Ninomiya-Tsuji J, Nakahara Y, Ito C, Akiyama T, Ishibashi S, Ide T
Department of Physiological Chemistry, Hiroshima University, School of Medicine, Japan.
Cell Struct Funct. 1987 Oct;12(5):421-32. doi: 10.1247/csf.12.421.
tsJT60, a temperature-sensitive (ts) G0-mutant cell line from a Fischer rat, grows normally in the exponential growth phase at 34 degrees C and 39.5 degrees C, but when stimulated with fetal bovine serum (FBS), from the G0 phase they reenter the S phase at 34 degrees C but not at 39.5 degrees C. The ts-block was bypassed when G0-arrested tsJT60 cells were stimulated at 39.5 degrees C with FBS plus epidermal growth factor (EGF). The presence of EGF for the first 6 h after serum stimulation caused tsJT60 cells to enter the S phase in the presence of FBS at 39.5 degrees C. When EGF was added 6 h after serum stimulation, entrance into the S phase was delayed by about 6 h. The sequential presence of two growth factors, EGF without FBS for 6 h then FBS without EGF, or the reversed sequence, failed to initiate DNA synthesis at 39.5 degrees C. The binding of EGF was not temperature sensitive. The amounts of RNA and protein present doubled after stimulation with both FBS and EGF at 39.5 degrees C. These and other findings suggest that EGF bypasses only some specific event in the entire prereplicative process that operates operating in serum-stimulated cells at 39.5 degrees C.
tsJT60是一种来自Fischer大鼠的温度敏感(ts)G0突变细胞系,在34摄氏度和39.5摄氏度的指数生长期能正常生长,但在用胎牛血清(FBS)刺激时,它们在34摄氏度时从G0期重新进入S期,而在39.5摄氏度时则不会。当用FBS加表皮生长因子(EGF)在39.5摄氏度刺激G0期停滞的tsJT60细胞时,ts阻断被绕过。血清刺激后最初6小时内存在EGF,会使tsJT60细胞在FBS存在的情况下于39.5摄氏度进入S期。当在血清刺激6小时后添加EGF时,进入S期会延迟约6小时。两种生长因子按顺序存在,即先在无FBS的情况下添加EGF 6小时,然后在无EGF的情况下添加FBS,或者顺序相反,都无法在39.5摄氏度启动DNA合成。EGF的结合不具有温度敏感性。在39.5摄氏度用FBS和EGF刺激后,RNA和蛋白质的含量翻倍。这些以及其他发现表明,EGF仅绕过了在39.5摄氏度受血清刺激的细胞中整个复制前过程中的某些特定事件。