{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T13:46:51Z","timestamp":1785764811777,"version":"3.56.0"},"reference-count":29,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T00:00:00Z","timestamp":1785542400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Gliomas are the most common group of primary brain tumors, among which glioblastoma multiforme (GBM) is characterized by a particularly poor prognosis and the limited effectiveness of available treatments. Ophiobolin A (OP-A), a natural sesterterpenoid, exhibits promising anticancer properties, including the ability to cross the blood-brain barrier and induce paraptosis-like cell death. However, the molecular mechanisms underlying its action, especially in the early phase of the cellular response, remain not fully understood. The aim of this study was to analyze early changes in the expression of genes associated with apoptosis, ferroptosis, and antioxidant mechanisms in T98G and U118MG glioma cells exposed to OP-A. Gene expression was assessed by RT-qPCR, apoptosis was evaluated using Annexin V\/PI staining and flow cytometry, and treatment-induced morphological changes were documented by brightfield microscopy. Statistical analysis was performed using the Mann\u2013Whitney U test. Descriptive Annexin V\/PI analysis showed a lower proportion of viable cells and a higher proportion of early apoptotic cells in the analyzed OP-A-treated T98G and U118MG samples compared with the corresponding vehicle-control samples. These preliminary observations were based on technical replicates from a single biological experiment and require confirmation in independent biological replicates. Transcriptional profiling revealed a shift toward a pro-apoptotic phenotype, characterized by increased BAX and FAS expression together with a trend toward reduced BCL2 expression, whereas ferroptosis-associated genes remained largely unchanged. Notably, SLC7A11 upregulation suggested activation of compensatory antioxidant mechanisms in response to OP-A treatment. In T98G cells, OP-A induced a distinct and reproducible transcriptional signature that enabled accurate discrimination from control conditions (AUC = 0.833). Feature importance and SHAP analyses identified BAX as the most informative predictor, followed by SLC7A11 and FAS, with bootstrap validation confirming BAX as a stable marker. Pathway analysis demonstrated selective activation of apoptosis- and cysteine metabolism-related pathways, while hierarchical clustering revealed that OP-A generated a transcriptional profile distinct from oxidative stress-inducing agents. The predictive performance of this molecular signature was cell-line dependent, showing weaker discrimination in U118MG cells.Short-term OP-A exposure in T98G and U118MG cells was associated with exploratory trends in apoptosis- and redox-related gene expression and a higher proportion of Annexin V-positive cells. The machine-learning analyses identified candidate discriminatory features within this limited dataset but should be regarded as hypothesis-generating. Larger studies with independent biological replication, additional GBM models, different exposure conditions, and functional validation are required to confirm these observations and clarify the mechanism of OP-A action.<\/jats:p>","DOI":"10.3390\/ijms27156891","type":"journal-article","created":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T12:57:56Z","timestamp":1785761876000},"page":"6891","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ophiobolin A Induces an Apoptotic Transcriptional Signature and Modulates Redox Homeostasis in T98G and U118MG Glioblastoma Cells: A Machine Learning Approach"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-6962-2655","authenticated-orcid":false,"given":"Pawe\u0142","family":"Wo\u017anicki","sequence":"first","affiliation":[{"name":"Doctoral School, Faculty of Medicine, Collegium Medicum, University of Rzesz\u00f3w, 35-310 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9110-2314","authenticated-orcid":false,"given":"Dorota","family":"Hudy","sequence":"additional","affiliation":[{"name":"Department of Medical and Molecular Biology, Faculty of Medical Science in Zabrze, Medical University of Silesia in Katowice, Jordana 19 Str., 41-808 Zabrze, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-8021-3766","authenticated-orcid":false,"given":"Oliwia","family":"Trzasko\u015b","sequence":"additional","affiliation":[{"name":"Department of Human Immunology, Faculty of Medicine, Collegium Medicum, University of Rzesz\u00f3w, 35-310 