Inhibitor of Tec kinase, LFM-A13, decreases pro-inflammatory mediators production in LPS-stimulated RAW264.7 macrophages via NF-κB pathway.

Fei Wang,Hao Cheng,Xu-Lin Chen,Chao Wang,Wei Zhang,Xu Fang,Sheng Liu

doi:10.18632/oncotarget.16212

Fei Wang, Hao Cheng + Show 5 more

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https://doi.org/10.18632/oncotarget.16212

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Abstract

Tec kinase, a prototypical member of the Tec tyrosine kinases family, was shown to mainly govern lymphocyte proliferation. In the present study, we investigated the role of Tec kinase in acute inflammatory response in lipopolysaccharide (LPS) challenge. First, we demonstrate that Tec kinase activity was observed in RAW264.7 macrophages exposed to LPS. Tec and phosphorylated Tec expression were upregulated in a dose- and time-dependent manner after LPS stimulation. LPS increased monocyte chemotactic protein (MCP)-1 secretion and intercellular adhesion molecule (ICAM)-1 expression, and increasing mRNA expression was consistently observed. LPS also induced IκBα phoshporylaytion and its degradation, increased NF-κB p65 phoshporylaytion and translocation to nuclei in RAW264.7 cells. Pretreatment with LFM-A13 decreased LPS-induced cytokines and chemokines production and mRNA levels, blocked NF-κB transactivation. These effects of LPS were also prevented by Tec-siRNA. Additionally, LFM-A13 or Tec-siRNA obviously inhibited LPS-induced TGFβ-activated kinase 1(TAK1) phosphorylation. Taken together, our results suggest that Tec kinase involves in acute inflammation process in LPS-stimulated RAW264.7 cells, at least mediated by activating TAK1/ NF-κB signal pathway.

Highlights

Innate immune cells such as macrophages and neutrophils constitute a front line of defense against most microbial infection and are responsible for invading pathogens
We examined whether the Tec kinase was activated after LPS stimulation in macrophages
Tec family kinases are recruited to the plasma membrane through binging of their pleckstrin homology (PH) domain to phosphatidylinositol(3,4,5)trisphosphated (PIP3) [14]

Summary

Introduction

Innate immune cells such as macrophages and neutrophils constitute a front line of defense against most microbial infection and are responsible for invading pathogens. Non-receptor tyrosine kinase family, consist of five members: Tec, Btk, ITK/Emt/Tsk, Bmx and Txk/Rlk [3]. They constitute the second largest family of cytoplasmic tyrosine kinases in humans. Tec kinase is suppressed by Insulinlike growth factor (IGF)-1 and augments cell survival by inhibition of necrosis [7]. Differentiation and cell survival, Tec kinase families are involved in the regulation of cytokines expression. Tec and Btk kinases were shown to be expressed and functional, activated by the chemoattractant fMLP [8]. Gilbert C et al showed that Tec kinases increased fMLP-induced superoxide production, adhesion, and chemotaxis through p38 MAPK in human neutrophils [9]

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