{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,18]],"date-time":"2026-06-18T13:11:58Z","timestamp":1781788318200,"version":"3.54.5"},"reference-count":49,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2023,3,19]],"date-time":"2023-03-19T00:00:00Z","timestamp":1679184000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"\u201cCompetitiveness, Entrepreneurship &amp; Innovation\u201d (EPAnEK), co-funded by the European Regional Development Fund (ERDF) and national resources","award":["\u201cInTechThrace: Integrated Technologies in biomedical research: multilevel biomarker analysis in Thrace\u201d (MIS Code 5047285)"],"award-info":[{"award-number":["\u201cInTechThrace: Integrated Technologies in biomedical research: multilevel biomarker analysis in Thrace\u201d (MIS Code 5047285)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Aldehyde dehydrogenase 3A1 (ALDH3A1) oxidizes medium-chain aldehydes to their corresponding carboxylic acids. It is expressed at high rates in the human cornea, where it has been characterized as a multi-functional protein displaying various cytoprotective modes of action. Previous studies identified its association with the DNA damage response (DDR) pathway. Here, we utilized a stable transfected HCE-2 (human corneal epithelium) cell line expressing ALDH3A1, to investigate the molecular mechanisms underlying the cytoprotective role(s) of ALDH3A1. Our data revealed morphological differences among the ALDH3A1-expressing and the mock-transfected HCE-2 cells accompanied by differential expression of E-cadherin. Similarly, the ALDH3A1\/HCE-2 cells demonstrated higher mobility, reduced proliferation, upregulation of ZEB1, and downregulation of CDK3, and p57. The expression of ALDH3A1 also affected cell cycle progression by inducing the sequestration of HCE-2 cells at the G2\/M phase. Following 16 h cell treatments with either H2O2 or etoposide, a significantly lower percentage of ALDH3A1\/HCE-2 cells were apoptotic compared to the respective treated mock\/HCE-2 cells. Interestingly, the protective effect of ALDH3A1 expression under these oxidative and genotoxic conditions was accompanied by a reduced formation of \u03b3-H2AX foci and higher levels of total and phospho (Ser15) p53. Finally, ALDH3A1 was found to be localized both in the cytoplasm and the nucleus of transfected HCE-2 cells. Its cellular compartmentalization was not affected by oxidant treatment, while the mechanism by which ALDH3A1 translocates to the nucleus remains unknown. In conclusion, ALDH3A1 protects cells from both apoptosis and DNA damage by interacting with key homeostatic mechanisms associated with cellular morphology, cell cycle, and DDR.<\/jats:p>","DOI":"10.3390\/ijms24065845","type":"journal-article","created":{"date-parts":[[2023,3,20]],"date-time":"2023-03-20T04:33:31Z","timestamp":1679286811000},"page":"5845","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Investigating the Functional Roles of Aldehyde Dehydrogenase 3A1 in Human Corneal Epithelial Cells"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0173-8560","authenticated-orcid":false,"given":"Georgia-Persephoni","family":"Voulgaridou","sequence":"first","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasileios","family":"Theologidis","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Venetikidou","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1607-7470","authenticated-orcid":false,"given":"Ilias","family":"Tsochantaridis","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0139-019X","authenticated-orcid":false,"given":"Avgi","family":"Tsolou","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2255-3426","authenticated-orcid":false,"given":"Maria","family":"Koffa","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1450-3552","authenticated-orcid":false,"given":"Mihalis I.","family":"Panayiotidis","sequence":"additional","affiliation":[{"name":"Department of Cancer Genetics, Therapeutics & Ultrastructural Pathology, The Cyprus Institute of Neurology & Genetics, Ayios Dometios, Nicosia 2371, Cyprus"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0913-4315","authenticated-orcid":false,"given":"Aglaia","family":"Pappa","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, School of Health Sciences, Democritus University of Thrace, 68100 Alexandroupolis, Greece"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1124\/pr.111.005538","article-title":"Aldehyde dehydrogenase inhibitors: A comprehensive review of the pharmacology, mechanism of action, substrate specificity, and clinical application","volume":"64","author":"Koppaka","year":"2012","journal-title":"Pharmacol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"237","DOI":"10.2174\/157488808786734006","article-title":"Aldehyde Dehydrogenase as a Marker for Stem Cells","volume":"3","author":"Moreb","year":"2008","journal-title":"Curr. Stem Cell Res. