苦参碱对KB及其多药耐药细胞KBv200增殖与凋亡影响的对比研究

Comparative Study of the Effects of Matrine on the Proliferation and Apoptosis of Human Oral Epidermoid Carcinoma KB Cells and it's Moutidrug Resistant Counterpart KBv200 Cells

目的:研究中药苦参提取物苦参碱对体外培养的口腔上皮癌KB细胞和具有多药耐药特征的KBv200细胞增殖和凋亡的影响。方法:以MTT法对比观察苦参碱对细胞存活率的影响;以吖啶橙/溴乙啶(AO/EB)双荧光染色检测细胞凋亡率;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,其半数抑制浓度(IC50)分别为1.35mg/ml和1.43mg/ml;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。苦参碱对两种细胞增殖和凋亡的影响均未见明显差异(P>0.05)。结论:苦参碱对KB及多药耐药的KBv200细胞均具有抑制增殖和诱导凋亡的作用。提示苦参碱可克服肿瘤的多药耐药现象,故有可能成为一种理想的中药抗肿瘤制剂。

Objective: To evaluate the effects of Matrine on the proliferation and apoptosis of human oral epidermoid carcinoma KB cells and it's moutidrug resistant counterpart KBv200 Cells. Methods: The proliferation and apoptosis of KB Cells and KBv200 were studied comparatively after matrine treatment with the following methods. MTT assay was used to evaluate 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 as well as apoptosis of the two cells. Results: The vital rates of both KB and KBv200 cells in vitro were decreased after matrine treatment at different concentrations( 0.50、1.00、1.50、2.00 mg/ml). The IC50 concentrations of matrine on KB and KBv200 cells are 1.35 mg/ml and 1.43 mg/ml respectively, no sognificant difference was found between the two cells. The results of AO/EB fluorescent staining and flow cytometry showed that matrine could induce apoptosis of two cells. Under transmission electron microscope, condensation of nuclei, bubble of cytoplasm could be seen in both cell line in matrine treated cells. Matrine could block the growth of KB and KBv200 cells at S phase. Conclusion: Matrine could inhibit the proliferation and induce apoptosis of both KB and it's moutidrug resistant counterpart KBv200 cells. No differences were found in these effects between the two cells,suggesting that matrine could overcome multidrug resistance in the treatment of cancer and may of wider usage in oncological practice.