{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T09:24:28Z","timestamp":1788341068902,"version":"build-2803163510"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T00:00:00Z","timestamp":1788307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["RS-2025-25431554"],"award-info":[{"award-number":["RS-2025-25431554"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"award":["RS-2025-25431554"],"award-info":[{"award-number":["RS-2025-25431554"]}],"id":[{"id":"https:\/\/ror.org\/013aysd81","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004230","name":"Korea Polar Research Institute","doi-asserted-by":"publisher","award":["PE26220"],"award-info":[{"award-number":["PE26220"]}],"id":[{"id":"10.13039\/501100004230","id-type":"DOI","asserted-by":"publisher"}]},{"award":["PE26220"],"award-info":[{"award-number":["PE26220"]}],"id":[{"id":"https:\/\/ror.org\/00n14a494","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Cancer immunotherapy faces significant challenges in treating \u201ccold\u201d tumors, which respond poorly to current strategies. Breast cancer is a major cause of cancer-related mortality in women and is a representative immunosuppressive cold tumor. The luminal A subtype of breast cancer exhibits an immune-cold phenotype, with MCF-7 cells used as a representative cell model. Here, we identified (3\u03b2,5\u03b1,8\u03b1)-5,8-epidioxyergost-6-en-3\u03b2-ol (EEO), isolated from Gymnopilus orientispectabilis, as an inducer of immunogenic cell death (ICD) hallmarks in MCF-7 cells, including cell surface exposure of calreticulin (CRT) and the extracellular release of damage-associated molecular patterns (DAMPs), such as ATP and High Mobility Group Box 1 (HMGB1). These ICD-associated events subsequently promoted CRT-dependent dendritic cell (DC) activation in a co-culture system with bone marrow-derived DCs and MCF-7 cells. Furthermore, surface-exposed CRT enhanced the phagocytosis of EEO-treated MCF-7 cells by DCs, and this phagocytic activity promoted DC maturation, as indicated by increased cell surface levels of the maturation markers major histocompatibility complex class II (MHC II) and cluster of differentiation 86 (CD86). Moreover, CRT knockdown in MCF-7 cells reduced surface CRT exposure following EEO treatment, thereby suppressing DC-mediated phagocytosis and subsequent DC maturation. Taken together, these findings suggest that EEO is a promising candidate for enhancing the antitumor efficacy of existing breast cancer immunotherapies through ICD-mediated DC maturation and converting immunologically cold tumors into hot tumors.<\/jats:p>","DOI":"10.3390\/life16091466","type":"journal-article","created":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T08:56:01Z","timestamp":1788339361000},"page":"1466","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Immunogenic Cell Death Induced by EEO in Breast Cancer Cells Promotes Dendritic Cell Activation in a CRT-Dependent Manner"],"prefix":"10.3390","volume":"16","author":[{"given":"Seong-Ah","family":"Shin","sequence":"first","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sun Young","family":"Moon","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seyeon","family":"Choi","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minji","family":"Kim","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moonsu","family":"Kim","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4831-2228","authenticated-orcid":false,"given":"Jun Hyuck","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Polar Sciences, University of Science and Technology, Incheon 21990, Republic of Korea"},{"name":"Research Unit of Cryogenic Novel Material, Korea Polar Research Institute, Incheon 21990, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Seulah","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Convergent Biotechnology and Advanced Materials Science, BK21 Interdisciplinary Program in IT-Bio Convergence System, Kyung Hee University, Yongin 17104, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5285-9138","authenticated-orcid":false,"given":"Ki Hyun","family":"Kim","sequence":"additional","affiliation":[{"name":"School of Pharmacy, Sungkyunkwan University, Suwon 440-746, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9928-0847","authenticated-orcid":false,"given":"Hyun Ho","family":"Park","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Chung Ang University, Seoul 06974, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9338-7934","authenticated-orcid":false,"given":"Ui Joung","family":"Youn","sequence":"additional","affiliation":[{"name":"Department of Polar Sciences, University of Science and Technology, Incheon 21990, Republic of Korea"},{"name":"Division of Life Sciences, Korea Polar Research Institute, Incheon 21990, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang Sup","family":"Lee","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, Jinju 52828, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e2400653","DOI":"10.1200\/GO-24-00653","article-title":"Assessing Breast Cancer Diagnosis and Management in Ghana per the Global Breast Cancer Initiative Key Performance Indicators","volume":"11","author":"Mensah","year":"2025","journal-title":"JCO Glob. Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1016\/S0140-6736(20)32381-3","article-title":"Breast cancer","volume":"397","author":"Loibl","year":"2021","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Dvir, K., Giordano, S., and Leone, J.P. (2024). Immunotherapy in Breast Cancer. