Berntsson P S, Alm K, Oredsson S M
Department of Animal Physiology, Lund University, Helgonavägen 3 B, Lund, SE-223 62, Sweden.
Biochem Biophys Res Commun. 1999 Sep 16;263(1):13-6. doi: 10.1006/bbrc.1999.1293.
Cells in mitosis were seeded immediately after being harvested by the mitotic shake off technique from a culture of exponentially growing Chinese hamster ovary cells. At 2, 5, 7, 10, and 12 h after seeding, cycloheximide was added. Cells were sampled at various times after cycloheximide addition and the ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) activities were determined. Flow cytometric analysis showed that cells sampled at 2, 5, 7, 10, and 12 h after seeding were found in mid G(1), at the G(1)/S transition, in mid S phase, at the S/G(2) transition, and in late G(2), respectively. The half-lives of ODC and AdoMetDC activities varied during the cell cycle. The half-life of ODC activity showed a biphasic pattern with increases in connection to the G(1)/S and S/G(2) transitions while the half-life of AdoMetDC activity increased only at the G(1)/S transition.
采用有丝分裂振荡分离技术从指数生长期的中国仓鼠卵巢细胞培养物中收获处于有丝分裂期的细胞后,立即进行接种。接种后2、5、7、10和12小时,加入环己酰亚胺。在加入环己酰亚胺后的不同时间对细胞进行取样,并测定鸟氨酸脱羧酶(ODC)和S-腺苷甲硫氨酸脱羧酶(AdoMetDC)的活性。流式细胞术分析表明,接种后2、5、7、10和12小时取样的细胞分别处于G1中期、G1/S期转换点、S期中期、S/G2期转换点和G2后期。ODC和AdoMetDC活性的半衰期在细胞周期中有所不同。ODC活性的半衰期呈双相模式,在G1/S和S/G2期转换时增加,而AdoMetDC活性的半衰期仅在G1/S期转换时增加。