Evidence for cell cycle phase-specific initiation of a program of HL-60 cell myeloid differentiation mediated by inducer uptake.
Evidence for cell cycle phase-specific initiation of a program of HL-60 cell myeloid differentiation mediated by inducer uptake.
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发表时间：
1984-06
期刊：
影响因子：
11.2
通讯作者：
A. Yen;K. Albright
中科院分区：
文献类型：
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作者：
A. Yen;K. Albright
利用人早幼粒白血病细胞株HL-60研究了最初的调控事件是否具有细胞周期特异性的问题，该调控事件引导未决定的前体细胞走向终末分化。虽然HL-60系统没有反映正常造血的所有特征，但它确实提供了一个相对定义明确的体外实验系统，可以用于检查分化过程的各个方面。用维甲酸诱导HL-60细胞向髓系分化。测定HL-60细胞在不同细胞周期阶段的分化动力学。这与细胞摄取维甲酸作为细胞周期位置的函数进行了比较。如果初始分化调节事件是细胞周期阶段独立的，那么分化动力学将独立于初始种群的细胞周期状态。基于细胞窄角和正交光散射强度谱的流式细胞仪细胞分选方法，在没有药物干扰的情况下，筛选出G1富集型和S+G2+M富集型细胞群。用10(-6)Mβ-全反式维甲酸诱导这两个群体进行髓系分化。在这两个群体中，都测量了与末端细胞分化相关的G1/0停滞的动力学，以及通过氧化代谢发展来分析表型分化的动力学。两个种群的分化动力学不同，表明最初的分化调节事件是细胞周期特异性的。对于两个初始细胞群体，在S期细胞的相对数量增加约24小时后，显著的表型分化。当指数增殖期的HL-60细胞暴露于10(-5)M[~(-5)M]维甲酸1小时后，流式细胞术分析发现，S+G2+M期细胞的摄取是G1期和S期细胞的近10倍。结果表明，当摄取增强时，细胞髓系分化的调控首先对诱导剂维甲酸起反应。
The question of whether the initial regulatory event, which directs an uncommitted precursor cell toward terminal differentiation, is cell cycle phase specific was examined using the human promyelocytic leukemia cell line, HL-60. While the HL-60 system does not reflect all of the features of normal hematopoiesis, it does provide a relatively well-defined in vitro experimental system which can be useful for examining aspects of the differentiation process. HL-60 cells were induced to undergo myeloid differentiation by retinoic acid. The subsequent differentiation kinetics of HL-60 populations initially enriched in different cell cycle phases was measured. This was compared to the cellular uptake of retinoic acid as a function of cell cycle position. If the initial differentiation-regulating event were cell cycle phase independent, then the kinetics of differentiation would be independent of the cell cycle status of the initial population. Flow cytometric cell sorting, based on cellular narrow angle and orthogonal light scatter intensity spectra, was used to select G1-enriched and S + G2 + M-enriched cell populations without pharmacological perturbation. These two populations were each induced to undergo myeloid differentiation with 10(-6) M beta-all-trans-retinoic acid. The kinetics of G1/0 arrest associated with terminal cell differentiation, as well as phenotypic differentiation, assayed by development of oxidative metabolism, was measured for both populations. The kinetics of differentiation differed for the two populations, indicating that the initial differentiation-regulating event was cell cycle phase specific. For both of the initial cell populations, significant phenotypic differentiation followed approximately 24 hr after enrichment in the relative number of S-phase cells. When exponentially proliferating HL-60 cells were exposed to a 1-hr pulse of 10(-5) M [3H]retinoic acid and then flow cytometrically sorted by DNA content, cells in late S + G2 + M had an approximately 10-fold higher uptake than cells in G1 or early S. The results indicate that cellular regulation of myeloid differentiation first becomes responsive to the inducer, retinoic acid, in S phase when uptake is enhanced.