LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ

Abstract Background Neutrophil extracellular traps (NETs) are pivotal in the metastasis of non‐small cell lung cancer (NSCLC). Our previous research demonstrated that NETs facilitate NSCLC metastasis by triggering the stimulation of the NOD‐like receptor protein 3 (NLRP3) inflammasome, which is medi...

Full description

Saved in:
Bibliographic Details
Main Authors: Xin Ye, Chen Fang, Weiwei Hong, Xiaoying Qian, Biao Yu, Bingbiao Zhou, Xinyuan Yao, Dengying Chen, Chengsi Shu, Chuanhong Luo, Yong Wang, Yong Li
Format: Article
Language:English
Published: Wiley 2025-06-01
Series:Clinical and Translational Medicine
Subjects:
Online Access:https://doi.org/10.1002/ctm2.70342
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850168855559667712
author Xin Ye
Chen Fang
Weiwei Hong
Xiaoying Qian
Biao Yu
Bingbiao Zhou
Xinyuan Yao
Dengying Chen
Chengsi Shu
Chuanhong Luo
Yong Wang
Yong Li
author_facet Xin Ye
Chen Fang
Weiwei Hong
Xiaoying Qian
Biao Yu
Bingbiao Zhou
Xinyuan Yao
Dengying Chen
Chengsi Shu
Chuanhong Luo
Yong Wang
Yong Li
author_sort Xin Ye
collection DOAJ
description Abstract Background Neutrophil extracellular traps (NETs) are pivotal in the metastasis of non‐small cell lung cancer (NSCLC). Our previous research demonstrated that NETs facilitate NSCLC metastasis by triggering the stimulation of the NOD‐like receptor protein 3 (NLRP3) inflammasome, which is mediated through the suppression of the long non‐coding RNA MIR503HG. However, the precise molecular mechanisms linking MIR503HG to NLRP3 are still not fully understood. Methods By employing protein mass spectrometry and the Human TFDB database, key molecules involved in NLRP3 regulation were identified. The involvement of CCAAT enhancer binding protein beta (C/EBPβ) in NSCLC metastasis was examined in both cellular and animal models. Dual‐luciferase and CUT&RUN assays confirmed the mechanism by which C/EBPβ controls NLRP3. The regulatory relationship between MIR503HG and C/EBPβ was explored through RNA pulldown, RNA immunoprecipitation and coimmunoprecipitation assays. Additionally, methylation‐specific PCR and other studies revealed that NETs suppress MIR503HG via DNA methylation. Results We found that C/EBPβ mediates the regulation of NLRP3 by MIR503HG. Further investigation confirmed that C/EBPβ promotes the migration and invasion of NSCLC both in vivo and in vitro and is highly expressed in NSCLC tissue. Mechanistically, C/EBPβ binds to the NLRP3 promoter to promote NLRP3 expression. Conversely, MIR503HG suppressed C/EBPβ expression by facilitating C/EBPβ interaction with the E3 ubiquitin ligase RNF43, which in turn reduced NLRP3 expression and NSCLC metastasis. Meanwhile, we investigated the mechanism by which NETs inhibit MIR503HG expression and found that DNA methylation is involved in the suppression of MIR503HG by NETs. Additionally, reversing this methylation partially restored MIR503HG and NLRP3 expression and mitigated the metastatic effects of NETs in NSCLC. Conclusions This study emphasises the critical roles of C/EBPβ and DNA methylation in NETs‐mediated NSCLC metastasis. These findings unveil C/EBPβ and DNA methylation as potential novel targets for NSCLC with high NETs expression. Key points NETs suppress the expression of MIR503HG by inducing promoter DNA methylation. C/EBPβ binds to the NLRP3 promoter to promote NLRP3 expression. MIR503HG inhibits the expression of C/EBPβ protein by promoting the interaction between C/EBPβ and the E3 ubiquitin ligase RNF43, thereby repressing NLRP3 expression.
format Article
id doaj-art-1e35641d30f04ddc8d5ee9c1057a8404
institution OA Journals
issn 2001-1326
language English
publishDate 2025-06-01
publisher Wiley
record_format Article
series Clinical and Translational Medicine
