Pollack A, Bagwell C B, Irvin G L, Jensen J A
Cytometry. 1980 Jul;1(1):57-64. doi: 10.1002/cyto.990010112.
Flow cytometry (FCM) was used to monitor the radiation effects promoted by incorporated tritiated thymidine (3H-TdR) on phytohemagglutinin (PHA)-stimulated human peripheral blood lymphocytes stained with propidium iodide (PI). Lymphocyte microcultures were continuously labeled or pulse-labeled for various periods of time with different 3H-TdR concentrations. Two types of DNA histogram analyses were performed on unperturbed and 3H]TdR perturbed lymphocytes. The data analyses consisted of statistical analyses between averaged groups of histograms (nonparametric analysis) and cell cycle analyses (parametric analysis) to determine the percentages of cells in G0 + G1, S and G2 + M. The results showed that (a) 3H-TdR when added to proliferating lymphocytes under certain conditions (both short-term continuous and pulse-labeling) caused a highly significant increase in the proportion of tetraploid (4C) cells by FCM, (b) the increase in the proportion of 4C cells represented a block in G2 and (c) the relative increase in the percentage of 4C cells was proportional to 3H-TdR incorporation which was proportional to labeling time and concentration. Therefore, it was concluded that short labeling times be used to minimize adverse radiation effects when 3H-TdR is used to assay substances affecting lymphocyte proliferation or in the estimation of cell cycle time.
采用流式细胞术（FCM）监测掺入的氚标记胸腺嘧啶核苷（3H-TdR）对用碘化丙啶（PI）染色的植物血凝素（PHA）刺激的人外周血淋巴细胞的辐射效应。淋巴细胞微培养物用不同浓度的3H-TdR连续标记或脉冲标记不同时间。对未受干扰和受3H-TdR干扰的淋巴细胞进行了两种类型的DNA直方图分析。数据分析包括直方图平均组之间的统计分析（非参数分析）和细胞周期分析（参数分析），以确定处于G0+G1、S和G2+M期的细胞百分比。结果表明：（a）在某些条件下（短期连续标记和脉冲标记），当将3H-TdR添加到增殖的淋巴细胞中时，通过FCM检测到四倍体（4C）细胞比例显著增加；（b）4C细胞比例的增加代表G2期阻滞；（c）4C细胞百分比的相对增加与3H-TdR掺入量成正比，而3H-TdR掺入量与标记时间和浓度成正比。因此得出结论，当使用3H-TdR检测影响淋巴细胞增殖的物质或估计细胞周期时间时，应采用短标记时间以尽量减少不良辐射效应。