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1089\/jop.2000.16.173","article-title":"Review: A case for corneal crystallins","volume":"16","author":"Piatigorsky","year":"2000","journal-title":"J. Ocul. Pharmacol. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1016\/0003-9861(89)90465-7","article-title":"Characterization of rat cornea aldehyde dehydrogenase","volume":"274","author":"Evces","year":"1989","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.freeradbiomed.2020.01.183","article-title":"Aldehyde dehydrogenase 3A1 confers oxidative stress resistance accompanied by altered DNA damage response in human corneal epithelial cells","volume":"150","author":"Voulgaridou","year":"2020","journal-title":"Free Radic. Biol. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"25668","DOI":"10.1074\/jbc.M702076200","article-title":"Multiple and additive functions of ALDH3A1 and ALDH1A1: Cataract phenotype and ocular oxidative damage in Aldh3a1(-\/-)\/Aldh1a1(-\/-) knock-out mice","volume":"282","author":"Lassen","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1016\/S0891-5849(03)00070-4","article-title":"ALDH3A1 protects human corneal epithelial cells from ultraviolet- and 4-hydroxy-2-nonenal-induced oxidative damage","volume":"34","author":"Pappa","year":"2003","journal-title":"Free Radic. Biol. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.exer.2006.04.010","article-title":"ALDH3A1: A corneal crystallin with diverse functions","volume":"84","author":"Estey","year":"2007","journal-title":"Exp. Eye Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1042\/bj20030810","article-title":"Human aldehyde dehydrogenase 3A1 (ALDH3A1): Biochemical characterization and immunohistochemical localization in the cornea","volume":"376","author":"Pappa","year":"2003","journal-title":"Biochem. J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1937","DOI":"10.1016\/j.freeradbiomed.2012.02.050","article-title":"Molecular mechanisms of ALDH3A1-mediated cellular protection against 4-hydroxy-2-nonenal","volume":"52","author":"Black","year":"2012","journal-title":"Free Radic. Biol. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.cbi.2016.12.017","article-title":"Corneal haze phenotype in Aldh3a1-null mice: In vivo confocal microscopy and tissue imaging mass spetrometry","volume":"276","author":"Chen","year":"2017","journal-title":"Chem. Biol. Interact."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.biocel.2017.05.017","article-title":"Human aldehyde dehydrogenase 3A1 (ALDH3A1) exhibits chaperone-like function","volume":"89","author":"Voulgaridou","year":"2017","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1459","DOI":"10.1016\/j.freeradbiomed.2006.08.009","article-title":"Antioxidant function of corneal ALDH3A1 in cultured stromal fibroblasts","volume":"41","author":"Lassen","year":"2006","journal-title":"Free Radic. Biol. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"27998","DOI":"10.1074\/jbc.M503698200","article-title":"Human aldehyde dehydrogenase 3A1 inhibits proliferation and promotes survival of human corneal epithelial cells","volume":"280","author":"Pappa","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/BF02789175","article-title":"Fundamentals of flow cytometry","volume":"11","author":"Jaroszeski","year":"1999","journal-title":"Mol. Biotechnol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.biocel.2016.06.004","article-title":"Aldehyde dehydrogenase 3A1 promotes multi-modality resistance and alters gene expression profile in human breast adenocarcinoma MCF-7 cells","volume":"77","author":"Voulgaridou","year":"2016","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"592733","DOI":"10.3389\/fonc.2020.592733","article-title":"Cadherins, Selectins, and Integrins in CAM-DR in Leukemia","volume":"10","author":"Kim","year":"2020","journal-title":"Front. