ORNITHINE DECARBOXYLASE AND S-ADENOSYLMETHIONINE DECARBOXYLASE EXPRESSION DURING THE CELL-CYCLE OF CHINESE-HAMSTER OVARY CELLS
ORNITHINE DECARBOXYLASE AND S-ADENOSYLMETHIONINE DECARBOXYLASE EXPRESSION DURING THE CELL-CYCLE OF CHINESE-HAMSTER OVARY CELLS
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DOI：
10.1006/excr.1995.1011
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发表时间：
1995-01-01
影响因子：
3.7
通讯作者：
OREDSSON, SM
中科院分区：
文献类型：
--
作者：
FREDLUND, JO;JOHANSSON, MC;OREDSSON, SM
采用有丝分裂分离技术，从指数生长的中国仓鼠卵巢细胞中收获处于有丝分裂期的细胞。收获后立即将有丝分裂细胞接种在组织培养瓶中，并在37 ℃下在CO2培养箱中孵育。注意不要干扰细胞在细胞周期中的进展。接种14 h后，每隔1 h收集细胞，分析细胞周期分布、细胞内多胺含量、鸟氨酸脱羧酶（ODC）和S-腺苷甲硫氨酸脱羧酶（S-adenosylmethionine decarboxylase，ODC）活性以及mRNA相对含量。通过DNA流式细胞术监测细胞周期的进展。腐胺、亚精胺和精胺水平在细胞周期中大约加倍：腐胺主要在S期和G期晚期（2），亚精胺在整个细胞周期中持续存在，精胺主要在G期（1）和S期。ODC活性在接种的有丝分裂细胞中较低，在G（1）晚期被激活，在S期达到平台。在S期晚期观察到活性的第二次爆发，并且在S/G（2）转换时发现最大ODC活性。ODC mRNA相对表达量在细胞周期中增加约一倍，主要发生在S期中后期。在G（1）晚期，MetDC活性增加，并且在G（1）/S转换期间观察到第一个最大值。在S期中期发现了第二次活动爆发。在G（2）中发现最大的α-MetDC活性。在细胞周期过程中，相对表达量增加约一倍，主要发生在G1期晚期和S期早期。我们的研究结果表明，多胺的合成在中国仓鼠卵巢细胞的细胞周期中的转录和翻译/翻译后水平的调节。(C)出版社：Academic Press
Cells in mitosis were harvested from exponentially growing Chinese hamster ovary cells by the mitotic detachment technique. Immediately after harvesting, the mitotic cells were seeded in tissue culture flasks and incubated at 37 degrees C in a CO2 incubator. Care was taken not to perturb the progression of cells through the cell cycle. At every hour after seeding for 14 h, cells were collected for analysis of cell cycle distribution, cellular polyamine content, ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) activities, and relative mRNA contents. The progression through the cell cycle was monitored by DNA flow cytometry. The putrescine, spermidine, and spermine levels were approximately doubled during the cell cycle: putrescine mainly during late S and G(2), spermidine continuously during the entire cell cycle, and spermine mainly during G(1) and S. The ODC activity was low in seeded mitotic cells and the enzyme was activated in late G(1) and reached a plateau in S phase. A second burst in activity was observed during late S phase and maximal ODC activity was found at the S/G(2) transition. The relative ODC mRNA level approximately doubled during the cell cycle and the increase in the relative level mainly took part during mid and late S phase. AdoMetDC activity increased in late G(1) and a first maximum was observed during the G(1)/S transition. A second burst in activity was found in mid S phase. Maximal AdoMetDC activity was found in G(2). The relative AdoMetDC mRNA approximately doubled during the cell cycle and the increase in the relative level mainly took place during late G(1) and early S phase. Our results indicate that polyamine synthesis was regulated at transcriptional and translational/post-translational levels during the cell cycle of Chinese hamster ovary cells. (C) 1995 Academic Press, Inc.