Department of Histology and Embryology, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University, Poland.
Acta Histochem. 2013 Jan;115(1):8-15. doi: 10.1016/j.acthis.2012.02.006. Epub 2012 Apr 6.
Hyperthermia is used in cancer therapy, however much remains to be discovered regarding its mechanisms of action at the cellular level. In this study, the effects of hyperthermia on cell death, survival, morphology and the cytoskeleton were investigated in a non-small cell lung cancer cell line, H1299. Despite the fact that this cell line is widely used in research, it has not yet been tested for heat shock sensitivity. Cells were given a 30-min heat shock at 43.5°C and 45°C and left to recover at 37°C for 24 and 48 h. 24 h after heat shock treatment, we monitored changes in the organization of the cytoskeleton using immunofluorescence microscopy. The number of actin stress fibers was significantly reduced, microtubules formed a looser meshwork, a portion of the cells possessed multipolar mitotic spindles, whereas vimentin filaments collapsed into perinuclear complexes. 48 h following heat stress, most of the cells showed recovery of the cytoskeleton, however we observed a considerable number of giant cells that were multinucleated or contained one enlarged nucleus. The data obtained by MTT assay showed a dose-dependent decrease of cell viability, while flow cytometric analysis revealed an increase in the number of cells with externalized phosphatidylserine. The results suggest that one of the modes of heat-induced cell death in H1299 cells is mitotic catastrophe, which probably ends in apoptosis.
热疗被用于癌症治疗，但在细胞水平上，其作用机制仍有许多有待发现。在这项研究中，我们研究了非小细胞肺癌细胞系 H1299 中热疗对细胞死亡、存活、形态和细胞骨架的影响。尽管该细胞系广泛应用于研究，但尚未对其热休克敏感性进行测试。细胞在 43.5°C 和 45°C 下接受 30 分钟的热冲击，然后在 37°C 下恢复 24 和 48 小时。在热休克处理 24 小时后，我们使用免疫荧光显微镜监测细胞骨架的组织变化。肌动蛋白应力纤维的数量明显减少，微管形成更松散的网格，一部分细胞具有多极有丝分裂纺锤体，而波形蛋白丝塌陷成核周复合物。热应激后 48 小时，大多数细胞的细胞骨架恢复，但我们观察到相当数量的巨细胞具有多核或一个增大的核。MTT 测定得到的结果表明细胞活力呈剂量依赖性下降，而流式细胞术分析显示，具有外翻磷脂酰丝氨酸的细胞数量增加。结果表明，H1299 细胞中热诱导细胞死亡的模式之一是有丝分裂灾难，这可能以细胞凋亡告终。