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21590","article-title":"Cancer statistics, 2020","volume":"70","author":"Siegel","year":"2020","journal-title":"CA. Cancer J. Clin."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4769","DOI":"10.1038\/s41388-018-0304-2","article-title":"E-cadherin in contact inhibition and cancer","volume":"37","author":"Mendonsa","year":"2018","journal-title":"Oncogene"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Terzuoli, E., Bellan, C., Aversa, S., Ciccone, V., Morbidelli, L., Giachetti, A., Donnini, S., and Ziche, M. (2019). ALDH3A1 overexpression in melanoma and lung tumors drives cancer stem cell expansion, impairing immune surveillance through enhanced PD-L1 output. Cancers, 11.","DOI":"10.3390\/cancers11121963"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"611","DOI":"10.3892\/ijo.2016.3551","article-title":"Aldehyde dehydrogenase 3A1 is robustly upregulated in gastric cancer stem-like cells and associated with tumorigenesis","volume":"49","author":"Wu","year":"2016","journal-title":"Int. J. Oncol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1080\/15384101.2015.1006048","article-title":"ZEB1: At the crossroads of epithelial-mesenchymal transition, metastasis and therapy resistance","volume":"14","author":"Zhang","year":"2015","journal-title":"Cell Cycle"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"18643","DOI":"10.1074\/jbc.M113.467787","article-title":"Zeb1 Regulates E-cadherin and Epcam (Epithelial Cell Adhesion Molecule) Expression to Control Cell Behavior in Early Zebrafish Development","volume":"288","author":"Vannier","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"321","DOI":"10.15252\/embr.201439333","article-title":"Zeb1 affects epithelial cell adhesion by diverting glycosphingolipid metabolism","volume":"16","author":"Mathow","year":"2015","journal-title":"EMBO Rep."},{"key":"ref_25","first-page":"845","article-title":"Alkali burn causes aldehyde dehydrogenase 3A1 (ALDH3A1) decrease in mouse cornea","volume":"10","author":"Feng","year":"2004","journal-title":"Mol. Vis."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1038\/nrc2602","article-title":"Cell cycle, CDKs and cancer: A changing paradigm","volume":"9","author":"Malumbres","year":"2009","journal-title":"Nat. Rev. Cancer"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.devcel.2008.01.013","article-title":"CDK inhibitors: Cell cycle regulators and beyond","volume":"14","author":"Besson","year":"2008","journal-title":"Dev. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1091","DOI":"10.3389\/fcell.2020.584590","article-title":"Functional Versatility of the CDK Inhibitor p57Kip2","volume":"8","author":"Creff","year":"2020","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2621","DOI":"10.7150\/jca.40171","article-title":"ALDH3A1 acts as a prognostic biomarker and inhibits the epithelial mesenchymal transition of oral squamous cell carcinoma through IL-6\/STAT3 signaling pathway","volume":"11","author":"Qu","year":"2020","journal-title":"J. Cancer"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1514","DOI":"10.1016\/S0959-8049(98)00228-7","article-title":"Etoposide: Four decades of development of a topoisomerase II inhibitor","volume":"34","author":"Hande","year":"1998","journal-title":"Eur. J. Cancer"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1002\/(SICI)1521-1878(199903)21:3<238::AID-BIES8>3.0.CO;2-3","article-title":"Oxidative DNA damage, antioxidants, and cancer","volume":"21","author":"Collins","year":"1999","journal-title":"BioEssays"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6279","DOI":"10.1073\/pnas.1802184115","article-title":"Aldehyde dehydrogenase 3A1 activation prevents radiation-induced xerostomia by protecting salivary stem cells from toxic aldehydes","volume":"115","author":"Saiki","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","first-page":"4777","article-title":"ALDH3A1 Prevents Apoptosis of Corneal Cells Induced by DNA Damaging Agents and Oxidative Stress","volume":"46","author":"Vasiliou","year":"2005","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6109","DOI":"10.1093\/nar\/gkt304","article-title":"Clustered DNA damage induces pan-nuclear H2AX phosphorylation mediated by ATM and