The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma

BackgroundLung adenocarcinoma (LUAD) is a major subtype of lung cancer and one of the deadliest cancers in humans. Dysregulation of miRNA activity in tumor-associated neutrophils (TANs) in the tumor microenvironment plays an important role in the occurrence and development of LUAD.MethodIn this stud...

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Main Authors: Xiaojing Zhang, Xitong Huang, Xianying Zhang, Lichang Lai, Baoyi Zhu, Peibin Lin, Zhanfang Kang, Dazhong Yin, Dongbo Tian, Zisheng Chen, Jun Gao
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1561081/full
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author Xiaojing Zhang
Xitong Huang
Xianying Zhang
Lichang Lai
Baoyi Zhu
Peibin Lin
Zhanfang Kang
Dazhong Yin
Dongbo Tian
Zisheng Chen
Jun Gao
author_facet Xiaojing Zhang
Xitong Huang
Xianying Zhang
Lichang Lai
Baoyi Zhu
Peibin Lin
Zhanfang Kang
Dazhong Yin
Dongbo Tian
Zisheng Chen
Jun Gao
author_sort Xiaojing Zhang
collection DOAJ
description BackgroundLung adenocarcinoma (LUAD) is a major subtype of lung cancer and one of the deadliest cancers in humans. Dysregulation of miRNA activity in tumor-associated neutrophils (TANs) in the tumor microenvironment plays an important role in the occurrence and development of LUAD.MethodIn this study, the miReact algorithm was used to analyze the single-cell RNA sequencing data of LUAD samples to reveal the miRNA profile characteristics of TANs in LUAD patients. The function of miR-941 was investigated in vivo and in vitro. The target gene and underlying signaling pathway of miR-941 were predicted and validated with qPCR, luciferase assay, WB and ELISA assay.ResultsThe results indicated the crucial role of TANs, especially N2-TANs in LUAD and miR-941 activity was significantly upregulated in TANs of LUAD patients. MiR-941 overexpression promoted the proliferation, invasion, migration and anti-apoptosis of A549 and H1299. In vivo xenograft mouse model confirmed that miR-941 overexpression enhanced the growth of tumors formed by H1299 cells. Bioinformatics analysis showed that miR-941 targeted the tumor suppressor gene FOXN4, and we confirmed that FOXN4 overexpression could counteract the malignant effects of miR-941. In addition, miR-941 may drive LUAD progression through the FOXN4/TGF-β feedback signaling loop and participate in the N2-TAN polarization.ConclusionIn summary, these findings reveal the key role of N2-TANs and the miR-941/FOXN4/TGF-β signaling loop in LUAD progression and provide potential therapeutic targets for future interventions.
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spelling doaj-art-e2ad70722b4044dab8a1d523a66357202025-08-20T03:14:17ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-04-011610.3389/fimmu.2025.15610811561081The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinomaXiaojing Zhang0Xitong Huang1Xianying Zhang2Lichang Lai3Baoyi Zhu4Peibin Lin5Zhanfang Kang6Dazhong Yin7Dongbo Tian8Zisheng Chen9Jun Gao10Department of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Traditional Chinese Medicine, China Pharmaceutical University, Nanjing, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaDepartment of Respiratory and Critical Care Medicine, The Affiliated Qingyuan Hospital (Qingyuan People’s Hospital), Guangzhou Medical University, Qingyuan, ChinaBackgroundLung adenocarcinoma (LUAD) is a major subtype of lung cancer and one of the deadliest cancers in humans. Dysregulation of miRNA activity in tumor-associated neutrophils (TANs) in the tumor microenvironment plays an important role in the occurrence and development of LUAD.MethodIn this study, the miReact algorithm was used to analyze the single-cell RNA sequencing data of LUAD samples to reveal the miRNA profile characteristics of TANs in LUAD patients. The function of miR-941 was investigated in vivo and in vitro. The target gene and underlying signaling pathway of miR-941 were predicted and validated with qPCR, luciferase assay, WB and ELISA assay.ResultsThe results indicated the crucial role of TANs, especially N2-TANs in LUAD and miR-941 activity was significantly upregulated in TANs of LUAD patients. MiR-941 overexpression promoted the proliferation, invasion, migration and anti-apoptosis of A549 and H1299. In vivo xenograft mouse model confirmed that miR-941 overexpression enhanced the growth of tumors formed by H1299 cells. Bioinformatics analysis showed that miR-941 targeted the tumor suppressor gene FOXN4, and we confirmed that FOXN4 overexpression could counteract the malignant effects of miR-941. In addition, miR-941 may drive LUAD progression through the FOXN4/TGF-β feedback signaling loop and participate in the N2-TAN polarization.ConclusionIn summary, these findings reveal the key role of N2-TANs and the miR-941/FOXN4/TGF-β signaling loop in LUAD progression and provide potential therapeutic targets for future interventions.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1561081/fulllung adenocarcinoma (LUAD)single-cell RNA sequencing (scRNA-seq)tumor-associated neutrophils (TANs)miR-941Foxn4TGF-b
spellingShingle Xiaojing Zhang
Xitong Huang
Xianying Zhang
Lichang Lai
Baoyi Zhu
Peibin Lin
Zhanfang Kang
Dazhong Yin
Dongbo Tian
Zisheng Chen
Jun Gao
The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
Frontiers in Immunology
lung adenocarcinoma (LUAD)
single-cell RNA sequencing (scRNA-seq)
tumor-associated neutrophils (TANs)
miR-941
Foxn4
TGF-b
title The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
title_full The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
title_fullStr The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
title_full_unstemmed The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
title_short The miR-941/FOXN4/TGF-β feedback loop induces N2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
title_sort mir 941 foxn4 tgf β feedback loop induces n2 polarization of neutrophils and enhances tumor progression of lung adenocarcinoma
topic lung adenocarcinoma (LUAD)
single-cell RNA sequencing (scRNA-seq)
tumor-associated neutrophils (TANs)
miR-941
Foxn4
TGF-b
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1561081/full
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