CELL-CYCLE DEPENDENT EXPRESSION AND STABILITY OF THE NUCLEAR-PROTEIN DETECTED BY KI-67 ANTIBODY IN HL-60 CELLS
CELL-CYCLE DEPENDENT EXPRESSION AND STABILITY OF THE NUCLEAR-PROTEIN DETECTED BY KI-67 ANTIBODY IN HL-60 CELLS
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DOI：
10.1111/j.1365-2184.1992.tb01435.x
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发表时间：
1992-01-01
影响因子：
8.5
通讯作者：
DARZYNKIEWICZ, Z
中科院分区：
文献类型：
--
作者：
BRUNO, S;DARZYNKIEWICZ, Z
在人早幼粒细胞白血病HL-60细胞中研究了Ki-67抗体检测的增殖相关核抗原的表达和稳定性与其细胞周期进展的关系。这种抗原的表达是最小的G1晚期和早期S期细胞。抗原主要在S期在细胞中积累，并且在该期的后半期其每个细胞的增加速率加快。S期间Ki-67抗原的积累超过了DNA含量的增加，因此从G1晚期到G2 + M，Ki-67/DNA比率上升了80%。这种抗原从有丝分裂后的细胞中迅速消失。在长春碱诱导的有丝分裂后细胞中，估计该蛋白的半衰期似乎短于1小时。这种快速周转应与所研究细胞的G1期相对较长（6-8小时）的持续时间进行比较。在其中蛋白质从头合成被0.1 μ g/ml放线菌酮抑制的细胞中，也发现Ki-67抗原的半衰期为约1小时，与细胞周期中的细胞位置无关。因此，这些数据表明，在细胞周期中的这种蛋白质的水平的变化是其不同的合成速率，而不是其降解速率的阶段特异性变化的结果。由于晚期G1和极早期S期细胞仅以略高于背景的水平表达抗原，因此当使用Ki-67抗体作为细胞生长级分的标志物时，可能会将一些晚期G1细胞错误地分类为非循环细胞。
The expression and stability of the proliferation-associated nuclear antigen detected by Ki-67 antibody have been investigated in human promyelocytic leukaemic HL-60 cells in relation to their progression through the cell cycle. Expression of this antigen was minimal in late G1 and early S phase cells. The antigen accumulated in the cells predominantly during S phase, and its rate of increase per cell accelerated during the second half of this phase. The accumulation of Ki-67 antigen during S exceeded the increase in DNA content, and thus the Ki-67/DNA ratio rose 80% from late G1 to G2 + M. This antigen rapidly disappeared from post-mitotic cells. The half-life of this protein estimated in post-mitotic cells during stathmokinesis induced by vinblastine appeared to be shorter than 1 h. This rapid turnover should be compared with the relatively long (6-8 h) duration of G1 of the studied cells. In cells in which de novo protein synthesis was inhibited by 0.1-mu-g/ml cycloheximide, the half-life of the Ki-67 antigen was also found to be about 1 h regardless of the cell position in the cell cycle. Thus, the data suggest that variations in the level of this protein during the cell cycle are a consequence of its different synthesis rate rather than phase-specific changes in the rate of its degradation. Because the late G1 and very early S phase cells express the antigen at levels only slightly above background, it is possible that, when using Ki-67 antibody as a marker of the cell growth fraction, some late G1 cells can be erroneously classified as non-cycling cells.