Kuwahara M
Department of Neurosurgery, Nagasaki University.
Neurol Med Chir (Tokyo). 1991 Dec;31(13):853-8. doi: 10.2176/nmc.31.853.
The relationship between the thermosensitivity of cultured brain tumor cells and cytoskeleton was studied. C6 rat glioma cell line (C6 cells) and U-373-MG human glioblastoma cell line (MG cells) were used in monolayer culture. Survival rates at various temperatures were calculated by colony forming assay 10 days after heat treatment. Actin filaments, the main components of microfilaments, were observed by the 7-chloro-4-nitrobenzo-2-oxadiazole phallacidin staining and indirect immunofluorescence staining methods. Alpha-tubulins, the main components of microtubules, were also stained with an indirect immunofluorescence staining method. The morphological changes were investigated by scanning electron microscopy (SEM). Both the C6 cells and the MG cells showed moderate thermosensitivity on the survival curves. Actin filaments were revealed at stress fibers and the ruffles of the leading edge on both cell lines. Stress fibers were well developed in MG cells but were only minor in C6 cells. After heat treatment ruffles and stress fibers were disrupted. However, alpha-tubulins were not affected by heat treatment. SEM showed Swiss-cheese like change of cell surfaces due to many pores with disruption of ruffles and stress fibers after heat treatment. These results suggest that the cytoskeleton, especially microfilaments, may be damaged by hyperthermia.
研究了培养的脑肿瘤细胞热敏感性与细胞骨架之间的关系。采用单层培养的C6大鼠胶质瘤细胞系(C6细胞)和U-373-MG人胶质母细胞瘤细胞系(MG细胞)。热处理10天后,通过集落形成试验计算不同温度下的存活率。采用7-氯-4-硝基苯并-2-恶二唑鬼笔环肽染色和间接免疫荧光染色法观察微丝的主要成分肌动蛋白丝。微管的主要成分α-微管蛋白也采用间接免疫荧光染色法染色。通过扫描电子显微镜(SEM)研究形态学变化。C6细胞和MG细胞在存活曲线上均表现出中等热敏感性。在两种细胞系的应力纤维和前缘的褶皱处均发现了肌动蛋白丝。MG细胞中的应力纤维发育良好,而C6细胞中仅少量存在。热处理后,褶皱和应力纤维被破坏。然而,α-微管蛋白不受热处理的影响。SEM显示,热处理后,由于许多小孔以及褶皱和应力纤维的破坏,细胞表面呈现出类似瑞士奶酪的变化。这些结果表明,细胞骨架,尤其是微丝,可能会因热疗而受损。