三氮唑席夫碱衍生物诱导SMMC-7721细胞分化为多倍体巨细胞  

The induction of SMMC-7721 cells into polyploid giant cells using triazole schiff base derivative in vitro

作　　者：皇甫超申[1] 刘彬[1] 马永超[1] 胡国强[1] 

机构地区：[1]河南大学神经生物学研究所河南大学医学院

出　　处：《解剖学报》2009年第1期92-97,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30670688,30771140)

摘　　要：目的探讨三氮唑席夫碱衍生物（LH-37）诱导SMMC．7721细胞形成多倍体巨细胞的作用。方法SMMC-7721细胞以1×10^4／ml的浓度接种于含1×10^-5mol／LLH．37的培养液内72h，流式细胞术分析DNA含量，Hoechst3258和碘化丙啶双染观察细胞死亡，免疫荧光检查微管和核纤层构造，透射电镜观察巨细胞超微结构。结果LH-37作用SMMC-7721细胞72h，巨细胞明显增加，单核巨细胞和多核巨细胞分别为12．32％和0．92％。可见多核巨细胞子核放射状排列，微管增厚并包裹子核，形成单-的微管组织中心，放射状排列的子核被完整的核纤层分割包绕，大部分多核巨细胞凋亡。单核巨细胞的微管常围绕核膜凝集，同时向细胞膜方向放射状分布，可见异常的有丝分裂和凝集的染色质，电镜下可见-些单核巨细胞凋亡时伴有自吞噬特征。结论LH-37通过干扰微管动力学，导致多倍体巨细胞增加和延迟性细胞死亡。Objective To analyze the features of polyploid giant cells that come from SMMC-7721 cells after being induced by triazole Schiff base derivative （LH-37） in vitro. Methods The SMMC-7721 cells （1×10^4/ml） were cultivated in medium containing of 1 ×10^-5 mol/L LH-37 for 72 hours, and their DNA content was analyzed by flow eytometry. The Co-staining with Hoechst 33258 and propidium iodide were used to observe cell death. Behavior of the microtubule（MTs） and lamin-B configuration apparatus in cells were examined by immunofluorescence, counterstained with Hoest33258 staining,and the uhrastrueture of giant tumor cells was observed by JEM 100CX-Ⅱ electron microscopes. Results Increased the proportion of mono-nucleated or multi- nucleated giant cells （MOGC and MNGC） after SMMC-7721 cells were cultured in the presence of LH-37 at 72 hours. The proportion of the giant cells in MOGC and MNGC was 12.32% and 0.92% respectively. Arrangement and radial segregation of sub-nuclei, sub-nuclei were united through MTs which radically emerged and mnbraced them from a single microtubule organizing centre （MTOC）, thick microtubule tracks reached individual nucleoli; sub-nuclei of this radial arrangement was encircled by the intact lamin; and most of the MNGCs died by apoptotic crisis. MTs of MOGC uniformly spread throughout the cytoplasm from the nuclear envelope and usually accumulated in the perinuelear region. Abnormal mutipolar mitosis, and spontaneous chromosome condensation were observed. In addition electron micrographs demonstrated features of autophagy during apoptosis of some MOGCs. Conclusion The data presented indicate that LH-37 （through the disturbance of microtubule dynamics）, can lead to an increase in polyploid giant cells and causative delayed death.

关 键 词：三氮唑席夫碱衍生物 肝癌细胞 多倍体巨细胞 微管 免疫荧光 人 

分 类 号：R735.2[医药卫生—肿瘤]