Roti Roti J L, Uygur N, Higashikubo R
Radiat Res. 1986 Aug;107(2):250-61.
Nuclear protein and DNA content of HeLa cells was determined as a function of time following hyperthermia by staining isolated nuclei with two fluorescent dyes: fluorescein isothiocyanate (FITC) for protein content and propidium iodide (PI) for DNA content. Bivariate FITC and PI histograms were obtained by flow cytometry. Univariate flow cytometric analysis was shown to be inadequate for this study, because some of the nuclear protein changes were due to cell cycle redistribution. Posthyperthermia cell kinetics could be divided into two distinct phases: an early phase characterized by the removal of heat-induced excess nuclear proteins with little or no cell progression through the cell cycle; and a late phase characterized by a redistribution of cells in the cell cycle resulting in an accumulation of cells in G2. The duration of these phases was dependent upon the hyperthermia dose. In the early phase, the rate of removal of excess nuclear protein was found to vary with heating time and temperature for time-temperature combinations which resulted in the same amount of excess nuclear protein. In the late phase, the cells blocked in G2 did not reduce their nuclear protein levels back to control values.
通过用两种荧光染料对分离的细胞核进行染色来测定热疗后不同时间点HeLa细胞的核蛋白和DNA含量：用异硫氰酸荧光素（FITC）测定蛋白含量，用碘化丙啶（PI）测定DNA含量。通过流式细胞术获得双变量FITC和PI直方图。单变量流式细胞术分析被证明不适用于本研究，因为一些核蛋白的变化是由于细胞周期重新分布所致。热疗后细胞动力学可分为两个不同阶段：早期阶段的特征是清除热诱导的过量核蛋白，细胞很少或没有通过细胞周期进展；晚期阶段的特征是细胞在细胞周期中重新分布，导致细胞在G2期积累。这些阶段的持续时间取决于热疗剂量。在早期阶段，发现对于导致相同量过量核蛋白的时间-温度组合，过量核蛋白的清除率随加热时间和温度而变化。在晚期阶段，阻滞在G2期的细胞没有将其核蛋白水平降低至对照值。
