Murphy J J, Norton J D
Department of Haematology, Royal Free Hospital School of Medicine, London, U.K.
Biochim Biophys Acta. 1990 Jul 30;1049(3):261-71. doi: 10.1016/0167-4781(90)90096-k.
Monoclonal B lymphocytes from B cell chronic lymphocytic leukemia (B-CLL) can be induced to undergo plasmacytoid differentiation in vitro by the phorbol ester, phorbol 12-myristate 13-acetate (PMA). By differential screening of a cDNA library derived from cells treated with phorbol ester we have isolated and characterised a set of early response genes (ERGs) displaying rapid transient up-regulation of expression in response to PMA. Cross-hybridisation studies showed that PMA probably induces the expression of over one hundred distinct genes, implying an ERG complexity comparable to that activated by mitogenic stimulation of fibroblasts and normal T lymphocytes. Of 13 genes analysed in detail, most were induced by PMA without a requirement for de novo protein synthesis, whilst nuclear run-on analysis showed that at least some of the more abundant classes of ERG were up-regulated through transcriptional mechanisms. In a proliferating variant B-CLL population, few differences in ERG expression were seen, suggesting that these genes are part of a gene regulatory pathway coupled to the differentiative rather than the proliferative response of B-CLL cells. However, studies in a range of cell types revealed a surprisingly diverse pattern of PMA-induced expression where most ERGs were relatively B-CLL-specific. This implies an extreme diversity of gene regulatory pathways activated in the primary response by phorbol ester generally and suggests that the onset of PMA-induced plasmacytoid differentiation of B-CLL cells is preceded by activation of a complex gene regulatory program that is largely unique to this maturation-arrested B cell.
来自B细胞慢性淋巴细胞白血病(B-CLL)的单克隆B淋巴细胞可被佛波酯——佛波醇12-肉豆蔻酸酯13-乙酸酯(PMA)在体外诱导发生浆细胞样分化。通过对来自经佛波酯处理的细胞的cDNA文库进行差异筛选,我们分离并鉴定了一组早期反应基因(ERGs),这些基因在对PMA的反应中表现出表达的快速瞬时上调。交叉杂交研究表明,PMA可能诱导了一百多个不同基因的表达,这意味着ERG的复杂性与成纤维细胞和正常T淋巴细胞的有丝分裂刺激所激活的复杂性相当。在详细分析的13个基因中,大多数基因由PMA诱导,无需从头合成蛋白质,而细胞核连续分析表明,至少一些较丰富类别的ERG是通过转录机制上调的。在一个增殖性变异的B-CLL群体中,未观察到ERG表达有明显差异,这表明这些基因是与B-CLL细胞的分化反应而非增殖反应相关的基因调控途径的一部分。然而,对一系列细胞类型的研究揭示了PMA诱导表达的惊人多样模式,其中大多数ERG相对具有B-CLL特异性。这意味着佛波酯在初级反应中激活的基因调控途径极其多样,并且表明在PMA诱导B-CLL细胞发生浆细胞样分化之前,先激活了一个复杂的基因调控程序,该程序在很大程度上是这种成熟停滞的B细胞所特有的。