Kurki P, Vanderlaan M, Dolbeare F, Gray J, Tan E M
Exp Cell Res. 1986 Sep;166(1):209-19. doi: 10.1016/0014-4827(86)90520-3.
The expression of proliferating cell nuclear antigen (PCNA), also called cyclin, was quantified in the cell lines SP2/0 and MOLT-4 and in mouse splenocytes induced to proliferate in vitro with mitogens. Autoantibody from a patient with systemic lupus erythematosus was used to label PCNA in cell suspensions after the cells had been fixed and permeabilized. In the same cells DNA was stained by propidium iodide. The cells were then analysed by flow cytometry for PCNA and DNA content. The PCNA profiles in proliferating spleen cells and the cell lines were similar. Most G0-G1 cells did not express significant amount of PCNA. A dramatic increase in PCNA immunofluorescence was observed in late G1 cells, and further increases were observed in S-phase cells. G2-M cells showed a reduced level of PCNA immunofluorescence relative to S-phase cells but were still elevated relative to G0-G1 cells. Proliferating cells arrested at the G1-S boundary by exposure to cytosine arabinoside showed an increased PCNA immunofluorescence as compared to unstimulated cells.
增殖细胞核抗原(PCNA),也称为细胞周期蛋白,在SP2/0和MOLT-4细胞系以及用有丝分裂原体外诱导增殖的小鼠脾细胞中进行了定量分析。在细胞固定并通透后,使用来自系统性红斑狼疮患者的自身抗体标记细胞悬液中的PCNA。在相同细胞中,用碘化丙啶对DNA进行染色。然后通过流式细胞术分析细胞的PCNA和DNA含量。增殖的脾细胞和细胞系中的PCNA图谱相似。大多数G0-G1期细胞不表达大量PCNA。在G1期晚期细胞中观察到PCNA免疫荧光显著增加,在S期细胞中进一步增加。相对于S期细胞,G2-M期细胞的PCNA免疫荧光水平降低,但相对于G0-G1期细胞仍有所升高。与未刺激的细胞相比,暴露于阿糖胞苷而停滞在G1-S边界的增殖细胞显示出PCNA免疫荧光增加。