Darzynkiewicz Z, Evenson D P, Staiano-Coico L, Sharpless T K, Melamed M L
J Cell Physiol. 1979 Sep;100(3):425-38. doi: 10.1002/jcp.1041000306.
The metachromatic fluorochrome acridine orange was used to differentially stain DNA and RNA in Chinese hamster ovary (CHO) cells and in mitogen-stimulated human lymphocytes during their progression through the cell cycle. Green and red fluorescence of individual cells, representing cellular DNA and RNA, respectively, was measured by flow cytometry. CHO cells were synchronized by selective detachment at mitosis. Their rate of progression through G1 and subsequently through S phase correlated with the content of stainable RNA. The mean duration of the G1 phase was 5.2 hours for cells with high RNA content (highest 25 percentile population) and 8.1 hours for cells with low RNA (lowest 25 percentile). The duration of S phase was 5.9 and 7.5 hours for high- and low-RNA, 25 percentile subpopulations, respectively. Lymphocytes synchronized at the G1/S boundary by hydroxyurea or 5-fluorodeoxyuridine showed extremely high intercellular variation with respect to content of stainable RNA. After release from the block they traversed S phase at rates linearly proportional to the content of stainable RNA. The duration of S phase was five hours for cells with high RNA-, six to nine hours for cells with moderate RNA- and up to 27 hours for cells with minimal RNA-content. The data suggest that the rate of progression through the cell cycle of individual cells within a population may be correlated with the number of ribosomes per cell.
采用异染荧光染料吖啶橙对中国仓鼠卵巢(CHO)细胞以及有丝分裂原刺激的人淋巴细胞在细胞周期进程中进行DNA和RNA的鉴别染色。分别代表细胞DNA和RNA的单个细胞的绿色和红色荧光通过流式细胞术进行测量。CHO细胞通过在有丝分裂期选择性分离进行同步化。它们通过G1期以及随后通过S期的速率与可染色RNA的含量相关。对于RNA含量高的细胞(最高25%的群体),G1期的平均持续时间为5.2小时,而对于RNA含量低的细胞(最低25%),G1期的平均持续时间为8.1小时。对于RNA含量高和低的25%亚群,S期的持续时间分别为5.9小时和7.5小时。通过羟基脲或5-氟脱氧尿苷在G1/S边界同步化的淋巴细胞在可染色RNA含量方面表现出极高的细胞间差异。从阻断中释放后,它们以与可染色RNA含量成线性比例的速率穿过S期。对于RNA含量高的细胞,S期持续时间为5小时,对于RNA含量中等的细胞为6至9小时,对于RNA含量最低的细胞长达27小时。数据表明群体中单个细胞通过细胞周期的速率可能与每个细胞的核糖体数量相关。