The effect of low doses of doxorubicin on the rat glioma C6 cells in the context of the proteins involved in intercellular interactions
The effect of low doses of doxorubicin on the rat glioma C6 cells in the context of the proteins involved in intercellular interactions
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DOI：
10.1016/j.acthis.2020.151625
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发表时间：
2020-10-01
影响因子：
2.5
通讯作者：
Grzanka, Alina
中科院分区：
文献类型：
--
作者：
Halas-Wisniewska, Marta;Izdebska, Magdalena;Grzanka, Alina
本研究的目的是确定多柔比星对大鼠胶质瘤C6细胞系中F-actin重排和β-catenin和cofilin-1的影响，并结合其形态学和超微结构的变化。实验材料为大鼠胶质瘤C6细胞系。将细胞与浓度为50、100和200 nM的亚致死剂量的多柔比星一起孵育。用蓝色台盼蓝染色法测定死亡细胞数。光镜和透射电镜观察细胞形态和超微结构的变化。为了确定F-肌动蛋白、β-连环蛋白和cofilin-1的重排和表达水平，使用荧光显微镜对其进行分析。进而，通过Guava 6 HT-2 L细胞仪评价细胞死亡和细胞周期。所进行的实验显示，用阿霉素处理后，C6细胞的存活率呈剂量依赖性降低。细胞死亡分析显示凋亡和坏死细胞数量呈剂量依赖性增加。这些结果通过显微镜观察得到证实。同时观察了细胞形态、超微结构的变化以及F-actin、beta-catenin和cofilin-1的重排。研究结果表明，亚致死浓度的阿霉素影响F-肌动蛋白和其他参与细胞间相互作用的蛋白质的结构。细胞凋亡可能先于有丝分裂的突变，提示低剂量阿霉素具有细胞毒作用。此外，我们的结果证实了DOX在肿瘤细胞中作用的多维机制。
The aim of this investigation was to determine the effect of doxorubicin on F-actin rearrangement and beta-catenin and cofilin-1 in a rat glioma C6 cell line in combination with changes in their morphology and ultrastructure. The experimental material constituted rat glioma C6 cell line. The cells were incubated with sublethal doses of doxorubicin in the concentration of 50, 100 and 200 nM. The blue trypan dye method was used to determine the number of dead cells. Morphological and ultrastructural changes in the cells were evaluated using light and transmission electron microscope, respectively. In order to determine the rearrangements and level of expression of F-actin, beta-catenin and cofilin-1 they were analyzed using a fluorecence microscope. In turn, cell death and cell cycle were evaluated by Guava 6HT-2 L Cytometer. The performed experiments showed a dose-dependent decrease in the survival of C6 cells after treatment with doxorubicin. The analysis of cell death showed a dose-dependent increase in the population of apoptotic and necrotic cells. These results were confirmed by microscopy observation. The changes in morphology, ultrastructure, and rearrangements of F-actin, beta-catenin and cofilin-1 were also observed. The results obtained in the study showed that sublethal concentrations of doxorubicin influenced the structure of F-actin and other proteins involved in cell-cell interactions. Moreover, mitotic catastrophe may preceding apoptosis, what suggest the cytotoxic effect of low dose of doxorubicin. Furthermore, our results confirmed the multi-dimensional mechanism of DOX action in tumor cells.