Yousefi S, Green D R, Blaser K, Simon H U
Swiss Institute of Allergy and Asthma Research, Davos.
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10868-72. doi: 10.1073/pnas.91.23.10868.
Early signaling events that control the process of programmed cell death are largely unknown. Tyrosine phosphorylation plays a major role in transmembrane signal transduction through most cell surface receptors. Granulocyte/macrophage colony-stimulating factor (GM-CSF), a cytokine released by activated T cells, has been shown to increase tyrosine phosphorylation in several cells and to inhibit granulocyte cell death in vitro. In this study, we demonstrate that the effect of GM-CSF on granulocyte cell death can be blocked by the tyrosine kinase inhibitor genistein, suggesting that increases in tyrosine phosphorylation are essential to inhibit cell death. To analyze the role of tyrosine phosphorylation for the regulation of granulocyte cell death more precisely, we increased levels of tyrosine phosphorylation using the protein-tyrosine phosphatase inhibitor phenylarsine oxide (PAO). Similar to GM-CSF, treatment of the cells with PAO was followed by high increases in tyrosine phosphorylation and inhibition of programmed cell death in human eosinophils and neutrophils. Strikingly, at low concentrations of the inhibitor and low induction of tyrosine phosphorylation, acceleration of apoptosis was observed. Genistein and herbimycin A reversed the effects of PAO on tyrosine phosphorylation and granulocyte apoptosis. These results suggest that programmed eosinophil and neutrophil death is regulated by early events of signal transduction pathways such as tyrosine phosphorylation.
控制程序性细胞死亡过程的早期信号事件在很大程度上尚不明确。酪氨酸磷酸化在通过大多数细胞表面受体进行的跨膜信号转导中起主要作用。粒细胞/巨噬细胞集落刺激因子(GM-CSF)是活化T细胞释放的一种细胞因子,已被证明可增加多种细胞中的酪氨酸磷酸化,并在体外抑制粒细胞死亡。在本研究中,我们证明GM-CSF对粒细胞死亡的作用可被酪氨酸激酶抑制剂染料木黄酮阻断,这表明酪氨酸磷酸化的增加对于抑制细胞死亡至关重要。为了更精确地分析酪氨酸磷酸化在调节粒细胞死亡中的作用,我们使用蛋白酪氨酸磷酸酶抑制剂氧化苯胂(PAO)提高酪氨酸磷酸化水平。与GM-CSF相似,用PAO处理细胞后,人嗜酸性粒细胞和中性粒细胞中的酪氨酸磷酸化大幅增加,程序性细胞死亡受到抑制。引人注目的是,在低浓度抑制剂和低酪氨酸磷酸化诱导情况下,观察到细胞凋亡加速。染料木黄酮和赫曲霉素A可逆转PAO对酪氨酸磷酸化和粒细胞凋亡的影响。这些结果表明,嗜酸性粒细胞和中性粒细胞的程序性死亡受信号转导途径的早期事件如酪氨酸磷酸化的调节。