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Emerging evidence suggests that vitamin D derivatives may exert anti\u2010cancer effects, including induction of apoptosis, potentially through redox\u2010dependent mechanisms. This study investigated whether vitamin D exposure is associated with reactive oxygen species (ROS) accumulation, mitochondrial dysfunction, and apoptosis in cervical cancer cells. HeLa cells were exposed to physiological, intermediate, and supraphysiological concentrations of 25\u2010hydroxyvitamin D\n                    <jats:sub>3<\/jats:sub>\n                    , as well as calcipotriol, a 1,25\u2010dihydroxyvitamin D analogue. Cell viability assays (crystal violet, Alamar Blue, and Muse cell viability) demonstrated dose\u2010dependent reductions in viability. Flow cytometric analyses demonstrated increased apoptotic markers, evidenced by Annexin V binding and caspase\u20103\/7 activation. The oxidative stress assay showed increased ROS\u2010positive cell populations, accompanied by mitochondrial membrane depolarization. Catalase and total superoxide dismutase activities decreased at the highest effective concentrations. In parallel, decreased total expression of proteins within the MAPK, PI3K\/AKT, and NF\u2010\u03baB pro\u2010survival pathways was observed. These cellular changes were accompanied by apoptotic morphological features visualized by PlasDIC and brightfield microscopy, including membrane blebbing, karyorrhexis, and apoptotic body formation. Collectively, these findings demonstrate concurrent redox disturbance, mitochondrial dysfunction, and apoptosis following supraphysiological vitamin D exposure. NAC co\u2010treatment produced partial attenuation of the ROS\u2010positive population and improved mean viability following calcipotriol treatment, supporting a possible contribution of ROS to calcipotriol\u2010associated cytotoxicity; however, the present experiments do not establish ROS as a necessary mediator of apoptosis.\n                  <\/jats:p>","DOI":"10.1096\/fba.2026-00063","type":"journal-article","created":{"date-parts":[[2026,8,26]],"date-time":"2026-08-26T16:30:58Z","timestamp":1787761858000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Supraphysiological 25\u2010Hydroxyvitamin D\n                    <sub>3<\/sub>\n                    and Calcipotriol Induce Reactive Oxygen Species, Mitochondrial Dysfunction and Apoptosis in\n                    <scp>HeLa<\/scp>\n                    Cervical Cancer Cells"],"prefix":"10.1096","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-8858-8633","authenticated-orcid":false,"given":"Hoi Ching Cherry","family":"Chan","sequence":"first","affiliation":[{"name":"Department of Chemical Pathology, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7050-4974","authenticated-orcid":false,"given":"Sachin","family":"Bhoora","sequence":"additional","affiliation":[{"name":"Vitamin D Cancer Group, Division of Chemical Pathology, Department of Pathology, Faculty of Health Sciences University of Cape Town  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3652-1193","authenticated-orcid":false,"given":"Esther","family":"Zhou","sequence":"additional","affiliation":[{"name":"Vitamin D Cancer Group, Division of Chemical Pathology, Department of Pathology, Faculty of Health Sciences University of Cape Town  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3953-227X","authenticated-orcid":false,"given":"Sumari","family":"Marais","sequence":"additional","affiliation":[{"name":"Department of Physiology, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9982-9710","authenticated-orcid":false,"given":"Tahir S.","family":"Pillay","sequence":"additional","affiliation":[{"name":"Department of Chemical Pathology, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"},{"name":"Tshwane Academic Division National Health Laboratory Services  Pretoria South Africa"},{"name":"Division of Chemical Pathology, Department of Pathology, Faculty of Health Sciences University of Cape Town  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1570-3959","authenticated-orcid":false,"given":"Rivak","family":"Punchoo","sequence":"additional","affiliation":[{"name":"Department of Chemical Pathology, Faculty of Health Sciences University of Pretoria  Pretoria South Africa"},{"name":"Vitamin D Cancer Group, Division of Chemical Pathology, Department of Pathology, Faculty of Health Sciences University of Cape Town  Cape Town South Africa"},{"name":"Division of Chemical Pathology, Department of Pathology, Faculty of Health Sciences University of Cape Town  Cape Town South Africa"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,8,26]]},"reference":[{"key":"e_1_2_11_2_1","unstructured":"Cervical Cancer[WWW Document] 2026 accessed July 25 2026 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