Darzynkiewicz Z, Evenson D, Staiano-Coico L, Sharpless T, Melamed M R
Proc Natl Acad Sci U S A. 1979 Jan;76(1):358-62. doi: 10.1073/pnas.76.1.358.
A flow cytometric technique for simultaneous measurements of RNA and DNA in individual cells has been applied to correlate the content of cellular RNA with the rate of progression of cells through the S phase. Human peripheral lymphocytes stimulated with phytohemagglutinin were blocked at the G1/S phase boundary by hydroxyurea or 5-fluorodeoxy-uridine treatment. Cells in the G1 phase as well as cells blocked at the G1/S phase boundary showed high heterogeneity with respect to stainable RNA content. After release from the block, the cells traversed the S phase at rates proportional to the quantity of stainable RNA per cell. Cells with the highest RNA content completed DNA replication 5 hr after release from the block; the cells with minimal RNA traversed the S phase at one-fifth of this rate. The large intercellular variation in stainable RNA and length of the S phase may be due to functional heterogeneity in the lymphocyte population. Our results suggest a correlation between the number of ribosomes and the rate of DNA replication in lymphocytes.
一种用于同时测量单个细胞中RNA和DNA的流式细胞术已被应用于将细胞RNA含量与细胞通过S期的进程速率相关联。用植物血凝素刺激的人外周淋巴细胞通过羟基脲或5-氟脱氧尿苷处理被阻断在G1/S期边界。G1期的细胞以及被阻断在G1/S期边界的细胞在可染色RNA含量方面表现出高度异质性。从阻断中释放后,细胞以与每个细胞可染色RNA量成比例的速率穿过S期。RNA含量最高的细胞在从阻断中释放后5小时完成DNA复制;RNA最少的细胞以该速率的五分之一穿过S期。可染色RNA的细胞间差异大以及S期长度可能是由于淋巴细胞群体中的功能异质性。我们的结果表明淋巴细胞中核糖体数量与DNA复制速率之间存在相关性。