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25147517"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Yang, L., Hu, Q., and Huang, T. (2024). Breast Cancer Treatment Strategies Targeting the Tumor Microenvironment: How to Convert \u201cCold\u201d Tumors to \u201cHot\u201d Tumors. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25137208"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1016\/j.trecan.2024.11.011","article-title":"Turning cold into hot: Emerging strategies to fire up the tumor microenvironment","volume":"11","author":"Ma","year":"2025","journal-title":"Trends Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"280","DOI":"10.7150\/ijbs.121356","article-title":"Mechanical Microenvironment in Tumor Immune Evasion: Bidirectional Regulation Between Matrix Stiffness and Immune Cells and Its Therapeutic Implications","volume":"22","author":"Ai","year":"2026","journal-title":"Int. J. Biol. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1038\/s41590-022-01132-2","article-title":"Immunogenic cell stress and death","volume":"23","author":"Kroemer","year":"2022","journal-title":"Nat. Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1038\/s41573-024-00920-9","article-title":"Targeting immunogenic cell stress and death for cancer therapy","volume":"23","author":"Galluzzi","year":"2024","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1146\/annurev-immunol-032712-100008","article-title":"Immunogenic cell death in cancer therapy","volume":"31","author":"Kroemer","year":"2013","journal-title":"Annu. Rev. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e27878","DOI":"10.4161\/onci.27878","article-title":"Trial Watch: Chemotherapy with immunogenic cell death inducers","volume":"3","author":"Vacchelli","year":"2014","journal-title":"Oncoimmunology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1186\/s13765-025-00986-y","article-title":"Ginsenoside Rh2 regulates cardiomyocyte autophagy-dependent apoptosis through the PI3K-Akt-mTOR signaling pathway to attenuate doxorubicin-induced cardiotoxicity","volume":"68","author":"Wu","year":"2025","journal-title":"Appl. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2437918","DOI":"10.1080\/21645515.2024.2437918","article-title":"Immunogenic cell death: A new strategy to enhancing cancer immunotherapy","volume":"20","author":"Dou","year":"2024","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Arimoto, K.I., Miyauchi, S., Liu, M., and Zhang, D.E. (2024). Emerging role of immunogenic cell death in cancer immunotherapy. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1390263"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1038\/nm1523","article-title":"Calreticulin exposure dictates the immunogenicity of cancer cell death","volume":"13","author":"Obeid","year":"2007","journal-title":"Nat. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/sj.cdd.4402269","article-title":"Molecular characteristics of immunogenic cancer cell death","volume":"15","author":"Tesniere","year":"2008","journal-title":"Cell Death Differ."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2994","DOI":"10.1002\/1878-0261.12851","article-title":"Targeting immunogenic cell death in cancer","volume":"14","author":"Ahmed","year":"2020","journal-title":"Mol. Oncol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1016\/j.apsb.2021.08.020","article-title":"Emerging role of natural products in cancer immunotherapy","volume":"12","author":"Dong","year":"2022","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"629","DOI":"10.1007\/s12272-019-01150-z","article-title":"Natural compound inducers of immunogenic cell death","volume":"42","author":"Diederich","year":"2019","journal-title":"Arch. Pharm. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"155828","DOI":"10.1016\/j.phymed.2024.155828","article-title":"Immunogenic cell death inducers for cancer therapy: An emerging focus on natural products","volume":"132","author":"Zhang","year":"2024","journal-title":"Phytomedicine"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Jung, B., Heo, E., Nguyen, D., Youn, U., Kim, K., Son, B., and Lee, S. (2025). Antimicrobial Steroids from Poisonous Mushroom Gymnopilus orientispectabilis and Their Molecular Docking Studies. Separations, 12.","DOI":"10.3390\/separations12020023"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1016\/j.imbio.2013.02.003","article-title":"Intravenous transfer of apoptotic cell-treated dendritic cells leads to immune tolerance by blocking Th17 cell activity","volume":"218","author":"Zhou","year":"2013","journal-title":"Immunobiology"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4854","DOI":"10.1111\/jcmm.14356","article-title":"Immunogenic cell death in cancer therapy: Present and emerging inducers","volume":"23","author":"Zhou","year":"2019","journal-title":"J. Cell Mol. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1038\/nri.2016.107","article-title":"Immunogenic cell death in cancer and infectious disease","volume":"17","author":"Galluzzi","year":"2017","journal-title":"Nat. Rev. Immunol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e1466765","DOI":"10.1080\/2162402X.2018.1466765","article-title":"PKR and GCN2 stress kinases promote an ER stress-independent eIF2alpha phosphorylation responsible for calreticulin exposure in melanoma cells","volume":"7","author":"Giglio","year":"2018","journal-title":"Oncoimmunology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"3399","DOI":"10.3892\/etm.2017.4989","article-title":"Calreticulin is an effective immunologic adjuvant to tumor-associated antigens","volume":"14","author":"Wang","year":"2017","journal-title":"Exp. Ther. Med."