Roti Roti J L, Higashikubo R, Blair O C, Uygur N
Cytometry. 1982 Sep;3(2):91-6. doi: 10.1002/cyto.990030205.
To determine the change in nuclear protein content as a function of cell cycle position, isolated HeLa nuclei were stained for protein with fluorescein isothiocyanate (FITC) and for DNA with propidium iodide (PI) and analyzed by flow cytometry (FCM). The resulting FITC versus PI histogram consisted of four definable regions, a G1 region characterized by increasing FITC and relatively constant PI (2C DNA content), an S region characterized by increasing PI with relatively constant FITC, a G2 region characterized by increasing FITC and constant PI (4C DNA content), and a region of G1 FITC staining with near G2 PI staining. The relationship between cell cycle position and these regions of the histogram was confirmed by the two following studies: 1) The distribution of labeled nuclei throughout the histogram was observed after [14C]TdR pulse labeling. 2) Exit of cells from G1 was observed in the histogram after the addition of Colcemid to the HeLa cell cultures. Nuclear protein content did not appear to increase uniformly across the cell cycle (defined by DNA content). Rather, nuclear protein content showed the largest increase during G1. Thus, dual parameter FCM analysis based on nuclear DNA and protein content provides a more complete definition of cell cycle position than DNA content alone.
为了确定核蛋白含量随细胞周期位置的变化，将分离的HeLa细胞核用异硫氰酸荧光素（FITC）进行蛋白质染色，用碘化丙啶（PI）进行DNA染色，并通过流式细胞术（FCM）进行分析。所得的FITC与PI直方图由四个可定义区域组成，一个G1区域，其特征是FITC增加而PI相对恒定（2C DNA含量）；一个S区域，其特征是PI增加而FITC相对恒定；一个G2区域，其特征是FITC增加而PI恒定（4C DNA含量）；以及一个具有接近G2 PI染色的G1 FITC染色区域。细胞周期位置与直方图这些区域之间的关系通过以下两项研究得到证实：1）在[14C]TdR脉冲标记后观察标记细胞核在整个直方图中的分布。2）在向HeLa细胞培养物中添加秋水仙酰胺后，在直方图中观察到细胞从G1期退出。核蛋白含量在整个细胞周期（由DNA含量定义）中似乎并非均匀增加。相反，核蛋白含量在G1期增加最大。因此，基于核DNA和蛋白含量的双参数FCM分析比单独的DNA含量能更完整地定义细胞周期位置。
