Hang H, Fox M H
Department of Radiological Health Sciences, Colorado State University, Fort Collins 80523, USA.
Cytometry. 1996 Dec 1;25(4):367-73. doi: 10.1002/(SICI)1097-0320(19961201)25:4<367::AID-CYTO8>3.0.CO;2-A.
Seven different cell lines were analyzed by flow cytometry to evaluate the variation in heat shock protein hsp70 through the cell cycle. Inducible (hsp72) or both constitutive and inducible (hsp70) heat shock proteins were measured with monoclonal antibodies, and the cell cycle distribution was simultaneously measured with propidium iodide. Cell lines analyzed were Rat-1, Hr-24 (Rat-1 transfected with human hsp 72 gene), CHO (Chinese hamster ovary), C3H-10T1/2, AG1522 (normal human foreskin), GM2149 (normal human female skin), and Hela. None of the cell lines had a substantial variation in hsp72 or hsp70 levels through the cell cycle if they were not heated. In contrast, after chronic heating at 42.0 degrees C for 7.5 h, different cell lines had different patterns of hsp72 or hsp70 through the cell cycle. These results demonstrate that the level of hsp70 is not regulated differentially through the cell cycle in a variety of mammalian cell lines under normal unheated conditions. However, heat shock does induce cell-cycle-specific regulation of hsp70, which varies for different cell lines.
通过流式细胞术分析了七种不同的细胞系,以评估热休克蛋白hsp70在细胞周期中的变化。用单克隆抗体测量诱导型(hsp72)或组成型和诱导型(hsp70)热休克蛋白,并同时用碘化丙啶测量细胞周期分布。所分析的细胞系包括Rat-1、Hr-24(转染了人hsp 72基因的Rat-1)、CHO(中国仓鼠卵巢细胞)、C3H-10T1/2、AG1522(正常人包皮细胞)、GM2149(正常人类女性皮肤细胞)和Hela细胞。如果不加热,没有一个细胞系的hsp72或hsp70水平在细胞周期中存在显著变化。相反,在42.0℃慢性加热7.5小时后,不同细胞系在细胞周期中hsp72或hsp70呈现不同模式。这些结果表明,在正常未加热条件下,多种哺乳动物细胞系中hsp70的水平在细胞周期中没有差异调节。然而,热休克确实会诱导hsp70的细胞周期特异性调节,不同细胞系的调节方式有所不同。