{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T05:52:30Z","timestamp":1782885150923,"version":"3.54.5"},"reference-count":43,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,6,27]],"date-time":"2026-06-27T00:00:00Z","timestamp":1782518400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Scientific Research Deanship at University of Ha\u2019il\u2014Saudi Arabia","award":["RG-25 012"],"award-info":[{"award-number":["RG-25 012"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Objective: Lung cancer (LC) remains a leading cause of cancer mortality worldwide. Thymoquinone (TQ), a bioactive compound derived from Nigella sativa, possesses anti-inflammatory and antioxidant properties, but its precise mechanisms concerning miRNA regulation in LC are poorly defined. This study investigates the anti-cancer potential of TQ through modulation of microRNA signaling in LC. Methods: We employed an integrated approach combining bioinformatic predictions with rigorous experimental validation in A549 lung adenocarcinoma cells and SHP-77 human small-cell lung carcinoma (SCLC) cells. Bioinformatic analyses predicted miRNA targets, and experimental techniques included dual-luciferase reporter assays, miRNA inhibition, TaqMan RT-qPCR, cell-based ELISA, and Western blotting to dissect the molecular pathway. Results: We identified the transcription factor BACH1 as a direct and novel target of hsa-miR-155-5p. TQ potently suppressed interferon-\u03b3-induced expression of both hsa-miR-155-5p and its target, BACH1. This TQ-mediated suppression led to subsequent downregulation of the key metastasis-promoter Matrix Metalloproteinase-9 (MMP-9). Genetic inhibition of miR-155-5p or direct BACH1 inhibition phenocopied the effects of TQ, confirming the functional significance of this axis. Thus, we define a novel oncogenic signaling cascade\u2014the hsa-miR-155-5p\/BACH1\/MMP-9 axis that is effectively disrupted by TQ. Conclusions: This represents the first evidence that TQ exerts its anti-cancer effects in LC through the modulation of the critical signaling cascade (hsa-miR-155-5p \u2192 BACH1 \u2192 MMP-9). Our findings establish TQ as a multi-targeted agent capable of simultaneously inhibiting miRNA-mediated oncogenic signaling and protein-level effectors. The dual therapeutic action of TQ represents a novel therapeutic strategy and underscores its potential for synergistic combination therapies.<\/jats:p>","DOI":"10.3390\/biom16070955","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T03:06:34Z","timestamp":1782875194000},"page":"955","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeting the hsa-miR-155-5p\u2013BACH1\u2013MMP-9 Signaling Hub in Lung Cancer: A Novel Anticancer Mechanism of Thymoquinone"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8765-1428","authenticated-orcid":false,"given":"Yusuf Saleem","family":"Khan","sequence":"first","affiliation":[{"name":"Department of Anatomy, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4631-2769","authenticated-orcid":false,"given":"Aisha","family":"Farhana","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alfatih Mohamed Ahmed","family":"Alnajib","sequence":"additional","affiliation":[{"name":"Department of Surgery, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1594-1826","authenticated-orcid":false,"given":"Azharuddin Sajid Syed","family":"Khaja","sequence":"additional","affiliation":[{"name":"Department of Pathology, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hatim Adam","family":"Nagi","sequence":"additional","affiliation":[{"name":"Department of Family and Community Medicine, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tarig","family":"Ginawi","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abuzar Abdulwahab","family":"Osman","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, College of Medicine, University of Ha\u2019il, Ha\u2019il 55476, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8467-9692","authenticated-orcid":false,"given":"Ayman Ali Mohammed","family":"Alameen","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emad","family":"Manni","sequence":"additional","affiliation":[{"name":"Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Jouf University, Sakaka 72388, Saudi Arabia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8651-4218","authenticated-orcid":false,"given":"Zafar","family":"Rasheed","sequence":"additional","affiliation":[{"name":"LTT-LabTech Training Centre, 28 Russell St, South Brisbane, QLD 4101, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"535","DOI":"10.1016\/S0140-6736(21)00312-3","article-title":"Lung cancer","volume":"398","author":"Thai","year":"2021","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1007\/s10555-024-10187-6","article-title":"Healthcare disparities, screening, and molecular testing in the changing landscape of non-small cell lung cancer in the United States: A review","volume":"43","author":"Kurzrock","year":"2024","journal-title":"Cancer Metastasis Rev."},{"key":"ref_3","first-page":"45","article-title":"Epidemiology of lung cancer","volume":"25","author":"Thandra","year":"2021","journal-title":"Contemp. 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