Clara cell 10-kDa protein gene transfection inhibits NF-κB activity in airway epithelial cells.

PloS One
Xiao-Bo LongZheng Liu

Abstract

Clara cell 10-kDa protein (CC10) is a multifunctional protein with anti-inflammatory and immunomodulatory effects. Induction of CC10 expression by gene transfection may possess potential therapeutic effect. Nuclear factor κB (NF-κB) plays a key role in the inflammatory processes of airway diseases. To investigate potential therapeutic effect of CC10 gene transfection in controlling airway inflammation and the underlying intracellular mechanisms, in this study, we constructed CC10 plasmid and transfected it into bronchial epithelial cell line BEAS-2B cells and CC10 knockout mice. In BEAS-2B cells, CC10's effect on interleukin (IL)-1β induced IL-8 expression was explored by means of RT-PCR and ELISA and its effect on NF-κB classical signaling pathway was studied by luciferase reporter, western blot, and immunoprecipitation assay. The effect of endogenous CC10 on IL-1β evoked IL-8 expression was studied by means of nasal explant culture. In mice, CC10's effect on IL-1β induced IL-8 and nuclear p65 expression was examined by immunohistochemistry. First, we found that the CC10 gene transfer could inhibit IL-1β induced IL-8 expression in BEAS-2B cells. Furthermore, we found that CC10 repressed IL-1β induced NF-κB activation by inhibi...Continue Reading

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Citations

Dec 18, 2012·Current Opinion in Allergy and Clinical Immunology·Heng WangZheng Liu
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Jan 16, 2021·American Journal of Respiratory and Critical Care Medicine·Hong Wei Chu, Niccolette Schaunaman

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Methods Mentioned

BETA
transfection
PCR
ELISA
Assay
electrophoresis
Co-Immunoprecipitation
nuclear translocation
immunoprecipitation
biopsy
coimmunoprecipitation

Software Mentioned

IS1000
Pro Plus
Image

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