Identification of apoptotic tyrosine-phosphorylated proteins after etoposide or retinoic acid treatment of HL-60 cells
Identification of apoptotic tyrosine-phosphorylated proteins after etoposide or retinoic acid treatment of HL-60 cells
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DOI：
10.1002/pmic.200300671
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发表时间：
2004-04-01
期刊：
影响因子：
3.4
通讯作者：
Magnusson, KE
中科院分区：
文献类型：
--
作者：
Navakauskiene, R;Treigyte, G;Magnusson, KE
使用HL-60细胞作为粒细胞-巨噬细胞分化模型的主要缺点是分化群体中的一些细胞经历凋亡。为了解决这个问题，我们已经确定了酪氨酸磷酸化蛋白分别参与细胞凋亡和分化。HL-60细胞特异性诱导68 μ m依托泊苷进行凋亡，并进行粒细胞分化与1 μ m维甲酸（RA）。比较处理后细胞酪氨酸磷酸化蛋白质的二维电泳图谱。足叶乙甙处理HL-60细胞8h后，83%的细胞发生凋亡。RA处理120 h后，50%的细胞发生凋亡。在依托泊苷处理8小时和RA处理120小时的细胞中均发现了18种胞浆和核酪氨酸磷酸化蛋白，但在增殖的HL-60细胞群中未发现。基质辅助激光解吸/电离飞行时间质谱分析表明，一些蛋白质可能参与信号转导途径（NF κ B、GTP结合蛋白、蛋白质二硫键异构酶、亲环素A），细胞转录和翻译控制中的其他（hnRNP H、hnRNP L、Hsp 60、Hp 1、Hcc-1、26 S蛋白酶体β亚基、ATP合成酶β链），以及细胞细胞骨架组织和受体循环中的第三组（profilin、caveolin-1）。通过筛选酪氨酸磷酸化的凋亡相关蛋白，了解凋亡启动的信号转导机制，可能为白血病的治疗提供新的靶点。
A main shortcoming of using HL-60 cells as a model of granulocyte-macrophage differentiation is that some cells in the differentiating population undergo apoptosis. To address this issue, we have identified which tyrosine-phosphorylated proteins are involved in apoptosis and differentiation, respectively. HL-60 cells were induced specifically to undergo apoptosis with 68 mum etoposide, and to undergo granulocytic differentiation with 1 mum retinoic acid (RA). The corresponding two-dimensional electrophoretic maps of tyrosine-phosphorylated proteins from treated cells were compared. In the 8 h etoposide-treated HL-60 cell population, 83% of the cells were apoptotic. In the 120 h RA-treated cells, 50% of the cells were apoptotic. Eighteen cytosolic and nuclear tyrosine-phosphorylated proteins were found in both the 8 h etoposide- and the 120 h RA-treated cells, but not in the proliferating HL-60 cell population. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry analyses suggested that some of the proteins may be involved in signal transduction pathways (NFkappaB, GTP-binding protein, protein disulfide isomerase, Cyclophilin A), others in cell transcriptional and translational control (hnRNP H, hnRNP L, Hsp60, Hp1, Hcc-1, 26S proteasome beta-subunit, ATP synthase beta-chain), and a third group in cell cytoskeleton organization and receptor cycling (profilin, caveolin-1). An understanding of signal transduction in apoptosis initiation by screening for tyrosine-phosphorylated proteins associated with apoptosis may provide new targets for the treatment of leukemia.