{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T08:31:45Z","timestamp":1787646705028,"version":"build-2736575974"},"reference-count":32,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T00:00:00Z","timestamp":1787616000000},"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>Anti-CD226 antibody-mediated stimulation of NK-92 cells (sNK-92) represents a potential immunotherapeutic approach; however, its cytotoxic and apoptosis-associated effects in hormone-independent solid tumors remain insufficiently characterized. This study investigated the activity of sNK-92 cells against PC3 castration-resistant prostate cancer and SH-SY5Y neuroblastoma cell lines, using PNT1A normal prostate epithelial and BJ normal dermal fibroblast cells as non-malignant controls. Cytotoxicity was assessed by CCK-8 assay at target-to-effector (T:E) ratios of 1:1, 1:5, and 1:10, and markers historically associated with the intrinsic (BAX, caspase-9), extrinsic (caspase-8), and executioner (caspase-3) apoptotic pathways were evaluated quantitatively by ImageJ-based corrected total cell fluorescence (CTCF) analysis. sNK-92 cells produced significant ratio-dependent cytotoxicity against PC3 and SH-SY5Y cells, reaching 23.76% and 26.29%, respectively, at the 1:10 T:E ratio, with sNK-92 producing significantly greater cytotoxicity than unstimulated NK-92 at this ratio in both cell lines and additionally at the 1:5 ratio in SH-SY5Y cells; no significant reduction in CCK-8 viability was detected in PNT1A or BJ cells at any ratio. Quantitative immunofluorescence analysis demonstrated substantially increased relative fluorescence intensity of all four apoptosis-associated markers in sNK-92-treated PC3 and SH-SY5Y cells compared with their corresponding controls and, in most comparisons, with NK-92-treated cells, whereas changes observed in the PNT1A and BJ non-malignant models examined were markedly smaller. These findings provide preliminary quantitative evidence that anti-CD226-stimulated NK-92 cells exert a preferential cytotoxic effect on the tumor cell models examined, relative to the non-malignant models tested, and induce changes in apoptosis-associated markers consistent with engagement of apoptotic signaling. Further orthogonal validation is warranted.<\/jats:p>","DOI":"10.3390\/ijms27177588","type":"journal-article","created":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T08:00:48Z","timestamp":1787644848000},"page":"7588","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["DNAM-1-Stimulated NK-92 Cells Exert Preferential Cytotoxic and Apoptosis-Associated Effects on Hormone-Independent Solid Tumor Cell Lines"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3882-0728","authenticated-orcid":false,"given":"Mohammadreza","family":"Dastouri","sequence":"first","affiliation":[{"name":"Department of Medical Biology, School of Medicine, Ankara Medipol University, Ankara 06570, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6645-5487","authenticated-orcid":false,"given":"Fatima","family":"Elmusa","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology, Institution of Graduate Schools, Eskisehir Technical University, Eskisehir 26470, T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,25]]},"reference":[{"key":"ref_1","first-page":"209","article-title":"Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA A Cancer J. Clin."},{"key":"ref_2","first-page":"12","article-title":"Cancer statistics, 2024","volume":"74","author":"Siegel","year":"2024","journal-title":"CA A Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/s41572-020-00243-0","article-title":"Prostate cancer","volume":"7","author":"Rebello","year":"2021","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2202","DOI":"10.1056\/NEJMra0804577","article-title":"Recent advances in neuroblastoma","volume":"362","author":"Maris","year":"2010","journal-title":"N. Engl. J. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"16078","DOI":"10.1038\/nrdp.2016.78","article-title":"Neuroblastoma","volume":"2","author":"Matthay","year":"2016","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Ligon, J.A., Wessel, K.M., Shah, N.N., and Glod, J. (2022). Adoptive cell therapy in pediatric and young adult solid tumors: Current status and future directions. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.846346"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.canlet.2013.09.016","article-title":"A personalized view on cancer immunotherapy","volume":"352","author":"Wayteck","year":"2014","journal-title":"Cancer Lett."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.cell.2017.01.017","article-title":"Primary, adaptive, and acquired resistance to cancer immunotherapy","volume":"168","author":"Sharma","year":"2017","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Lian, G., Mak, T.S.-K., Yu, X., and Lan, H.-Y. (2021). Challenges and recent advances in NK cell-targeted immunotherapies in solid tumors. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23010164"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"645","DOI":"10.1038\/nri3044","article-title":"NK cell development, homeostasis and function: Parallels with CD8+ T cells","volume":"11","author":"Sun","year":"2011","journal-title":"Nat. Rev. