斑蝥素及去甲斑蝥素诱导人红白血病K562细胞凋亡的细胞学研究

STUDIES ON APOPTOTIC CYTOLOGY OF K562 HUMAN MYELOID LEUKEMIA CELLS INDUCED BY CANTHARIDIN AND NORCANTHARIDIN

目的 研究抗癌药物斑蝥素及去甲斑蝥素诱导人红白血病 K5 6 2细胞凋亡的细胞学变化特点。 方法 观察药物作用后 K5 6 2细胞形态的动态变化、一般及超微结构特征 ,并用流式细胞术分析斑蝥素及去甲斑蝥素诱导 K5 6 2细胞凋亡与细胞周期的关系。 结果 2 mg/ L斑蝥素及 10 mg/ L去甲斑蝥素作用 2 4h,部分 K5 6 2细胞质膜突起 ,染色质异常凝集 ,原位复染显示这些细胞质膜的通透性没有增强 ,DNA琼脂糖凝胶电泳呈现细胞凋亡特有的“梯子”状带纹。电镜下 ,加药处理组中部分细胞染色质凝集成块 ,没有核膜包被 ,部分细胞染色质靠核膜边集呈新月形 ,胞浆浓缩 ,内质网结构正常或有扩张 ,呈现典型的凋亡细胞形态。流式细胞术分析结合细胞形态学研究表明 ,斑蝥素及去甲斑蝥素诱导 K5 6 2细胞凋亡与细胞 G2 / M期阻滞相关。 结论 斑蝥素及去甲斑蝥素能够诱导 K5 6 2细胞凋亡 ,凋亡大部分发生在细胞周期的 M期 ,也有少量发生在间期 ,是多点启动的。

Objective\ Studying the apoptotic cytology of K562 cells induced by cantharidin(CA) and norcantharidin(NCTD).\ Methods\ Observing morphological changes and ultrastructural features of treated K562 cells by phase contrast microscope and electron microscope.Relation between apoptosis and cell cycle of treated K562 cells was examined by fluorescence\|activated cell sorting (FACS)analysis.\ Results\ Some K562 cells show cell membrane bubbling and nuclear chromatin compacting after treated with 2mg/L CA or 10mg/L NCTD for 24h.The membrane permeability of treated K562 cells proved no increase by counterstaining in situ.An apoptosis characteristic DNA“Ladder” detected by using agarose gel electrophoresis.Several masses of condensed chromatin without nuclear membrane were observed in some treated K562 cells,while chromatin of some other cells tended to marginate in crescents around the nuclear envelope with compacting cytoplasm,normal or slightly expanded endoplasmic reticulum,which presented typical morphology of apoptosis.FACS analysis of DNA profiles and cell morphological studies indicated that the apoptosis was related to tumor cell G\-2/M phase blocking.\ Conclusions\ Apoptosis was induced by CA and NCTD in K562 cells,most of which initiated in mitosis phase,others in interphase.