AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells
AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells
复制标题
AKT 介导的分化人中性粒细胞样 HL-60 细胞极化调节
DOI：
10.1007/s00011-012-0478-y
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发表时间：
2012-05
影响因子：
6.7
通讯作者：
Zou, Fei
中科院分区：
文献类型：
--
作者：
Zou, Wenying;Chu, Xinwei;Cai, Chunqing;Zou, Mengchen;Meng, Xiaojing;Chen, Haiyang;Zou, Fei
目的神经元极化是炎症反应的关键。AKT是一种丝氨酸/苏氨酸蛋白激酶，与细胞迁移有关。然而，AKT是否影响中性粒细胞极化尚不完全清楚。在这项研究中，我们测试的假设，AKT调节极化的嗜中性粒细胞样分化的HL-60细胞（dHL-60）在fMLP.MethodsHL-60细胞分化成dHL-60通过培养在培养基中含有1.3%DMSO长达6天。用Zigmond小室测定dHL-60细胞和原代人中性粒细胞的极化。通过免疫荧光和Western印迹分析磷酸化Akt。用罗丹明-鬼笔定染色法检测F-肌动蛋白聚合。Rac 2激活使用GST下拉assay.ResultsWe发现，细胞极化率的变化是一致的AKT磷酸化水平的变化在HL-60细胞分化过程中响应fMLP。此外，在用PI 3激酶抑制剂或AKT抑制剂预处理的fMLP刺激的dHL-60细胞中，细胞极化和AKT磷酸化减少，这在原代人中性粒细胞中得到证实。AKT抑制剂改变fMLP诱导的F-肌动蛋白聚合。Rac 2 GTPases也下降的AKT抑制剂在fMLP刺激的dHL-60 cells.ConclusionThis study demonstrates，AKT激活dHL-60细胞极化中起着至关重要的作用。
ObjectivesNeutrophil polarization is critical for the inflammatory response. AKT is a serine/threonine protein kinase and has been implicated in cell migration. However, it is not completely clear whether AKT affects neutrophil polarization. In this study, we tested the hypothesis that AKT regulates the polarization of neutrophil-like differentiated HL-60 cells (dHL-60) in response to fMLP.MethodsHL-60 cells were differentiated into dHL-60 by incubation in medium containing 1.3 % DMSO for up to 6 days. Polarization of dHL-60 cells and primary human neutrophils were measured by Zigmond chamber. Phospho-Akt was analyzed by immunofluorescence and Western blot analysis. F-actin polymerization was detected by Rhodamine-Phalloidine staining. Rac2 activation was evaluated using GST Pull-down assay.ResultsWe found that changes in the rate of cell polarization were consistent with the changes in AKT phosphorylation levels during HL-60 cell differentiation in response to fMLP. Moreover, cell polarization and AKT phosphorylation were reduced in fMLP-stimulated dHL-60 cells pretreated with the PI3 kinase inhibitors or the AKT inhibitors, which was confirmed in the primary human neutrophils. The AKT inhibitors altered fMLP-induced F-actin polymerization. Rac2 GTPases was also decreased by the AKT inhibitors in fMLP-stimulated dHL-60 cells.ConclusionThis study demonstrates that AKT activation plays a crucial role in dHL-60 cell polarization.
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DOI：
10.1126/science.1170179
发表时间：
2009-04-17
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Nishikimi A;Fukuhara H;Su W;Hongu T;Takasuga S;Mihara H;Cao Q;Sanematsu F;Kanai M;Hasegawa H;Tanaka Y;Shibasaki M;Kanaho Y;Sasaki T;Frohman MA;Fukui Y
通讯作者：
Fukui Y
影响因子：
4
作者：
D. McKay;J. Kusel;P. Wilkinson
通讯作者：
D. McKay;J. Kusel;P. Wilkinson
影响因子：
4.4
作者：
Boulven, Isaline;Levasseur, Sylvain;Naccache, Paul H.
通讯作者：
Naccache, Paul H.
影响因子：
64.5
作者：
RIDLEY, AJ;PATERSON, HF;HALL, A
通讯作者：
HALL, A
影响因子：
3.3
作者：
Lee, S;Comer, FI;Firtel, RA
通讯作者：
Firtel, RA