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Inflammation Research
Published in: Inflammation Research 8/2012

01-08-2012 | Original Research Paper

AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells

Authors: Wenying Zou, Xinwei Chu, Chunqing Cai, Mengchen Zou, Xiaojing Meng, Haiyang Chen, Fei Zou

Published in: Inflammation Research | Issue 8/2012

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Abstract

Objectives

Neutrophil 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.

Methods

HL-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.

Results

We 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.

Conclusion

This study demonstrates that AKT activation plays a crucial role in dHL-60 cell polarization.
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Metadata
Title
AKT-mediated regulation of polarization in differentiated human neutrophil-like HL-60 cells
Authors
Wenying Zou
Xinwei Chu
Chunqing Cai
Mengchen Zou
Xiaojing Meng
Haiyang Chen
Fei Zou
Publication date
01-08-2012
Publisher
SP Birkhäuser Verlag Basel
Published in
Inflammation Research / Issue 8/2012
Print ISSN: 1023-3830
Electronic ISSN: 1420-908X
DOI
https://doi.org/10.1007/s00011-012-0478-y

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