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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1×104/ml的浓度接种于含1×10-5mol/L LH-37的培养液内72h,流式细胞术分析DNA含量,Hoechst33258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,形成单一的微管组织中心,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,电镜下可见一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 investigate the effect of the Schiff base derivative of triazole (LH-37) on the formation of polyploid giant cells in SMMC-7721 cells. Methods SMMC-7721 cells were inoculated into culture medium containing 1 × 10-5 mol / L LH-37 at a concentration of 1 × 104 / ml for 72 hours. The DNA content was analyzed by flow cytometry. Hoechst33258 and propidium iodide double staining cells Death, immunofluorescence microtubules and lamina structure, transmission electron microscopy of the ultrastructure of giant cells. Results LH-37 treated SMMC-7721 cells 72h, the giant cells increased significantly, mononuclear giant cells and multinucleated giant cells were 12.32% and 0.92%. It can be seen that the multinucleated giant cell nuclei are radially arranged, the microtubules are thickened and enveloping the nucleus to form a single microtubule tissue center. The radially arranged nuclei are surrounded by a complete nuclear lamina and most of the multinucleated giant cells are apoptotic. Mononuclear giant cell microtubules often around the nuclear membrane agglutination, while radial distribution to the cell membrane, showing abnormal mitosis and aggregation of chromatin, electron microscopy can be found in some mononuclear giant cell apoptosis with autophagy characteristics. Conclusions LH-37, by interfering with microtubule dynamics, leads to polyploid giant cell hyperplasia and delayed cell death.