DNA\u2013PK","volume":"41","author":"Meyer","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1158\/1078-0432.CCR-12-2529","article-title":"\u03b3-H2AX foci formation as a pharmacodynamic marker of DNA damage produced by DNA cross-linking agents: Results from 2 phase I clinical trials of SJG-136 (SG2000)","volume":"19","author":"Wu","year":"2013","journal-title":"Clin. Cancer Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.freeradbiomed.2013.11.028","article-title":"Aldehyde Dehydrogenase 3a1 Protects Airway Epithelial Cells From Cigarette Smoke-Induced Dna Damage And Cytotoxicity","volume":"68","author":"Jang","year":"2014","journal-title":"Free Radic. Biol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"7666","DOI":"10.1093\/nar\/gku501","article-title":"Critical role for p53-serine 15 phosphorylation in stimulating transactivation at p53-responsive promoters","volume":"42","author":"Loughery","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"a026104","DOI":"10.1101\/cshperspect.a026104","article-title":"The Cell-Cycle Arrest and Apoptotic Functions of p53 in Tumor Initiation and Progression","volume":"6","author":"Chen","year":"2016","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1038\/sj.cdd.4401925","article-title":"The complexity of p53 stabilization and activation","volume":"13","author":"Lavin","year":"2006","journal-title":"Cell Death Differ."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4780","DOI":"10.7150\/jca.58250","article-title":"ALDH3A1 driving tumor metastasis is mediated by p53\/BAG1 in lung adenocarcinoma","volume":"12","author":"Fan","year":"2021","journal-title":"J. Cancer"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Koppaka, V., Chen, Y., Mehta, G., Orlicky, D.J., Thompson, D.C., Jester, J.V., and Vasiliou, V. (2016). ALDH3A1 Plays a Functional Role in Maintenance of Corneal Epithelial Homeostasis. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0146433"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.brainresbull.2009.08.017","article-title":"Corneal aldehyde dehydrogenases: Multiple functions and novel nuclear localization","volume":"81","author":"Stagos","year":"2010","journal-title":"Brain Res. Bull."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1186\/s12964-021-00741-y","article-title":"Types of nuclear localization signals and mechanisms of protein import into the nucleus","volume":"19","author":"Lu","year":"2021","journal-title":"Cell Commun. Signal."},{"key":"ref_44","unstructured":"Linsenmayer, T.F., Beazley, K.E., Cai, C.X., Canner, J.P., Fitch, J.M., Kubilus, J.K., Millholland, J.M., Nurminskaya, M., Talbot, C., and Zak, N.B. (2015). Studies on the Cornea and Lens. Oxidative Stress in Applied Basic Research and Clinical Practice, Humana Press."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1016\/j.bbagen.2010.03.017","article-title":"Nuclear ferritin: A new role for ferritin in cell biology","volume":"1800","author":"Alkhateeb","year":"2010","journal-title":"Biochim. Biophys. Acta Gen. Subj."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"9891","DOI":"10.1073\/pnas.0603779103","article-title":"H2AX phosphorylation within the G1 phase after UV irradiation depends on nucleotide excision repair and not DNA double-strand breaks","volume":"103","author":"Marti","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"3811","DOI":"10.1074\/jbc.M113.514752","article-title":"A missense mutation in Rev7 disrupts formation of Pol\u03b6, impairing mouse development and repair of genotoxic agent-induced DNA lesions","volume":"289","author":"Khalaj","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Voulgaridou, G.P., Mantso, T., Chlichlia, K., Panayiotidis, M.I., and Pappa, A. (2013). Efficient E. coli Expression Strategies for Production of Soluble Human Crystallin ALDH3A1. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0056582"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5845\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:58:44Z","timestamp":1760122724000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/6\/5845"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,19]]},"references-count":49,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2023,3]]}},"alternative-id":["ijms24065845"],"URL":"https:\/\/doi.org\/10.3390\/ijms24065845","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,19]]}}}