c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells

Abstract The nuclear factor kappa B (NF-κB) signalling pathway plays a crucial role in the regulation of inflammation, and previous research from our lab and others suggests that c-Ski has potential anti-inflammatory effects. However, the role and mechanism of c-Ski, which are related to the regulat...

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Main Authors: Yan Peng, Ren-Ping Xiong, Bo Wang, Xing Chen, Ya-Lie Ning, Yan Zhao, Nan Yang, Jing Zhang, Chang-Hong Li, Yuan-Guo Zhou, Ping Li
Format: Article
Language:English
Published: BMC 2025-04-01
Series:Cell Communication and Signaling
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Online Access:https://doi.org/10.1186/s12964-025-02178-z
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author Yan Peng
Ren-Ping Xiong
Bo Wang
Xing Chen
Ya-Lie Ning
Yan Zhao
Nan Yang
Jing Zhang
Chang-Hong Li
Yuan-Guo Zhou
Ping Li
author_facet Yan Peng
Ren-Ping Xiong
Bo Wang
Xing Chen
Ya-Lie Ning
Yan Zhao
Nan Yang
Jing Zhang
Chang-Hong Li
Yuan-Guo Zhou
Ping Li
author_sort Yan Peng
collection DOAJ
description Abstract The nuclear factor kappa B (NF-κB) signalling pathway plays a crucial role in the regulation of inflammation, and previous research from our lab and others suggests that c-Ski has potential anti-inflammatory effects. However, the role and mechanism of c-Ski, which are related to the regulation of the NF-κB pathway, are still unclear. Here, U937 cells were used, and increasing c-Ski protein levels inhibited inflammatory factor production, invasion, and phagocytosis. The anti-inflammatory effect of c-Ski was similar to that of hormones. Subsequently, immunoprecipitation (IP), Western blot (WB), electrophoretic mobility shift assays (EMSAs), and dual-luciferase reporter assays were used to determine whether increasing c-Ski protein levels could increase c-Ski binding to NF-κB p65 (p65), leading to a decrease in the acetylation level and transcriptional activity of p65. Conversely, decreased p65 expression through targeted small interfering RNA (siRNA) caused the loss of the anti-inflammatory effects of c-Ski. Furthermore, immunoprecipitation confirmed the mutual interaction of c-Ski with HDAC1 and p65, and WB revealed that the anti-inflammatory effect of c-Ski was achieved through the deacetylation of p65 by HDAC1 combined with HDAC1 siRNA and inhibitors. Additionally, through quantitative proteomic analysis, we determined that increasing c-Ski levels had inhibitory effects on the NF-κB pathway. Finally, similar results were also obtained using primary bone marrow-derived macrophages (BMDMs). These findings not only confirm the anti-inflammatory effect of c-Ski but also reveal novel molecular pathways and regulatory molecules of c-Ski, which may be promising targets for direct intervention in the inflammatory response through regulation of c-Ski.
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issn 1478-811X
language English
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publisher BMC
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spelling doaj-art-73d4159a650041638150d63ce6f127132025-08-20T02:25:41ZengBMCCell Communication and Signaling1478-811X2025-04-0123111710.1186/s12964-025-02178-zc-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cellsYan Peng0Ren-Ping Xiong1Bo Wang2Xing Chen3Ya-Lie Ning4Yan Zhao5Nan Yang6Jing Zhang7Chang-Hong Li8Yuan-Guo Zhou9Ping Li10State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)State Key Laboratory of Trauma, Burn and Combined Injury, Department of Army Occupational Disease, Daping Hospital, Army Medical University (Third Military Medical University)Abstract The nuclear factor kappa B (NF-κB) signalling pathway plays a crucial role in the regulation of inflammation, and previous research from our lab and others suggests that c-Ski has potential anti-inflammatory effects. However, the role and mechanism of c-Ski, which are related to the regulation of the NF-κB pathway, are still unclear. Here, U937 cells were used, and increasing c-Ski protein levels inhibited inflammatory factor production, invasion, and phagocytosis. The anti-inflammatory effect of c-Ski was similar to that of hormones. Subsequently, immunoprecipitation (IP), Western blot (WB), electrophoretic mobility shift assays (EMSAs), and dual-luciferase reporter assays were used to determine whether increasing c-Ski protein levels could increase c-Ski binding to NF-κB p65 (p65), leading to a decrease in the acetylation level and transcriptional activity of p65. Conversely, decreased p65 expression through targeted small interfering RNA (siRNA) caused the loss of the anti-inflammatory effects of c-Ski. Furthermore, immunoprecipitation confirmed the mutual interaction of c-Ski with HDAC1 and p65, and WB revealed that the anti-inflammatory effect of c-Ski was achieved through the deacetylation of p65 by HDAC1 combined with HDAC1 siRNA and inhibitors. Additionally, through quantitative proteomic analysis, we determined that increasing c-Ski levels had inhibitory effects on the NF-κB pathway. Finally, similar results were also obtained using primary bone marrow-derived macrophages (BMDMs). These findings not only confirm the anti-inflammatory effect of c-Ski but also reveal novel molecular pathways and regulatory molecules of c-Ski, which may be promising targets for direct intervention in the inflammatory response through regulation of c-Ski.https://doi.org/10.1186/s12964-025-02178-zc-SkiNF-κBInflammationp65HDAC1
spellingShingle Yan Peng
Ren-Ping Xiong
Bo Wang
Xing Chen
Ya-Lie Ning
Yan Zhao
Nan Yang
Jing Zhang
Chang-Hong Li
Yuan-Guo Zhou
Ping Li
c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
Cell Communication and Signaling
c-Ski
NF-κB
Inflammation
p65
HDAC1
title c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
title_full c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
title_fullStr c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
title_full_unstemmed c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
title_short c-Ski is a novel repressor of NF-κB through interaction with p65 and HDAC1 in U937 cells
title_sort c ski is a novel repressor of nf κb through interaction with p65 and hdac1 in u937 cells
topic c-Ski
NF-κB
Inflammation
p65
HDAC1
url https://doi.org/10.1186/s12964-025-02178-z
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