NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer

Abstract Cellular glutathione (GSH) in lung cancer cells represents the most abundant antioxidant. GSH production is regulated not only by upregulated cystine/glutamate exchanger (xCT) but also by the involvement of glutamate transporters, specifically excitatory amino acid transporter 3 (EAAT3). Ou...

Full description

Saved in:
Bibliographic Details
Main Authors: Donghua Wen, Wenjing Li, Xiang Song, Min Hu, Yueling Liao, Dongliang Xu, Jiong Deng, Wenzheng Guo
Format: Article
Language:English
Published: Nature Publishing Group 2025-02-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07453-y
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849765378923692032
author Donghua Wen
Wenjing Li
Xiang Song
Min Hu
Yueling Liao
Dongliang Xu
Jiong Deng
Wenzheng Guo
author_facet Donghua Wen
Wenjing Li
Xiang Song
Min Hu
Yueling Liao
Dongliang Xu
Jiong Deng
Wenzheng Guo
author_sort Donghua Wen
collection DOAJ
description Abstract Cellular glutathione (GSH) in lung cancer cells represents the most abundant antioxidant. GSH production is regulated not only by upregulated cystine/glutamate exchanger (xCT) but also by the involvement of glutamate transporters, specifically excitatory amino acid transporter 3 (EAAT3). Our prior research established that the uptake of glutamate via EAAT3 plays a pivotal role in driving cystine uptake through xCT, contributing to GSH biosynthesis during lung tumorigenesis. Nevertheless, the underlying mechanism governing the upregulation of EAAT3 remains enigmatic. In this study, we conducted a comprehensive reanalysis of publicly available data and employed the Gprc5a–/–/SR-IκB mouse model alongside in vitro cell experiments to elucidate the correlations between NF-κB and EAAT3 in lung cancer. We observed that EAAT3 knockdown, similar to NF-κB inhibition, led to the accumulation of reactive oxygen species (ROS) and increased sensitivity to ferroptosis induction by RAS-selective lethal 3 (RSL3). Mechanistic insights were obtained through chromatin immunoprecipitation and luciferase reporter assays, revealing that NF-κB induces EAAT3 expression via two putative cis-elements within its promoter. Furthermore, our investigation unveiled the upregulation of EAAT3 in a subset of clinical non-small cell lung cancer (NSCLC) tissues, exhibiting a positive correlation with the P65 protein. In addition, the inflammatory factor of smoking was found to augment EAAT3 expression in both human and murine experimental models. These findings collectively emphasize the pivotal role of the NF-κB/EAAT3 axis in managing antioxidant stress and influencing lung cancer development. Moreover, this research offers insights into the potential for a combined ferroptosis therapy strategy in lung cancer treatment.
format Article
id doaj-art-e93289ca2bb4410c864db18e0a9e531f
institution DOAJ
issn 2041-4889
language English
publishDate 2025-02-01
publisher Nature Publishing Group
record_format Article
series Cell Death and Disease
spelling doaj-art-e93289ca2bb4410c864db18e0a9e531f2025-08-20T03:04:53ZengNature Publishing GroupCell Death and Disease2041-48892025-02-0116111410.1038/s41419-025-07453-yNF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancerDonghua Wen0Wenjing Li1Xiang Song2Min Hu3Yueling Liao4Dongliang Xu5Jiong Deng6Wenzheng Guo7Department of Laboratory Medicine, Shanghai East Hospital, Tongji University School of MedicineDepartment of Laboratory Medicine, Shanghai East Hospital, Tongji University School of MedicineBreast Cancer Center, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical SciencesDepartment of Laboratory Medicine, Shanghai East Hospital, Tongji University School of MedicineCollege of Life and Environmental Science, Wenzhou UniversityRenji Hospital, Shanghai Jiao Tong University School of MedicineMedical Research Center, Affiliated Hospital of Binzhou Medical UniversityDepartment of Laboratory Medicine, Shanghai East Hospital, Tongji University School of MedicineAbstract Cellular glutathione (GSH) in lung cancer cells represents the most abundant antioxidant. GSH production is regulated not only by upregulated cystine/glutamate exchanger (xCT) but also by the involvement of glutamate transporters, specifically excitatory amino acid transporter 3 (EAAT3). Our prior research established that the uptake of glutamate via EAAT3 plays a pivotal role in driving cystine uptake through xCT, contributing to GSH biosynthesis during lung tumorigenesis. Nevertheless, the underlying mechanism governing the upregulation of EAAT3 remains enigmatic. In this study, we conducted a comprehensive reanalysis of publicly available data and employed the Gprc5a–/–/SR-IκB mouse model alongside in vitro cell experiments to elucidate the correlations between NF-κB and EAAT3 in lung cancer. We observed that EAAT3 knockdown, similar to NF-κB inhibition, led to the accumulation of reactive oxygen species (ROS) and increased sensitivity to ferroptosis induction by RAS-selective lethal 3 (RSL3). Mechanistic insights were obtained through chromatin immunoprecipitation and luciferase reporter assays, revealing that NF-κB induces EAAT3 expression via two putative cis-elements within its promoter. Furthermore, our investigation unveiled the upregulation of EAAT3 in a subset of clinical non-small cell lung cancer (NSCLC) tissues, exhibiting a positive correlation with the P65 protein. In addition, the inflammatory factor of smoking was found to augment EAAT3 expression in both human and murine experimental models. These findings collectively emphasize the pivotal role of the NF-κB/EAAT3 axis in managing antioxidant stress and influencing lung cancer development. Moreover, this research offers insights into the potential for a combined ferroptosis therapy strategy in lung cancer treatment.https://doi.org/10.1038/s41419-025-07453-y
spellingShingle Donghua Wen
Wenjing Li
Xiang Song
Min Hu
Yueling Liao
Dongliang Xu
Jiong Deng
Wenzheng Guo
NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
Cell Death and Disease
title NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
title_full NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
title_fullStr NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
title_full_unstemmed NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
title_short NF-κB-mediated EAAT3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
title_sort nf κb mediated eaat3 upregulation in antioxidant defense and ferroptosis sensitivity in lung cancer
url https://doi.org/10.1038/s41419-025-07453-y
work_keys_str_mv AT donghuawen nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT wenjingli nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT xiangsong nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT minhu nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT yuelingliao nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT dongliangxu nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT jiongdeng nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer
AT wenzhengguo nfkbmediatedeaat3upregulationinantioxidantdefenseandferroptosissensitivityinlungcancer