华西口腔医学杂志

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苦参碱对KB细胞及其多药耐药细胞KBv200的凋亡诱导作用

张金廷1,崔慧先2,李庆星1,张会军2,白 玉1   

  1. 1.河北医科大学第一医院 口腔科,河北 石家庄050031; 2.河北医科大学基础医学研究所 人体解剖学教研室,河北 石家庄050011
  • 收稿日期:2005-06-25 修回日期:2005-06-25 出版日期:2005-06-20 发布日期:2005-06-20
  • 通讯作者: 张会军, Tel: 0311-5917061
  • 作者简介:张金廷(1953-),男,河北人,教授,博士

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

ZHANGJin-ting1,CUIHui-xian2,LI Qing-xing1,ZHANGHui-jun1,BAI Yu2   

  1. 1.Dept.ofOral Medicine,The FirstHospital ofHebei Medical University,Shijiazhuang050031,China; 2.Dept.of Anatomy,Hebei Medical University,Shijiazhuang 050011,China
  • Received:2005-06-25 Revised:2005-06-25 Online:2005-06-20 Published:2005-06-20

摘要: 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、2·00 mg/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24 h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。

关键词: 苦参碱, 口腔上皮癌, KB细胞, 多药耐药, 凋亡

Abstract: Objective To investigate the induction of apoptosis on human oral epidermoid carcinoma KB cells and multidrug resistant KBv200 cells byMatrine.Methods MTT assay was used to investigate the inhibition ability of Matrine on the cellsin vitro. Transmission electronmicroscopewas used to observe the ultrastructure feature of cells aftertreated byMatrine. Acridine or- ange (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 cellswere decreased according toMatrine′s concentration. The IC50 concentrations ofMatrine on KB and KBv200 cells were 1.35 mg/ml and 1.43 mg/ml individually. The results of AO/EB fluores- cent staining and flow cytometry showed thatMatrine 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 byMatrine. 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.

Key words: matrine, human oral epidermoid carcinoma, KB cell, multidrug resistant, apoptosis