1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner

ABSTRACT The carcinogenic potential of the environmental pollutant 1,4‐dioxane (1,4‐D) in humans is not yet fully understood or recognised. In this study, we provide evidence that 1,4‐D acts as a carcinogen in human epithelial cells. Using the human bronchial epithelial cell line BEAS‐2B, with or wi...

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Main Authors: Ziwei Wang, Chitra Thakur, Zhuoyue Bi, Yiran Qiu, Wenxuan Zhang, Haoyan Ji, Arjun K. Venkatesan, Sashank Cherukuri, Ke Jian Liu, John D. Haley, Xinwei Mao, Jaymie Meliker, Fei Chen
Format: Article
Language:English
Published: Wiley 2025-05-01
Series:Journal of Extracellular Vesicles
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Online Access:https://doi.org/10.1002/jev2.70072
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author Ziwei Wang
Chitra Thakur
Zhuoyue Bi
Yiran Qiu
Wenxuan Zhang
Haoyan Ji
Arjun K. Venkatesan
Sashank Cherukuri
Ke Jian Liu
John D. Haley
Xinwei Mao
Jaymie Meliker
Fei Chen
author_facet Ziwei Wang
Chitra Thakur
Zhuoyue Bi
Yiran Qiu
Wenxuan Zhang
Haoyan Ji
Arjun K. Venkatesan
Sashank Cherukuri
Ke Jian Liu
John D. Haley
Xinwei Mao
Jaymie Meliker
Fei Chen
author_sort Ziwei Wang
collection DOAJ
description ABSTRACT The carcinogenic potential of the environmental pollutant 1,4‐dioxane (1,4‐D) in humans is not yet fully understood or recognised. In this study, we provide evidence that 1,4‐D acts as a carcinogen in human epithelial cells. Using the human bronchial epithelial cell line BEAS‐2B, with or without CRISPR‐Cas9‐mediated Nrf2 knockout, we demonstrate that continuous exposure to environmentally relevant concentrations of 1.25–20 ppm 1,4‐D over 2 months induces malignant transformation in an Nrf2‐dependent manner. Transformed cells exhibit enhanced anchorage‐independent growth in soft agar, increased migration and invasion, and tumorigenic potential in a xenograft mouse model. Integrated RNA sequencing and proteomics analyses reveal that 1,4‐D robustly activates Nrf2 signalling, driving extracellular vesicle (EV) biogenesis and cargo loading with syndecan 4 (SDC4) and other proteins, including COL12A1, CAPG and NNMT, which are associated with epithelial‐mesenchymal transition (EMT) and cancer metastasis. Nrf2 knockout reduces SDC4 expression and its incorporation into EVs, leading to decreased EV uptake by recipient cells. Unlike EVs from 1,4‐D‐transformed WT cells, which enhance the proliferation, migration and invasion of recipient cells, EVs from 1,4‐D‐transformed Nrf2 KO cells exhibit a diminished capacity to promote these EMT properties. Furthermore, we demonstrate that the Nrf2 target gene SDC4, induced by 1,4‐D and enriched in EVs, plays a critical role in EV uptake by recipient cells, thereby facilitating EMT propagation. Collectively, our findings suggest that 1,4‐D is a human carcinogen, with its carcinogenicity largely dependent on Nrf2 activation, which orchestrates the biogenesis of EVs with EMT‐promoting functions.
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spelling doaj-art-ab89ec3ff5b44e459183852906d9f5862025-08-20T03:48:22ZengWileyJournal of Extracellular Vesicles2001-30782025-05-01145n/an/a10.1002/jev2.700721,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent MannerZiwei Wang0Chitra Thakur1Zhuoyue Bi2Yiran Qiu3Wenxuan Zhang4Haoyan Ji5Arjun K. Venkatesan6Sashank Cherukuri7Ke Jian Liu8John D. Haley9Xinwei Mao10Jaymie Meliker11Fei Chen12Stony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USACivil Engineering, School of Marine and Atmospheric Sciences Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USADepartment of Civil Engineering, College of Engineering and Applied Sciences Stony Brook University Stony Brook New York USADepartment of Family, Population and Preventive Medicine, Renaissance School of Medicine Stony Brook University Stony Brook New York USAStony Brook Cancer Center, Department of Pathology, Renaissance School of Medicine Stony Brook University Stony Brook New York USAABSTRACT The carcinogenic potential of the environmental pollutant 1,4‐dioxane (1,4‐D) in humans is not yet fully understood or recognised. In this study, we provide evidence that 1,4‐D acts as a carcinogen in human epithelial cells. Using the human bronchial epithelial cell line BEAS‐2B, with or without CRISPR‐Cas9‐mediated Nrf2 knockout, we demonstrate that continuous exposure to environmentally relevant concentrations of 1.25–20 ppm 1,4‐D over 2 months induces malignant transformation in an Nrf2‐dependent manner. Transformed cells exhibit enhanced anchorage‐independent growth in soft agar, increased migration and invasion, and tumorigenic potential in a xenograft mouse model. Integrated RNA sequencing and proteomics analyses reveal that 1,4‐D robustly activates Nrf2 signalling, driving extracellular vesicle (EV) biogenesis and cargo loading with syndecan 4 (SDC4) and other proteins, including COL12A1, CAPG and NNMT, which are associated with epithelial‐mesenchymal transition (EMT) and cancer metastasis. Nrf2 knockout reduces SDC4 expression and its incorporation into EVs, leading to decreased EV uptake by recipient cells. Unlike EVs from 1,4‐D‐transformed WT cells, which enhance the proliferation, migration and invasion of recipient cells, EVs from 1,4‐D‐transformed Nrf2 KO cells exhibit a diminished capacity to promote these EMT properties. Furthermore, we demonstrate that the Nrf2 target gene SDC4, induced by 1,4‐D and enriched in EVs, plays a critical role in EV uptake by recipient cells, thereby facilitating EMT propagation. Collectively, our findings suggest that 1,4‐D is a human carcinogen, with its carcinogenicity largely dependent on Nrf2 activation, which orchestrates the biogenesis of EVs with EMT‐promoting functions.https://doi.org/10.1002/jev2.700721,4‐dioxaneEMTEV biogenesisNrf2SDC4
spellingShingle Ziwei Wang
Chitra Thakur
Zhuoyue Bi
Yiran Qiu
Wenxuan Zhang
Haoyan Ji
Arjun K. Venkatesan
Sashank Cherukuri
Ke Jian Liu
John D. Haley
Xinwei Mao
Jaymie Meliker
Fei Chen
1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
Journal of Extracellular Vesicles
1,4‐dioxane
EMT
EV biogenesis
Nrf2
SDC4
title 1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
title_full 1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
title_fullStr 1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
title_full_unstemmed 1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
title_short 1,4‐Dioxane Induces Epithelial‐Mesenchymal Transition and Carcinogenesis in an Nrf2‐Dependent Manner
title_sort 1 4 dioxane induces epithelial mesenchymal transition and carcinogenesis in an nrf2 dependent manner
topic 1,4‐dioxane
EMT
EV biogenesis
Nrf2
SDC4
url https://doi.org/10.1002/jev2.70072
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