},{"key":"ref_26","first-page":"7288","article-title":"Dendritic cell activation and maturation induced by recombinant calreticulin fragment 39-272","volume":"8","author":"Li","year":"2015","journal-title":"Int. J. Clin. Exp. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1111\/j.1365-2249.2011.04423.x","article-title":"Modulatory role of calreticulin as chaperokine for dendritic cell-based immunotherapy","volume":"165","author":"Bajor","year":"2011","journal-title":"Clin. Exp. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.cell.2005.08.032","article-title":"Cell-surface calreticulin initiates clearance of viable or apoptotic cells through trans-activation of LRP on the phagocyte","volume":"123","author":"Gardai","year":"2005","journal-title":"Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/j.trecan.2020.02.022","article-title":"Turning Cold into Hot: Firing up the Tumor Microenvironment","volume":"6","author":"Duan","year":"2020","journal-title":"Trends Cancer"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"275","DOI":"10.5483\/BMBRep.2023-0024","article-title":"Immunogenic cell death in cancer immunotherapy","volume":"56","author":"Choi","year":"2023","journal-title":"BMB Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"753","DOI":"10.3892\/ijo.2016.3558","article-title":"Influence of doxorubicin on apoptosis and oxidative stress in breast cancer cell lines","volume":"49","author":"Calaf","year":"2016","journal-title":"Int. J. Oncol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1186\/s13765-025-01061-2","article-title":"ACF-01 enhances the anticancer effect of doxorubicin by suppressing stemness and inducing apoptosis in triple-negative breast cancer stem-like cells","volume":"68","author":"Yang","year":"2025","journal-title":"Appl. Biol. Chem."},{"key":"ref_33","first-page":"9687","article-title":"Mitoxantrone induces apoptosis in osteosarcoma cells through regulation of the Akt\/FOXO3 pathway","volume":"15","author":"Park","year":"2018","journal-title":"Oncol. Lett."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"14912","DOI":"10.1038\/s41598-025-98499-7","article-title":"Isoalantolactone induces the apoptosis of oxaliplatin-resistant human colorectal cancer cells mediated by ROS generation and activation of JNK and p38 MAPK","volume":"15","author":"Lee","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"41","DOI":"10.3892\/ijo_00000311","article-title":"gammaH2AX and cleaved PARP-1 as apoptotic markers in irradiated breast cancer BT474 cellular spheroids","volume":"35","author":"Qvarnstrom","year":"2009","journal-title":"Int. J. Oncol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Bezu, L., Gomes-de-Silva, L.C., Dewitte, H., Breckpot, K., Fucikova, J., Spisek, R., Galluzzi, L., Kepp, O., and Kroemer, G. (2015). Combinatorial strategies for the induction of immunogenic cell death. Front. Immunol., 6.","DOI":"10.3389\/fimmu.2015.00187"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Kielbik, M., Szulc-Kielbik, I., and Klink, M. (2021). Calreticulin-Multifunctional Chaperone in Immunogenic Cell Death: Potential Significance as a Prognostic Biomarker in Ovarian Cancer Patients. Cells, 10.","DOI":"10.3390\/cells10010130"},{"key":"ref_38","first-page":"47","article-title":"Immunogenic Cell Death: A Step Ahead of Autophagy in Cancer Therapy","volume":"3","author":"Gupta","year":"2021","journal-title":"J. Cancer Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1038\/nature08296","article-title":"Nucleotides released by apoptotic cells act as a find-me signal to promote phagocytic clearance","volume":"461","author":"Elliott","year":"2009","journal-title":"Nature"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1038\/s41419-020-03221-2","article-title":"Detection of immunogenic cell death and its relevance for cancer therapy","volume":"11","author":"Fucikova","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1573","DOI":"10.1007\/s00262-010-0833-1","article-title":"Toll-like receptor expression and function in human dendritic cell subsets: Implications for dendritic cell-based anti-cancer immunotherapy","volume":"59","author":"Schreibelt","year":"2010","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Hato, L., Vizcay, A., Eguren, I., Perez-Gracia, J.L., Rodriguez, J., Gallego Perez-Larraya, J., Sarobe, P., Inoges, S., Diaz de Cerio, A.L., and Santisteban, M. (2024). Dendritic Cells in Cancer Immunology and Immunotherapy. Cancers, 16.","DOI":"10.3390\/cancers16050981"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.cyto.2017.07.022","article-title":"IL-33-matured dendritic cells promote Th17 cell responses via IL-1beta and IL-6","volume":"99","author":"Park","year":"2017","journal-title":"Cytokine"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"639","DOI":"10.1016\/j.tcb.2015.08.003","article-title":"An Apoptotic \u2018Eat Me\u2019 Signal: Phosphatidylserine Exposure","volume":"25","author":"Segawa","year":"2015","journal-title":"Trends Cell Biol."}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/9\/1466\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T09:07:05Z","timestamp":1788340025000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/9\/1466"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,2]]},"references-count":44,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["life16091466"],"URL":"https:\/\/doi.org\/10.3390\/life16091466","relation":{},"ISSN":["2075-1729"],"issn-type":[{"value":"2075-1729","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,2]]}}}