Pharmacological inhibition of JNK-MAPK disrupts cigarette smoke-induced RUNX2/Galectin-3 -driven EMT and cancer stemness in lung adenocarcinoma cells
(2025) In Biochemical Pharmacology 242.- Abstract
Cigarette smoke (CS), a major driver of lung cancer (LC), promotes epithelial-mesenchymal transition (EMT) and stemness resulting in metastasis, therapy resistance, and recurrence, but the precise mechanism is elusive. Building on our earlier identification of Runt related transcription factor-2 (RUNX2) and Galectin-3 (Gal-3) as mediators of CS-induced EMT, in this study, we aimed to identify a potential molecular mechanism and delineate the upstream regulators of RUNX2 using A549 lung adenocarcinoma cells and human small airway epithelial cells (SAECs) cultured at the air–liquid interface (ALI). CSE exposure markedly elevated intracellular reactive oxygen species (ROS), assessed via Dichloro-dihydro-fluorescein diacetate (DCFH-DA)... (More)
Cigarette smoke (CS), a major driver of lung cancer (LC), promotes epithelial-mesenchymal transition (EMT) and stemness resulting in metastasis, therapy resistance, and recurrence, but the precise mechanism is elusive. Building on our earlier identification of Runt related transcription factor-2 (RUNX2) and Galectin-3 (Gal-3) as mediators of CS-induced EMT, in this study, we aimed to identify a potential molecular mechanism and delineate the upstream regulators of RUNX2 using A549 lung adenocarcinoma cells and human small airway epithelial cells (SAECs) cultured at the air–liquid interface (ALI). CSE exposure markedly elevated intracellular reactive oxygen species (ROS), assessed via Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay, and promoted invasive behavior (Boyden chamber assay), spheroid formation, and colony formation, the hallmarks of cancer stemness. Expression analysis via RT-qPCR, immunoblotting, and immunocytochemistry revealed that CSE upregulated EMT and stemness-associated markers, notably via upregulating RUNX2 and Galectin-3, at both transcriptional and translational levels through the involvement of c-Jun N-terminal kinase- Mitogen-Activated Protein Kinase (JNK-MAPK) pathways. A specific pharmacological inhibitor of JNK (SP600125) significantly attenuated CSE-induced RUNX2 and Galectin-3 (Gal-3) expression, and also reversed CSE-driven EMT marker alterations, suppressed transcriptional EMT perturbations, and reduced proinflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α). In conclusion, this study identifies that ROS/JNK/RUNX2/Gal-3 axis drives CS-induced oncogenic plasticity, suggesting that targeted inhibition of this pathway could be an effective strategy for mitigating CS-related LC progression.
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- author
- Chhetri, Karan ; Sharma, Jiten R. LU ; Vasita, Rajesh ; Singh, Rana P. and Yadav, Umesh C.S.
- organization
- publishing date
- 2025-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Cigarette smoke, Epithelial-mesenchymal transition (EMT), Galectin-3, JNK, Lung cancer, Reactive oxygen species (ROS), RUNX2, Stemness
- in
- Biochemical Pharmacology
- volume
- 242
- article number
- 117399
- publisher
- Elsevier
- external identifiers
-
- pmid:41047043
- scopus:105018077555
- ISSN
- 0006-2952
- DOI
- 10.1016/j.bcp.2025.117399
- language
- English
- LU publication?
- yes
- id
- 9ac6bb8d-d0b7-4037-b1cc-57806c37311e
- date added to LUP
- 2025-11-20 15:22:16
- date last changed
- 2025-11-20 15:22:50
@article{9ac6bb8d-d0b7-4037-b1cc-57806c37311e,
abstract = {{<p>Cigarette smoke (CS), a major driver of lung cancer (LC), promotes epithelial-mesenchymal transition (EMT) and stemness resulting in metastasis, therapy resistance, and recurrence, but the precise mechanism is elusive. Building on our earlier identification of Runt related transcription factor-2 (RUNX2) and Galectin-3 (Gal-3) as mediators of CS-induced EMT, in this study, we aimed to identify a potential molecular mechanism and delineate the upstream regulators of RUNX2 using A549 lung adenocarcinoma cells and human small airway epithelial cells (SAECs) cultured at the air–liquid interface (ALI). CSE exposure markedly elevated intracellular reactive oxygen species (ROS), assessed via Dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay, and promoted invasive behavior (Boyden chamber assay), spheroid formation, and colony formation, the hallmarks of cancer stemness. Expression analysis via RT-qPCR, immunoblotting, and immunocytochemistry revealed that CSE upregulated EMT and stemness-associated markers, notably via upregulating RUNX2 and Galectin-3, at both transcriptional and translational levels through the involvement of c-Jun N-terminal kinase- Mitogen-Activated Protein Kinase (JNK-MAPK) pathways. A specific pharmacological inhibitor of JNK (SP600125) significantly attenuated CSE-induced RUNX2 and Galectin-3 (Gal-3) expression, and also reversed CSE-driven EMT marker alterations, suppressed transcriptional EMT perturbations, and reduced proinflammatory cytokines, including monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α). In conclusion, this study identifies that ROS/JNK/RUNX2/Gal-3 axis drives CS-induced oncogenic plasticity, suggesting that targeted inhibition of this pathway could be an effective strategy for mitigating CS-related LC progression.</p>}},
author = {{Chhetri, Karan and Sharma, Jiten R. and Vasita, Rajesh and Singh, Rana P. and Yadav, Umesh C.S.}},
issn = {{0006-2952}},
keywords = {{Cigarette smoke; Epithelial-mesenchymal transition (EMT); Galectin-3; JNK; Lung cancer; Reactive oxygen species (ROS); RUNX2; Stemness}},
language = {{eng}},
publisher = {{Elsevier}},
series = {{Biochemical Pharmacology}},
title = {{Pharmacological inhibition of JNK-MAPK disrupts cigarette smoke-induced RUNX2/Galectin-3 -driven EMT and cancer stemness in lung adenocarcinoma cells}},
url = {{http://dx.doi.org/10.1016/j.bcp.2025.117399}},
doi = {{10.1016/j.bcp.2025.117399}},
volume = {{242}},
year = {{2025}},
}