Fuse T
Department of Neurosurgery, Nagoya City University Medical School, Japan.
No To Shinkei. 1991 Sep;43(9):843-50.
Expression of major heat shock and stress-induced protein, HSP70, is known to be under complex regulation in tumor cells. In this study, we investigated the alternations of cytokinetics and HSP70 expression by hyperthermia in the in vitro experimental systems, using two rat glioma cell lines, two human glioblastoma cell lines and rat glioblast cells. For hyperthermal treatment the flasks were placed in water baths warmed up at 41 -45 degrees C for 15 min. To determine the effect of hyperthermia on the cell cycle progression, the changes in the DNA distribution of the cell population were studied by flow cytometry (FCM). The levels of HSP70 protein were determined by immunoblot analysis. The relationship between cell cycle and HSP70 expression was investigated by FCM using PI and FITC-labelled HSP70 double staining technique. These results were as follows: 1) Compared with the control, hyperthermic treatment at 42 degrees C or 44 degrees C caused both 354A and T98G cells to accumulate in S phase 18 hours after treatment and G2/M phase after 6-18 hours. 2) Hyperthermic treatment at 42 degrees C caused C6 cells to accumulate in S phase 6 hours after treatment, whereas heat treatment at 44 degrees C caused C6 cells to accumulate in S phase after 18 hours and G2/M phase after 6 hours. 3) A172 cells were accumulated only in G2/M phase by hyperthermia. 4) Glioblast cells did not show the alterations of cytokinetics by heat treatment remarkably. 5) HSP70 protein synthesis were enhanced under hyperthermic conditions in all type of cells, whether primary glioblast or permanent glioma cell lines.(ABSTRACT TRUNCATED AT 250 WORDS)
已知主要热休克蛋白和应激诱导蛋白HSP70在肿瘤细胞中的表达受到复杂调控。在本研究中,我们使用两种大鼠胶质瘤细胞系、两种人胶质母细胞瘤细胞系和大鼠胶质母细胞,在体外实验系统中研究了热疗对细胞动力学和HSP70表达的影响。对于热疗,将培养瓶置于41 - 45摄氏度的水浴中加热15分钟。为了确定热疗对细胞周期进程的影响,通过流式细胞术(FCM)研究细胞群体DNA分布的变化。通过免疫印迹分析确定HSP70蛋白水平。使用PI和FITC标记的HSP70双重染色技术,通过FCM研究细胞周期与HSP70表达之间的关系。结果如下:1)与对照组相比,42摄氏度或44摄氏度的热疗使354A和T98G细胞在处理后18小时积聚在S期,6 - 18小时后积聚在G2/M期。2)42摄氏度的热疗使C6细胞在处理后6小时积聚在S期,而44摄氏度的热处理使C6细胞在18小时后积聚在S期,6小时后积聚在G2/M期。3)热疗使A172细胞仅积聚在G2/M期。4)胶质母细胞经热处理后细胞动力学未出现明显改变。5)在热疗条件下,所有类型的细胞,无论是原代胶质母细胞还是永久性胶质瘤细胞系,HSP70蛋白合成均增强。(摘要截短至250字)