Barker K A, Orkin S H, Newburger P E
Department of Pediatrics, University of Massachusetts Medical School, Worcester 01655.
Mol Cell Biol. 1988 Jul;8(7):2804-10. doi: 10.1128/mcb.8.7.2804-2810.1988.
The expression of the X-CGD gene, which encodes the heavy-chain subunit of the phagocyte cytochrome b, was studied during induced myeloid differentiation of HL-60 cells. Incubation of the cells with a combined regimen of retinoic acid and dimethyl formamide resulted in granulocytic morphological differentiation and acquisition of nitroblue tetrazolium reduction, a measure of superoxide generation. During the 5-day course of induced differentiation, the levels of X-CGD mRNA transcripts rose 13-fold, with a 2-fold increase detectable within 3 h of exposure to retinoic acid. Relative transcription rates for the X-CGD gene, determined by nuclear runoff, increased two- to eightfold after 24 to 72 h of induced differentiation. However, the greater change in X-CGD mRNA levels than that in transcription rates implies the involvement of posttranscriptional regulation as well. Fractionation by centrifugal elutriation into phases of the cell cycle showed expression of X-CGD transcripts predominantly in G1 cells before induction and in all phases of the cell cycle 24 h after induction. Thus the rapid increase in X-CGD expression in induced cells reflects the acquisition of functional competence and not the concomitant cessation of proliferation or shift in cell cycle distribution.
在HL-60细胞诱导髓系分化过程中,对编码吞噬细胞细胞色素b重链亚基的X-CGD基因的表达进行了研究。用视黄酸和二甲基甲酰胺联合方案孵育细胞,导致粒细胞形态分化并获得硝基蓝四氮唑还原能力,这是超氧化物生成的一种度量指标。在诱导分化的5天过程中,X-CGD mRNA转录本水平上升了13倍,在接触视黄酸3小时内可检测到2倍的增加。通过核转录分析确定的X-CGD基因相对转录率,在诱导分化24至72小时后增加了2至8倍。然而,X-CGD mRNA水平的变化大于转录率的变化,这意味着也涉及转录后调控。通过离心淘析将细胞周期分为不同阶段,结果显示在诱导前X-CGD转录本主要在G1期细胞中表达,诱导后24小时在细胞周期的所有阶段均有表达。因此,诱导细胞中X-CGD表达的快速增加反映了功能能力的获得,而不是增殖的同时停止或细胞周期分布的改变。