Mitotic cell death in BEL-7402 cells induced by enediyne antibiotic lidamycin is associated with centrosome overduplication
Mitotic cell death in BEL-7402 cells induced by enediyne antibiotic lidamycin is associated with centrosome overduplication
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DOI：
10.3748/wjg.v10.i18.2632
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发表时间：
2004-09-15
影响因子：
4.3
通讯作者：
Shao, Rong-Guang
中科院分区：
文献类型：
--
作者：
Liang, Yue-Xin;Zhang, Wei;Shao, Rong-Guang
目的：有丝分裂细胞死亡在肿瘤治疗中一直受到关注。然而，其背后的确切机制仍不清楚。我们以前曾报道过烯二炔类抗生素力达霉素在低浓度下诱导人上皮肿瘤细胞的有丝分裂细胞死亡。本研究旨在探讨中心体动力学与力达霉素诱导的人肝癌BEL-7402细胞有丝分裂死亡之间的可能联系。通过Hoechst 33342染色检测细胞多核化。流式细胞仪分析细胞周期。间接免疫荧光法检测异常中心体。蛋白质印迹和衰老相关的β-半乳糖苷酶（SA-β-gal）染色分别分析蛋白表达和衰老样表型。BEL-7402细胞暴露于低浓度的力达霉素导致含有多个中心体的细胞增加，这与有丝分裂细胞死亡的出现和某些细胞中SA-β-gal的活化有关，并伴有蛋白质表达的变化，调节细胞增殖和凋亡。线粒体信号通路是细胞凋亡的主要途径之一，在有丝分裂细胞死亡过程中没有被激活。异常的中心体有助于多极有丝分裂纺锤体的形成，这可能导致染色体的不平衡分裂和以多核或微核巨细胞为特征的有丝分裂细胞死亡。细胞周期分析显示，力达霉素可使BEL-7402细胞阻滞于G2/M期，这可能与中心体过度复制有关。结论：低浓度力达霉素可诱导BEL-7402细胞发生有丝分裂死亡和衰老。中心体调节异常可能在中等剂量力达霉素作用下有丝分裂失败和最终细胞死亡中起关键作用。
AIM: Mitotic cell death has been focused on in tumor therapy. However, the precise mechanisms underlying it remain unclear. We have reported previously that enediyne antibiotic lidamycin induces mitotic cell death at low concentrations in human epithelial tumor cells. The aim of this study was to investigate the possible link between centrosome dynamics and lidamycin-induced mitotic cell death in human hepatoma BEL-7402 cells.METHODS: Growth curve was established by MTT assay. Cell multinucleation was detected by staining with Hoechst 33342. Flow cytometry was used to analyze cell cycle. Aberrant centrosomes were detected by indirect immunofluorescence. Western blot and senescence-associated beta-galactosidase (SA-beta-gal) staining were used to analyze protein expression and senescence-like phenotype, respectively.RESULTS: Exposure of BEL-7402 cells to a low concentration of lidamycin resulted in an increase in cells containing multiple centrosomes in association with the appearance of mitotic cell death and activation of SA-beta-gal in some cells, accompanied by the changes of protein expression for the regulation of proliferation and apoptosis. The mitochondrial signaling pathway, one of the major apoptotic pathways, was not activated during mitotic cell death. The aberrant centrosomes contributed to the multipolar mitotic spindles formation, which might lead to an unbalanced division of chromosomes and mitotic cell death characterized by the manifestation of multi-or micronucleated giant cells. Cell cycle analysis revealed that the lidamycin treatment provoked the retardation at G2/M phase, which might be involved in the centrosome overduplication.CONCLUSION: Mitotic cell death and senescence can be induced by treatment of BEL-7402 cells with a low concentration of lidamycin. Centrosome dysregulation may play a critical role in mitotic failure and ultimate cell death following exposure to intermediate dose of lidamycin.