Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, Poland.
Acta Histochem. 2010;112(1):62-71. doi: 10.1016/j.acthis.2008.09.002. Epub 2008 Nov 11.
The purpose of this study was to assess whether Taxol-induced changes in microtubular dynamics are accompanied by apoptosis. CHO AA8 cells were treated with different Taxol concentrations (0.25microM, 0.5microM, 1microM) for 24h. The effects of Taxol exposure were analyzed using fluorescence microscopy and flow cytometry (TUNEL and annexin V-FITC/propidium iodide assays). 0.25microM Taxol caused the appearance of few multinucleated giant cells exhibiting extensive arrays of fine filaments. Slight increases in the level of polyploidy, phosphatidylserine externalization and in the percentage of TUNEL positive cells were noticed. Concentrations of 0.5 and 1microM resulted in the appearance of a large number of giant cells, which exhibited, depending on the cell, an extensive microtubular network or loose or tightly packed bundles of microtubules. Cells of reduced volume and showing chromatin condensation were also seen. Cell cycle analysis revealed that almost half of the cell population was polyploid. Except in cells exposed to 1microM Taxol, annexin V-FITC/PI labelling did not reveal the loss of plasma membrane integrity or increase in phosphatidylserine externalization; however, TUNEL assay revealed a significant increase in the percentage of cells with DNA fragmentation. These data indicate that CHO AA8 cells treated with Taxol undergo cell death of a type which considerably differs from apoptosis.
本研究旨在评估紫杉醇诱导的微管动力学变化是否伴随着细胞凋亡。CHO AA8 细胞用不同浓度的紫杉醇(0.25μM、0.5μM、1μM)处理 24 小时。用荧光显微镜和流式细胞术(TUNEL 和 Annexin V-FITC/碘化丙啶检测)分析紫杉醇暴露的影响。0.25μM 的紫杉醇导致少数多核巨细胞的出现,这些细胞表现出广泛的细丝阵列。多倍体水平、磷脂酰丝氨酸外翻和 TUNEL 阳性细胞比例略有增加。0.5μM 和 1μM 的浓度导致大量巨细胞的出现,这些细胞表现出广泛的微管网络或松散或紧密的微管束,这取决于细胞。还观察到体积减小和染色质浓缩的细胞。细胞周期分析显示,近一半的细胞群体为多倍体。除了暴露于 1μM 紫杉醇的细胞外,Annexin V-FITC/PI 标记未显示质膜完整性丧失或磷脂酰丝氨酸外翻增加;然而,TUNEL 检测显示 DNA 片段化的细胞比例显著增加。这些数据表明,用紫杉醇处理的 CHO AA8 细胞发生的细胞死亡类型与细胞凋亡有很大的不同。