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1038\/icb.2010.78","article-title":"Activating and inhibitory receptors of natural killer cells","volume":"89","author":"Pegram","year":"2011","journal-title":"Immunol. Cell Biol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4336","DOI":"10.1182\/blood-2006-09-046201","article-title":"Expansion of spleen myeloid suppressor cells represses NK cell cytotoxicity in tumor-bearing host","volume":"109","author":"Liu","year":"2007","journal-title":"Blood"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1038\/nrm2952","article-title":"Mitochondria and cell death: Outer membrane permeabilization and beyond","volume":"11","author":"Tait","year":"2010","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/s41418-017-0012-4","article-title":"Molecular mechanisms of cell death: Recommendations of the Nomenclature Committee on Cell Death 2018","volume":"25","author":"Galluzzi","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/nrc821","article-title":"Targeting death and decoy receptors of the tumour-necrosis factor superfamily","volume":"2","author":"Ashkenazi","year":"2002","journal-title":"Nat. Rev. Cancer"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6969","DOI":"10.1074\/jbc.M113.498253","article-title":"CD226 protein is involved in immune synapse formation and triggers Natural Killer (NK) cell activation via its first extracellular domain","volume":"289","author":"Hou","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1038\/ni.3518","article-title":"Targeting natural killer cells in cancer immunotherapy","volume":"17","author":"Guillerey","year":"2016","journal-title":"Nat. Immunol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1038\/s41577-018-0020-8","article-title":"Regulatory mechanisms in T cell receptor signalling","volume":"18","author":"Gaud","year":"2018","journal-title":"Nat. Rev. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1158\/1078-0432.CCR-12-2065","article-title":"Agonist antibodies to TNFR molecules that costimulate T and NK cells","volume":"19","author":"Melero","year":"2013","journal-title":"Clin. Cancer Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1038\/mt.2014.219","article-title":"Selective inhibition of tumor growth by clonal NK cells expressing an ErbB2\/HER2-specific chimeric antigen receptor","volume":"23","author":"Schonfeld","year":"2015","journal-title":"Mol. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1038\/nri2073","article-title":"Prospects for the use of NK cells in immunotherapy of human cancer","volume":"7","author":"Ljunggren","year":"2007","journal-title":"Nat. Rev. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"625","DOI":"10.1080\/14653240802301872","article-title":"Infusion of the allogeneic cell line NK-92 in patients with advanced renal cell cancer or melanoma: A phase I trial","volume":"10","author":"Arai","year":"2008","journal-title":"Cytotherapy"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhang, C., Oberoi, P., Oelsner, S., Waldmann, A., Lindner, A., Tonn, T., and Wels, W.S. (2017). Chimeric antigen receptor-engineered NK-92 cells: An off-the-shelf cellular therapeutic for targeted elimination of cancer cells and induction of protective antitumor immunity. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.00533"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1987","DOI":"10.1016\/j.ccell.2025.09.013","article-title":"Engineered natural killer cells for cancer therapy","volume":"43","author":"Rezvani","year":"2025","journal-title":"Cancer Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2688610","DOI":"10.1080\/21645515.2026.2688610","article-title":"NK cell-based immunotherapy","volume":"22","author":"Forsberg","year":"2026","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1007\/s12032-023-02080-z","article-title":"The apoptotic effects of NK-92 cells stimulated with an anti-CD226 antibody on MDA-MB-231 triple-negative breast cancer cells","volume":"40","author":"Dastouri","year":"2023","journal-title":"Med. Oncol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1007\/s12032-023-02009-6","article-title":"The effects of KIR2DL4 stimulated NK-92 cells on the apoptotic pathways of HER2+\/HER2\u2212 breast cancer cells","volume":"40","author":"Kilic","year":"2023","journal-title":"Med. Oncol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sayitoglu, E.C., Georgoudaki, A.M., Chrobok, M., Ozkazanc, D., Josey, B.J., Arif, M., Kusser, K., Hartman, M., Chinn, T.M., and Potens, R. (2020). Boosting natural killer cell-mediated targeting of sarcoma through DNAM-1 and NKG2D. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.00040"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"14360","DOI":"10.18632\/oncotarget.3965","article-title":"Highly effective NK cells are associated with good prognosis in patients with metastatic prostate cancer","volume":"6","author":"Pasero","year":"2015","journal-title":"Oncotarget"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9180","DOI":"10.1158\/0008-5472.CAN-04-2682","article-title":"Natural killer cell-mediated killing of freshly isolated neuroblastoma cells: Critical role of DNAX accessory molecule-1-poliovirus receptor interaction","volume":"64","author":"Castriconi","year":"2004","journal-title":"Cancer Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"3901","DOI":"10.1158\/1078-0432.CCR-10-0735","article-title":"Cytotoxicity of activated natural killer cells against pediatric solid tumors","volume":"16","author":"Cho","year":"2010","journal-title":"Clin. Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1553477","DOI":"10.1080\/2162402X.2018.1553477","article-title":"Evaluating natural killer cell cytotoxicity against solid tumors using a microfluidic model","volume":"8","author":"Ayuso","year":"2018","journal-title":"Oncoimmunology"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7588\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T08:02:18Z","timestamp":1787644938000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/17\/7588"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,25]]},"references-count":32,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2026,9]]}},"alternative-id":["ijms27177588"],"URL":"https:\/\/doi.org\/10.3390\/ijms27177588","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,25]]}}}