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Avijit","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8361-7318","authenticated-orcid":false,"given":"Marvin","family":"Xavierselvan","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1264-2882","authenticated-orcid":false,"given":"Jacek","family":"Tabarkiewicz","sequence":"additional","affiliation":[{"name":"Department of Human Immunology, Faculty of Medicine, Collegium Medicum, University of Rzesz\u00f3w, 35-310 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3686-5685","authenticated-orcid":false,"given":"Joanna Katarzyna","family":"Strzelczyk","sequence":"additional","affiliation":[{"name":"Department of Medical and Molecular Biology, Faculty of Medical Science in Zabrze, Medical University of Silesia in Katowice, Jordana 19 Str., 41-808 Zabrze, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2661-6570","authenticated-orcid":false,"given":"David","family":"Aebisher","sequence":"additional","affiliation":[{"name":"Department of Photomedicine and Physical Chemistry, Faculty of Medicine, Collegium Medicum, University of Rzeszow, 35-310 Rzesz\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"iv1","DOI":"10.1093\/neuonc\/noy131","article-title":"CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2011\u20132015","volume":"20","author":"Ostrom","year":"2018","journal-title":"Neuro-Oncology"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1007\/s00401-016-1545-1","article-title":"The 2016 World Health Organization Classification of Tumors of the Central Nervous System: A Summary","volume":"131","author":"Louis","year":"2016","journal-title":"Acta Neuropathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e32042","DOI":"10.1097\/MD.0000000000032042","article-title":"Clinical Characteristics and Prognosis of Patients with Glioblastoma: A Review of Survival Analysis of 1674 Patients Based on SEER Database","volume":"101","author":"Chen","year":"2022","journal-title":"Medicine"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1007\/s11060-019-03218-8","article-title":"Primary Central Nervous System Tumor Treatment and Survival in the United States, 2004\u20132015","volume":"144","author":"Garcia","year":"2019","journal-title":"J. Neurooncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1056\/NEJMoa043330","article-title":"Radiotherapy plus Concomitant and Adjuvant Temozolomide for Glioblastoma","volume":"352","author":"Stupp","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e315","DOI":"10.1016\/S1470-2045(17)30194-8","article-title":"European Association for Neuro-Oncology (EANO) Guideline on the Diagnosis and Treatment of Adult Astrocytic and Oligodendroglial Gliomas","volume":"18","author":"Weller","year":"2017","journal-title":"Lancet Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1186\/s12943-022-01513-z","article-title":"Glioma Targeted Therapy: Insight into Future of Molecular Approaches","volume":"21","author":"Yang","year":"2022","journal-title":"Mol. Cancer"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1021\/acs.jnatprod.9b01285","article-title":"Natural Products as Sources of New Drugs over the Nearly Four Decades from 01\/1981 to 09\/2019","volume":"83","author":"Newman","year":"2020","journal-title":"J. Nat. Prod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1016\/S0024-3205(00)00668-8","article-title":"The Biology of Ophiobolins","volume":"67","author":"Au","year":"2000","journal-title":"Life Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1039\/D3NP00035D","article-title":"The Incredible Story of Ophiobolin A and Sphaeropsidin A: Two Fungal Terpenes from Wilt-Inducing Phytotoxins to Promising Anticancer Compounds","volume":"41","author":"Evidente","year":"2024","journal-title":"Nat. Prod. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1080\/10717544.2019.1616235","article-title":"A Comprehensive Review in Improving Delivery of Small-Molecule Chemotherapeutic Agents Overcoming the Blood-Brain\/Brain Tumor Barriers for Glioblastoma Treatment","volume":"26","author":"Wang","year":"2019","journal-title":"Drug Deliv."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"106740","DOI":"10.18632\/oncotarget.22537","article-title":"Ophiobolin A Kills Human Glioblastoma Cells by Inducing Endoplasmic Reticulum Stress via Disruption of Thiol Proteostasis","volume":"8","author":"Kim","year":"2017","journal-title":"Oncotarget"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e561","DOI":"10.1038\/cddis.2013.85","article-title":"Ophiobolin A Induces Paraptosis-like Cell Death in Human Glioblastoma Cells by Decreasing BKCa Channel Activity","volume":"4","author":"Bury","year":"2013","journal-title":"Cell Death Dis."