{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T05:09:57Z","timestamp":1789016997860,"version":"build-2803163510"},"reference-count":48,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T00:00:00Z","timestamp":1788480000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004704","name":"National Research Council of Thailand","doi-asserted-by":"publisher","award":["NRCT5-RGJ63004-083."],"award-info":[{"award-number":["NRCT5-RGJ63004-083."]}],"id":[{"id":"10.13039\/501100004704","id-type":"DOI","asserted-by":"publisher"}]},{"award":["NRCT5-RGJ63004-083."],"award-info":[{"award-number":["NRCT5-RGJ63004-083."]}],"id":[{"id":"https:\/\/ror.org\/018wfhg78","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002842","name":"Chiang Mai University","doi-asserted-by":"publisher","award":["031\/2564"],"award-info":[{"award-number":["031\/2564"]}],"id":[{"id":"10.13039\/501100002842","id-type":"DOI","asserted-by":"publisher"}]},{"award":["031\/2564"],"award-info":[{"award-number":["031\/2564"]}],"id":[{"id":"https:\/\/ror.org\/05m2fqn25","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antioxidants"],"abstract":"<jats:p>High-grade osteosarcoma presents a significant clinical challenge due to unpredictable therapeutic responses and aggressive progression. This study aimed to identify the critical molecular pathways driving chemotherapy resistance and aggressive phenotypes in osteosarcoma patients. Through transcriptomic and bioinformatic analyses, we identified cholesterol biosynthesis as a key upregulated metabolic pathway in poor chemotherapy responders, where squalene epoxidase (SQLE) emerged as an exploratory candidate hub gene whose elevated expression significantly correlates with shortened survival in the TCGA cohort. We validated these findings by administering terbinafine, a known SQLE inhibitor. In highly chemoresistant SaOS-2 cells exhibiting the highest baseline SQLE expression, terbinafine synergistically sensitized cells to doxorubicin by driving cell death partly through apoptosis, as confirmed by caspase inhibition. The combination also promoted ferroptosis, indicated by elevated ROS and MDA along with decreased FSP1 and GPX4 expression. Furthermore, the co-treatment effectively suppressed clonogenic potential, induced G2\/M phase cell cycle arrest, and inhibited metastatic progression. These effects were mediated by the modulation of cell proliferation, metastasis, and survival genes through the coordinated regulation of the PI3K\/AKT\/mTOR, ERK, and JNK signaling cascades. Together, these results highlight the therapeutic potential of targeting the SQLE pathway to overcome doxorubicin resistance and suppress aggressive progression in high-grade osteosarcoma.<\/jats:p>","DOI":"10.3390\/antiox15091122","type":"journal-article","created":{"date-parts":[[2026,9,4]],"date-time":"2026-09-04T16:07:32Z","timestamp":1788538052000},"page":"1122","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Targeting SQLE-Mediated Cholesterol Metabolism to Promote Oxidative Stress and Attenuate Drug Resistance in Osteosarcoma"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-4782-6985","authenticated-orcid":false,"given":"Amonnat","family":"Sukhamwang","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dumnoensun","family":"Pruksakorn","sequence":"additional","affiliation":[{"name":"Center of Multidisciplinary Technology for Advanced Medicine (CMUTEAM), Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"},{"name":"Musculoskeletal Science and Translational Research (MSTR) Center, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"},{"name":"Department of Orthopedics, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8732-8911","authenticated-orcid":false,"given":"Pornngarm","family":"Dejkriengkraikul","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"},{"name":"Anticarcinogenesis and Apoptosis Research Cluster, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1693-1761","authenticated-orcid":false,"given":"Michael A.","family":"Dengler","sequence":"additional","affiliation":[{"name":"Division of Oncology, Department of Internal Medicine, Medical University of Graz, Auenbruggerplatz 15, 8036 Graz, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9023-2283","authenticated-orcid":false,"given":"Supachai","family":"Yodkeeree","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"},{"name":"Anticarcinogenesis and Apoptosis Research Cluster, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s12938-021-00860-0","article-title":"Osteosarcoma: A review of current and future therapeutic approaches","volume":"20","author":"Zhao","year":"2021","journal-title":"Biomed. Eng. OnLine"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1186\/s12957-025-04115-3","article-title":"Comparative outcomes of neoadjuvant chemotherapy versus upfront surgical resection in osteosarcoma: A systematic review and meta-analysis","volume":"23","author":"Alaseem","year":"2025","journal-title":"World J. Surg. