Cancer-Associated Fibroblasts Hinder Lung Squamous Cell Carcinoma Oxidative Stress-Induced Apoptosis via METTL3 Mediated m6A Methylation of COL10A1.

{"sci_version": "1.2", "schema": "https://corpus.bntrg.co.uk/sci/1.0/sci.schema.json", "id": "pmid:36246404", "kind": "science-record", "revision": 5, "content_hash": "sha256:3596b075c94ee310740791acf713cba8201fe86334fe31e32e7babf0b5989e51", "created_at": "2026-11-01T18:15:37Z", "updated_at": "2026-11-01T22:50:22Z", "generator": {"name": "corpus-assay-engine", "run_id": "reading-sci-2026-11-01"}, "paper": {"state": "complete", "title": "Cancer-Associated Fibroblasts Hinder Lung Squamous Cell Carcinoma Oxidative Stress-Induced Apoptosis via METTL3 Mediated m6A Methylation of COL10A1.", "doi": "10.1155/2022/4320809", "pmid": "36246404", "authors": ["Yuchan Li", "Xiaoxue Li", "Muwen Deng", "Changda Ye", "Yuanhong Peng", "Yan Lu"], "journal": "Oxidative medicine and cellular longevity", "year": 2022, "abstract": "Background: Cancer-associated fibroblasts (CAFs) within the tumor microenvironment are key players in tumorigenesis and tumor development. Nevertheless, the regulatory mechanisms of CAFs on lung squamous cell carcinoma- (LUSC-) associated remain poorly elucidated.\nMethods: The microarray dataset GSE22874, containing 30 specimens of primary culture of normal fibroblasts (NFs) and 8 specimens of cancer-associated fibroblasts (CAFs) samples derived from LUSC, was retrieved from the Gene Expression Omnibus (GEO) database and then calculated by using the R language (limma package) to identify differentially expressed genes (DEGs). CAF-conditioned medium (CAF-CM) was collected and used to culture LUSC cells, followed by assessment of cell proliferation, apoptosis, and oxidative stress levels by using CCK-8, annexin V-FITC/PI double staining and ELISA assays. Subsequently, COL10A1 was knocked down in CAFs to assess the role of COL10A1 in CAF regulation of LUSC behavior. Bioinformatics online analysis and MeRIP were applied to predict and test the m6A modification of COL10A1 mRNA and the regulatory relationship with METTL3. Rescue experiments were next performed to explore the effects of METTL3 and COL10A1 in CAFs on LUSC cell proliferation, apoptosis, and oxidative stress. LUSC tumor cells with or without (COL10A1-silenced) CAFs were subcutaneously inoculated in nude mice to evaluate the effect of COL10A1 in CAFs on LUSC tumor growth.\nResults: Elevated expression of COL10A1 was found in LUSC-derived CAFs by GSE22874 dataset analysis. We discovered that COL10A1 and METTL3 was expressed in both LUSC cells and matched CAFs, while COL10A1 expression was prominently higher in CAFs than in LUSC cells. CAF-CM memorably encouraged LUSC cell proliferation and suppressed apoptosis-induced oxidative stress, which was reversed by interfering with COL10A1 expression in CAFs, suggesting that COL10A1 might be secreted by CAFs into the culture medium to exert its effects inside LUSC cells. Global m6A modification was decreased in METTL3 knocked down CAFs. M6A modification, expression levels, and stability of COL10A1 mRNA were impaired upon METTL3 knockdown in CAFs. Overexpression of COL10A1 in CAFs partially reversed the effect of METTL3 knockdown on the malignant behavior of LUSC cells. In vivo studies confirmed that CAFs accelerated LUSC tumor growth, and this effect was counteracted by COL10A1 silencing.\nConclusions: COL10A1 secreted by CAFs could facilitate LUSC cell proliferation and repress apoptosis-induced oxidative stress, and the mechanism was due to elevated expression mediated by METTL3 promoting its mRNA m6A modification, thereby accelerating tumor growth.", "claims": [{"id": "claim-method", "statement": "The difference analysis was performed using the limma package based on the R software, which employs the classical Bayesian's t-test analysis method with filtering criteria: |log fold change| ≥ 0.5, adjust P < 0.05.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36246404", "locator": "sections[6] \"2.4. Bioinformatics Analysis\" paragraph 2", "quote": "The difference analysis was performed using the limma package based on the R software, which employs the classical Bayesian's t-test analysis method with filtering criteria: |log fold change| ≥ 0.5, adjust P < 0.05."}], "state": "complete"}, {"id": "claim-data-access", "statement": "The labeled dataset used to support the findings of this study is available from the corresponding author upon request.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:36246404", "locator": "sections[27] \"Data Availability\" paragraph 2", "quote": "The labeled dataset used to support the findings of this study is available from the corresponding author upon request."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The GSE22874 analysis identified 20 differentially expressed genes between LUSC-derived CAFs and normal fibroblasts, with COL10A1 prominently highly expressed in CAFs.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:36246404", "locator": "sections[19] \"3.1. Elevated Expression of COL10A1 in CAFs of LUSC Based on GEO Dataset Mining\" paragraph 2", "quote": "In addition, GSE22874 dataset was found to have a total of 20 differentially expressed genes (DEGs) by screening (Figure 1(a))."}, {"asset": "source:pmid:36246404", "locator": "sections[19] \"3.1. Elevated Expression of COL10A1 in CAFs of LUSC Based on GEO Dataset Mining\" paragraph 2", "quote": "We found that COL10A1 was prominently highly expressed in CAFs (Figure 1(d))."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "complete", "objective": "The GSE22874 analysis identified 20 differentially expressed genes between LUSC-derived CAFs and normal fibroblasts, with COL10A1 prominently highly expressed in CAFs.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:36246404"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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LUSC tumor cells with", "raged LUSC cell proliferation and suppressed apoptosis-induced oxidative stress, which was reversed"], "order_basis": "paper_text_order:46", "id": "assay:1"}, {"role": "data_generation", "definition_id": "microarray", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["iated remain poorly elucidated. Methods: The microarray dataset GSE22874, containing 30 specimens of"], "order_basis": "paper_text_order:382", "id": "assay:2"}, {"role": "data_generation", "definition_id": "elisa", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.93, "evidence": ["CCK-8, annexin V-FITC/PI double staining and ELISA assays. Subsequently, COL10A1 was knocked do"], "order_basis": "paper_text_order:955", "id": "assay:3"}, {"role": "data_generation", "definition_id": "transfection", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.99, "evidence": ["ty of COL10A1 mRNA were impaired upon METTL3 knockdown in CAFs. Overexpression of COL10A1 in CAFs p", "CAFs partially reversed the effect of METTL3 knockdown on the malignant behavior of LUSC cells. In"], "order_basis": "paper_text_order:2266", "id": "assay:4"}, {"role": "data_generation", "definition_id": "animal_model", "definition_version": "99e1195c7d6f", "definition_digest": null, "required_operations": [], "support_state": "partial", "source_hashes": {"assay_lexicon": "99e1195c7d6f"}, "confidence": 0.77, "evidence": ["own on the malignant behavior of LUSC cells. 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