Li G C, Petersen N S, Mitchell H K
Int J Radiat Oncol Biol Phys. 1982 Jan;8(1):63-7. doi: 10.1016/0360-3016(82)90386-8.
We have performed experiments to determine the kinetics of induction of thermal tolerance in Chinese hamster HA-1 cells, and the effects of heat treatments on the recovery of protein synthesis, with particular attention to whether heat induces specific proteins, perhaps the heat shock proteins (HSP). The kinetics of the development of thermal tolerance were measured by increases in cellular survival. In parallel experiments, the effects of heat treatment on the recovery of protein synthesis in HA-1 cells were examined. After heating (45 degrees, 20 minutes), some of these cells were immediately labeled with 35S-methionine (10 microCi/ml) for 1 hour at 37 degrees, while the others were incubated at 37 degrees for 1-8 hours and then labeled. The cell samples were prepared for electrophoresis on a gradient SDS gel. The incorporation of label into HA-1 cell proteins was drastically inhibited by the 45 degrees heat treatment, but recovered gradually during the 8-hour incubation period at 37 degrees C. A comparison of the proteins synthesized following heat shock with those synthesized by non-heated cells showed that the levels of synthesis of certain proteins were greatly enhanced following the 45 degrees treatment. By 8 hours, it was qualitatively apparent that three proteins, with molecular weights of 59K, 70K and 87K, were synthesized in greater amounts than in untreated cells. The kinetics of HSP synthesis were compared to the kinetics of thermal tolerance; these showed good correlation. Overall protein synthesis also increased during this time, although at a rate slower than the synthesis of the HSP. The question of whether the HSP play a causative role in the development of thermal tolerance and if so, what role might be, has not been answered.
我们进行了实验，以确定中国仓鼠HA-1细胞中热耐受性诱导的动力学，以及热处理对蛋白质合成恢复的影响，特别关注热是否诱导特定蛋白质，可能是热休克蛋白（HSP）。通过细胞存活率的增加来测量热耐受性发展的动力学。在平行实验中，研究了热处理对HA-1细胞中蛋白质合成恢复的影响。加热（45摄氏度，20分钟）后，一些细胞立即用35S-甲硫氨酸（10微居里/毫升）在37摄氏度下标记1小时，而其他细胞在37摄氏度下孵育1-8小时后再进行标记。细胞样品制备好后在梯度SDS凝胶上进行电泳。45摄氏度的热处理极大地抑制了HA-1细胞蛋白质中标记的掺入，但在37摄氏度的8小时孵育期内逐渐恢复。将热休克后合成的蛋白质与未加热细胞合成的蛋白质进行比较，结果表明，45摄氏度处理后某些蛋白质的合成水平大大提高。到8小时时，定性地明显看出，三种分子量分别为59K、70K和87K的蛋白质合成量比未处理细胞中的更多。将HSP合成的动力学与热耐受性的动力学进行了比较；结果显示两者具有良好的相关性。在此期间，总体蛋白质合成也有所增加，尽管其速率比HSP的合成速率慢。HSP在热耐受性发展中是否起因果作用，如果是，可能起什么作用，这个问题尚未得到解答。