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1007\/s12672-025-02208-9","article-title":"Systemic strategies for osteosarcoma: Advances and future directions","volume":"16","author":"Morya","year":"2025","journal-title":"Discov. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e15582","DOI":"10.1097\/MD.0000000000015582","article-title":"Methotrexate, doxorubicin, and cisplatinum regimen is still the preferred option for osteosarcoma chemotherapy: A meta-analysis and clinical observation","volume":"98","author":"Yu","year":"2019","journal-title":"Medicine"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Zhra, M., Akhund, S.A., and Mohammad, K.S. (2025). Advancements in Osteosarcoma Therapy: Overcoming Chemotherapy Resistance and Exploring Novel Pharmacological Strategies. Pharmaceuticals, 18.","DOI":"10.3390\/ph18040520"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"119","DOI":"10.3892\/ijo.2023.5567","article-title":"Mechanism of multidrug resistance to chemotherapy mediated by P-glycoprotein (Review)","volume":"63","author":"Tian","year":"2023","journal-title":"Int. J. Oncol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Belisario, D.C., Akman, M., Godel, M., Campani, V., Patrizio, M.P., Scotti, L., Hattinger, C.M., De Rosa, G., Donadelli, M., and Serra, M. (2020). ABCA1\/ABCB1 Ratio Determines Chemo- and Immune-Sensitivity in Human Osteosarcoma. Cells, 9.","DOI":"10.3390\/cells9030647"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"10535","DOI":"10.1200\/jco.2014.32.15_suppl.10535","article-title":"Impact of excision repair cross-complementation group 1 (ERCC1) protein on survival of patients with osteosarcoma treated with cisplatin-based chemotherapy","volume":"32","author":"Igarashi","year":"2014","journal-title":"J. Clin. Oncol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e18","DOI":"10.1002\/pdi3.18","article-title":"Metabolic reprogramming in osteosarcoma","volume":"1","author":"Shi","year":"2023","journal-title":"Pediatr. Discov."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1126","DOI":"10.1093\/annonc\/mdg286","article-title":"Neoadjuvant chemotherapy for osteosarcoma of the extremities with metastases at presentation: Recent experience at the Rizzoli Institute in 57 patients treated with cisplatin, doxorubicin, and a high dose of methotrexate and ifosfamide","volume":"14","author":"Bacci","year":"2003","journal-title":"Ann. Oncol."},{"key":"ref_11","first-page":"1","article-title":"Outcomes in treatment-na\u00efve patients with metastatic extremity osteosarcoma treated with OGS-12, a novel non\u2013high-dose methotrexate\u2013based, dose-dense combination chemotherapy, in a tertiary care cancer center","volume":"4","author":"Bajpai","year":"2018","journal-title":"J. Glob. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Sukhamwang, A., Pruksakorn, D., Dejkriengkraikul, P., Sangphukieo, A., Dissook, S., and Yodkeeree, S. (2025). The Cholesterol Biosynthesis Pathway Plays an Important Role in Chemotherapeutic Drug Response and Metastasis in High-Grade Osteosarcoma. Cells, 14.","DOI":"10.3390\/cells14130993"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.cpt.2025.07.001","article-title":"Chemoresistance: The hidden barrier in cancer treatment","volume":"4","author":"Dhiman","year":"2026","journal-title":"Cancer Pathog. Ther."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"EMBR202152537","DOI":"10.15252\/embr.202152537","article-title":"p53 transcriptionally regulates SQLE to repress cholesterol synthesis and tumor growth","volume":"22","author":"Sun","year":"2021","journal-title":"EMBO Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"19435","DOI":"10.1038\/srep19435","article-title":"Squalene epoxidase is a bona fide oncogene by amplification with clinical relevance in breast cancer","volume":"6","author":"Brown","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5066","DOI":"10.1038\/s41467-021-25325-9","article-title":"MiR-205-driven downregulation of cholesterol biosynthesis through SQLE-inhibition identifies therapeutic vulnerability in aggressive prostate cancer","volume":"12","author":"Kalogirou","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"497","DOI":"10.1038\/s41419-023-05987-7","article-title":"SQLE promotes pancreatic