spelling doaj-art-1e35641d30f04ddc8d5ee9c1057a84042025-08-20T02:20:52ZengWileyClinical and Translational Medicine2001-13262025-06-01156n/an/a10.1002/ctm2.70342LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβXin Ye0Chen Fang1Weiwei Hong2Xiaoying Qian3Biao Yu4Bingbiao Zhou5Xinyuan Yao6Dengying Chen7Chengsi Shu8Chuanhong Luo9Yong Wang10Yong Li11Department of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaDepartment of Medical Oncology The First Affiliated Hospital of Nanchang University Nanchang ChinaAbstract Background Neutrophil extracellular traps (NETs) are pivotal in the metastasis of non‐small cell lung cancer (NSCLC). Our previous research demonstrated that NETs facilitate NSCLC metastasis by triggering the stimulation of the NOD‐like receptor protein 3 (NLRP3) inflammasome, which is mediated through the suppression of the long non‐coding RNA MIR503HG. However, the precise molecular mechanisms linking MIR503HG to NLRP3 are still not fully understood. Methods By employing protein mass spectrometry and the Human TFDB database, key molecules involved in NLRP3 regulation were identified. The involvement of CCAAT enhancer binding protein beta (C/EBPβ) in NSCLC metastasis was examined in both cellular and animal models. Dual‐luciferase and CUT&RUN assays confirmed the mechanism by which C/EBPβ controls NLRP3. The regulatory relationship between MIR503HG and C/EBPβ was explored through RNA pulldown, RNA immunoprecipitation and coimmunoprecipitation assays. Additionally, methylation‐specific PCR and other studies revealed that NETs suppress MIR503HG via DNA methylation. Results We found that C/EBPβ mediates the regulation of NLRP3 by MIR503HG. Further investigation confirmed that C/EBPβ promotes the migration and invasion of NSCLC both in vivo and in vitro and is highly expressed in NSCLC tissue. Mechanistically, C/EBPβ binds to the NLRP3 promoter to promote NLRP3 expression. Conversely, MIR503HG suppressed C/EBPβ expression by facilitating C/EBPβ interaction with the E3 ubiquitin ligase RNF43, which in turn reduced NLRP3 expression and NSCLC metastasis. Meanwhile, we investigated the mechanism by which NETs inhibit MIR503HG expression and found that DNA methylation is involved in the suppression of MIR503HG by NETs. Additionally, reversing this methylation partially restored MIR503HG and NLRP3 expression and mitigated the metastatic effects of NETs in NSCLC. Conclusions This study emphasises the critical roles of C/EBPβ and DNA methylation in NETs‐mediated NSCLC metastasis. These findings unveil C/EBPβ and DNA methylation as potential novel targets for NSCLC with high NETs expression. Key points NETs suppress the expression of MIR503HG by inducing promoter DNA methylation. C/EBPβ binds to the NLRP3 promoter to promote NLRP3 expression. MIR503HG inhibits the expression of C/EBPβ protein by promoting the interaction between C/EBPβ and the E3 ubiquitin ligase RNF43, thereby repressing NLRP3 expression.https://doi.org/10.1002/ctm2.70342C/EBPβDNA methylationMIR503HGNETsNLRP3RNF43
spellingShingle Xin Ye
Chen Fang
Weiwei Hong
Xiaoying Qian
Biao Yu
Bingbiao Zhou
Xinyuan Yao
Dengying Chen
Chengsi Shu
Chuanhong Luo
Yong Wang
Yong Li
LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
Clinical and Translational Medicine
C/EBPβ
DNA methylation
MIR503HG
NETs
NLRP3
RNF43
title LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
title_full LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
title_fullStr LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
title_full_unstemmed LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
title_short LncRNA MIR503HG regulates NETs‐mediated NLRP3 inflammasome activation and NSCLC metastasis by enhancing the ubiquitination of C/EBPβ
title_sort lncrna mir503hg regulates nets mediated nlrp3 inflammasome activation and nsclc metastasis by enhancing the ubiquitination of c ebpβ
topic C/EBPβ
DNA methylation
MIR503HG
NETs
NLRP3
RNF43
url https://doi.org/10.1002/ctm2.70342
work_keys_str_mv AT xinye lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT chenfang lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT weiweihong lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT xiaoyingqian lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT biaoyu lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT bingbiaozhou lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT xinyuanyao lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT dengyingchen lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT chengsishu lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT chuanhongluo lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT yongwang lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb
AT yongli lncrnamir503hgregulatesnetsmediatednlrp3inflammasomeactivationandnsclcmetastasisbyenhancingtheubiquitinationofcebpb