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1575","DOI":"10.1002\/cncr.23338","article-title":"Identification of Survival-Related Genes of the Phosphatidylinositol 3\u2019-Kinase Signaling Pathway in Glioblastoma Multiforme","volume":"112","author":"Ruano","year":"2008","journal-title":"Cancer"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"587","DOI":"10.3724\/abbs.2023068","article-title":"Ginsenoside Rh3 induces pyroptosis and ferroptosis through the Stat3\/p53\/NRF2 axis in colorectal cancer cells","volume":"55","author":"Wu","year":"2023","journal-title":"Acta Biochim. Biophys. Sin."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"G236","DOI":"10.1152\/ajpgi.00091.2010","article-title":"CHOP and AP-1 Cooperatively Mediate PUMA Expression during Lipoapoptosis","volume":"299","author":"Cazanave","year":"2010","journal-title":"Am. J. Physiol.-Gastrointest. Liver Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1128\/MCB.19.1.495","article-title":"CHOP-Dependent Stress-Inducible Expression of a Novel Form of Carbonic Anhydrase VI","volume":"19","author":"Sok","year":"1999","journal-title":"Mol. Cell. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11108","DOI":"10.1074\/jbc.M115.645333","article-title":"DDIT3 and KAT2A Proteins Regulate TNFRSF10A and TNFRSF10B Expression in Endoplasmic Reticulum Stress-Mediated Apoptosis in Human Lung Cancer Cells","volume":"290","author":"Li","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1111\/j.1750-3639.1999.tb00536.x","article-title":"Frequent Co-Alterations of TP53, P16\/CDKN2A, p14ARF, PTEN Tumor Suppressor Genes in Human Glioma Cell Lines","volume":"9","author":"Ishii","year":"1999","journal-title":"Brain Pathol."},{"key":"ref_20","first-page":"649","article-title":"Analysis of the P53 Gene and Its Expression in Human Glioblastoma Cells","volume":"54","author":"Kikuchi","year":"1994","journal-title":"Cancer Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1036","DOI":"10.1093\/jnci\/89.14.1036","article-title":"Characterization of P53 and P21 Functional Interactions in Glioma Cells En Route to Apoptosis","volume":"89","author":"Fueyo","year":"1997","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2256","DOI":"10.1016\/S0021-9258(17)35926-4","article-title":"Exchange of Cystine and Glutamate across Plasma Membrane of Human Fibroblasts","volume":"261","author":"Bannai","year":"1986","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.cell.2013.12.010","article-title":"Regulation of Ferroptotic Cancer Cell Death by GPX4","volume":"156","author":"Yang","year":"2014","journal-title":"Cell"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"424","DOI":"10.1038\/s41580-024-00703-5","article-title":"The Cell Biology of Ferroptosis","volume":"25","author":"Dixon","year":"2024","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_25","first-page":"883","article-title":"Increased Expression of Antioxidant Enzymes in Radioresistant Variant from U251 Human Glioblastoma Cell Line","volume":"13","author":"Lee","year":"2004","journal-title":"Int. J. Mol. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1111\/jnc.14250","article-title":"Glutathione Reductase Mediates Drug Resistance in Glioblastoma Cells by Regulating Redox Homeostasis","volume":"144","author":"Zhu","year":"2018","journal-title":"J. Neurochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"114350","DOI":"10.1016\/j.isci.2025.114350","article-title":"Systemic Cyst(e)Inase Administration Induces Ferroptosis and Synergizes with Temozolomide in Glioblastoma","volume":"29","author":"Ahmad","year":"2026","journal-title":"iScience"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5951","DOI":"10.1038\/onc.2015.60","article-title":"Drug Repurposing: Sulfasalazine Sensitizes Gliomas to Gamma Knife Radiosurgery by Blocking Cystine Uptake through System Xc\u2212, Leading to Glutathione Depletion","volume":"34","author":"Sleire","year":"2015","journal-title":"Oncogene"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1158\/1541-7786.MCR-16-0028","article-title":"Increased Expression of System Xc\u2212 in Glioblastoma Confers an Altered Metabolic State and Temozolomide Resistance","volume":"14","author":"Polewski","year":"2016","journal-title":"Mol. Cancer Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6891\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,3]],"date-time":"2026-08-03T13:00:54Z","timestamp":1785762054000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/15\/6891"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,1]]},"references-count":29,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["ijms27156891"],"URL":"https:\/\/doi.org\/10.3390\/ijms27156891","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,1]]}}}