色及流式细胞术提示苦参碱可诱导KB及KBv200细胞凋亡,使细胞生长停滞在S期;扫描电镜观察发现不同质量浓度的苦参碱作用24h后,细胞出现体积缩小、细胞质空泡化、细胞核碎裂等凋亡现象。结论苦参碱对KB及KBv200细胞有增殖抑制作用,可诱导KB及KBv200细胞凋亡,其机制可能与其导致细胞S期阻滞有关。

Objective To investigate the induction of apoptosis on human oral epidermoid carcinoma KB cells and multidrug resistant KBv200 cells by Matrine. Methods MTT assay was used to investigate the inhibition ability of Matrine on the cells in vitro. Transmission electron microscope was used to observe the ultrastructure feature of cells after treated by Matrine. Acridine orange (AO)/Ethidium bromide (EB) fluorescent staining and flow cytometry were used to observe apoptosis induced by Matrine. Flow cytometry was applied to study the effects of the drug on cell cycles of the cells. Results When 0.50,1.00,1.50,2.00 mg/ml of Matrine was used, the vital rates of KB and KBv200 cells were decreased according to Matrine′s concentration. The IC-{50} concentrations of Matrine on KB and KBv200 cells were 1.35 mg/ml and 1.43 mg/ml individually. The results of AO/EB fluorescent staining and flow cytometry showed that Matrine could induce apoptosis of two kinds of cells. While observed by transmission electron microscope, there were more contraction of cells, condensation of nuclei, bubble of cytoplasm in both kinds of cells after treated by Matrine. Matrine could stop the growth of KB and KBv200 cells at S period and restrain mitosis of cells. Conclusion Matrine can inhibit the growth of KB and KBv200 cells by inducing apoptosis. The apoptosis effect is dose_dependent and it has certain relation to the blocking of S period cells.