cancer growth by attenuating ER stress and activating lipid rafts-regulated Src\/PI3K\/Akt signaling pathway","volume":"14","author":"Xu","year":"2023","journal-title":"Cell Death Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"22646","DOI":"10.1038\/s41598-024-72506-9","article-title":"Squalene monooxygenase (SQLE) protects ovarian cancer cells from ferroptosis","volume":"14","author":"Zhang","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4796","DOI":"10.1038\/s41388-025-03626-3","article-title":"SQLE drives bladder cancer progression by boosting mitochondrial oxidative phosphorylation","volume":"44","author":"Dong","year":"2025","journal-title":"Oncogene"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"217586","DOI":"10.1016\/j.canlet.2025.217586","article-title":"SQLE-catalyzed squalene metabolism promotes mitochondrial biogenesis and tumor development in K-ras-driven cancer","volume":"616","author":"Pan","year":"2025","journal-title":"Cancer Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"101993","DOI":"10.1016\/j.mcp.2024.101993","article-title":"SQLE promotes osteosarcoma progression via activating TGF\u03b2\/SMAD signaling pathway","volume":"78","author":"Song","year":"2024","journal-title":"Mol. Cell. Probes"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.ejphar.2014.07.025","article-title":"Cisplatin in cancer therapy: Molecular mechanisms of action","volume":"740","author":"Dasari","year":"2014","journal-title":"Eur. J. Pharmacol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Kciuk, M., Gieleci\u0144ska, A., Mujwar, S., Ko\u0142at, D., Ka\u0142uzi\u0144ska-Ko\u0142at, \u017b., Celik, I., and Kontek, R. (2023). Doxorubicin-An Agent with Multiple Mechanisms of Anticancer Activity. Cells, 12.","DOI":"10.3390\/cells12040659"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Radeva, L., and Yoncheva, K. (2025). Doxorubicin Toxicity and Recent Approaches to Alleviating Its Adverse Effects with Focus on Oxidative Stress. Molecules, 30.","DOI":"10.3390\/molecules30153311"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2414923","DOI":"10.1002\/advs.202414923","article-title":"Disturbing Cholesterol\/Sphingolipid Metabolism by Squalene Epoxidase Arises Crizotinib Hepatotoxicity","volume":"12","author":"Yan","year":"2025","journal-title":"Adv. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1038\/s44303-025-00110-1","article-title":"Phosphatidylserine exposure and plasma membrane perforation as ferroptotic signatures for in vivo imaging","volume":"3","author":"Sonay","year":"2025","journal-title":"npj Imaging"},{"key":"ref_27","unstructured":"(2013). Medical use of squalene as a natural antioxidant. J. Marmara Univ. Inst. Health Sci., 3, 221\u2013229."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Bojko, A., Staniak, K., Czarnecka-Herok, J., Sunderland, P., Dudkowska, M., \u015aliwi\u0144ska, M.A., Salmina, K., and Sikora, E. (2020). Improved Autophagic Flux in Escapers from Doxorubicin-Induced Senescence\/Polyploidy of Breast Cancer Cells. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21176084"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"882","DOI":"10.1016\/j.neo.2015.11.008","article-title":"Polyploidy Formation in Doxorubicin-Treated Cancer Cells Can Favor Escape from Senescence","volume":"17","author":"Mosieniak","year":"2015","journal-title":"Neoplasia"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1186\/s12935-025-04079-0","article-title":"Etiology of polyploid giant cancer cells: A new frontier in cancer biology","volume":"26","author":"Manickasamy","year":"2026","journal-title":"Cancer Cell Int."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e2522063123","DOI":"10.1073\/pnas.2522063123","article-title":"Growth under pressure: The pros and cons of polyploidy induced by stress","volume":"123","author":"Oliver","year":"2026","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1038\/s41420-024-02206-w","article-title":"Polyploidy mitigates the impact of DNA damage while simultaneously bearing its burden","volume":"10","author":"Hayashi","year":"2024","journal-title":"Cell Death Discov."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Zhao, Y.-C., Li, Y.-F., Qiu, L., Jin, S.-Z., Shen, Y.-N., Zhang, C.-H., Cui, J., and Wang, T.-J. (2024). SQLE\u2014A promising prognostic biomarker in cervical cancer: Implications for tumor malignant behavior, cholesterol synthesis, epithelial-mesenchymal transition, and immune infiltration. BMC Cancer, 24.","DOI":"10.1186\/s12885-024-12897-0"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Zhang, L., Cao, Z., Hong, Y., He, H., Chen, L., Yu, Z., and Gao, Y. (2024). Squalene Epoxidase: Its Regulations and Links with Cancers. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25073874"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Kim, J.H., Kim, C.N., and Kang, D.W. (2019). Squalene Epoxidase Correlates E-Cadherin Expression and Overall Survival in Colorectal Cancer Patients: The Impact on Prognosis and Correlation to Clinicopathologic Features. J. Clin. Med., 8.","DOI":"10.3390\/jcm8050632"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1186\/s12943-023-01827-6","article-title":"PI3K\/AKT\/mTOR signaling transduction pathway and targeted therapies in cancer","volume":"22","author":"Glaviano","year":"2023","journal-title":"Mol. Cancer"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Yang, C., Tian, Y., Zhao, F., Chen, Z., Su, P., Li, Y., and Qian, A. (2020). Bone Microenvironment and Osteosarcoma Metastasis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21196985"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Xiang, Y., Yang, Y., Liu, J., and Yang, X. (2023). Functional role of MicroRNA\/PI3K\/AKT axis in osteosarcoma. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1219211"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Tam, S.Y., and Law, H.K. (2021). JNK in Tumor Microenvironment: Present Findings and Challenges in Clinical Translation. Cancers, 13.","DOI":"10.3390\/cancers13092196"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"465","DOI":"10.3109\/10799893.2015.1122045","article-title":"JNK pathway in osteosarcoma: Pathogenesis and therapeutics","volume":"36","author":"Li","year":"2016","journal-title":"J. Recept. Signal Transduct."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Varga, D., Hajdin\u00e1k, P., Makk-Merczel, K., and Szarka, A. (2022). The Possible Connection of Two Dual Function Processes: The Relationship of Ferroptosis and the JNK Pathway. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231911004"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1002\/ijc.11194","article-title":"In vitro and in vivo studies of the anticancer action of terbinafine in human cancer cell lines: G0\/G1 p53-associated cell cycle arrest","volume":"106","author":"Lee","year":"2003","journal-title":"Int. J. Cancer"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3284","DOI":"10.1016\/j.ymthe.2022.06.015","article-title":"Terbinafine prevents colorectal cancer growth by inducing dNTP starvation and reducing immune suppression","volume":"30","author":"Hu","year":"2022","journal-title":"Mol. Ther."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1002\/mc.20800","article-title":"Terbinafine inhibits oral squamous cell carcinoma growth through anti-cancer cell proliferation and anti-angiogenesis","volume":"51","author":"Chien","year":"2012","journal-title":"Mol. Carcinog."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"99","DOI":"10.3390\/jmp4020011","article-title":"Different Subtypes of Osteosarcoma: Histopathological Patterns and Clinical Behaviour","volume":"4","author":"Rothzerg","year":"2023","journal-title":"J. Mol. Pathol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1177\/15353702221080131","article-title":"Single-cell RNA-seq identification of four differentially expressed survival-related genes by a TARGET: Osteosarcoma database analysis","volume":"247","author":"Feleke","year":"2022","journal-title":"Exp. Biol. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"979","DOI":"10.3727\/096504018X15336368805108","article-title":"Exosomal miR-1228 from cancer-associated fibroblasts promotes cell migration and invasion of osteosarcoma by directly targeting SCAI","volume":"27","author":"Wang","year":"2019","journal-title":"Oncol. Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"100660","DOI":"10.1016\/j.jbo.2025.100660","article-title":"Lipid metabolic reprogramming and associated ferroptosis in osteosarcoma: From molecular mechanisms to potential targets","volume":"51","author":"Yin","year":"2025","journal-title":"J. Bone Oncol."}],"container-title":["Antioxidants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3921\/15\/9\/1122\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T04:41:38Z","timestamp":1789015298000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3921\/15\/9\/1122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,4]]},"references-count":48,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["antiox15091122"],"URL":"https:\/\/doi.org\/10.3390\/antiox15091122","relation":{},"ISSN":["2076-3921"],"issn-type":[{"value":"2076-3921","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,4]]}}}