{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T18:04:06Z","timestamp":1789495446607,"version":"build-2803163510"},"reference-count":37,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T00:00:00Z","timestamp":1789430400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004156","name":"Mahidol University","doi-asserted-by":"publisher","award":["MU's Strategic Research Fund 2023 (MU-SRF-ST-07A\/66)"],"award-info":[{"award-number":["MU's Strategic Research Fund 2023 (MU-SRF-ST-07A\/66)"]}],"id":[{"id":"10.13039\/501100004156","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016175","name":"Chulabhorn Royal Academy","doi-asserted-by":"publisher","award":["Project code: TNS 2566\/001"],"award-info":[{"award-number":["Project code: TNS 2566\/001"]}],"id":[{"id":"10.13039\/100016175","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>To investigate the anticancer effects of asiatic acid (AA) and asiaticoside (AS) in cisplatin (Cis)-resistant neuroblastoma SH-SY5Y and neural stem cells derived from human stem cells from apical papilla (NSCs-hSCAPs).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec002\">\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>An MTT cell viability assay was performed to determine Cis toxicity and assess the cytotoxic effect of AA or AS on Cis-treated SH-SY5Y and the effect on NSCs-hSCAPs. Subsequently, the combination index (CI) was calculated to observe occurring interactions between the compound and Cis. For mechanistic exploration, flow cytometry using annexin V and PI staining and a caspase-3 activity assay were done to quantify apoptotic cell death. qRT-PCR was used to measure apoptotic and antioxidant mRNA levels. Additionally, the protein levels were analyzed by using the Western blot technique. Finally, to illustrate the putative interactions between the compound and its target, molecular docking was trialled.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec003\">\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      MTT results revealed that AS did not affect both cells. However, 20 \u00b5M AA in combination with 2.5 \u00b5M Cis could significantly reduce SH-SY5Y cell viability with a CI value of 0.41, indicating their synergistic effect. Moreover, the effective concentration of AA did not affect Cis-treated NSCs-hSCAPs, suggesting its low toxicity. Considering SH-SY5Y, AA alone or its combination dramatically increased % apoptotic cells, which showed a minimal increment in Cis alone. Importantly, the combination treatment significantly increased caspase-3 activity. Mechanistically,\n                      <jats:italic>BAX<\/jats:italic>\n                      levels were elevated, whereas\n                      <jats:italic>BCL2, SOD-1<\/jats:italic>\n                      , and\n                      <jats:italic>HO-1<\/jats:italic>\n                      levels were downregulated by AA alone or in combination. These results were also consistent with Western blot analysis. Interestingly, an inhibition of GSK3\u03b2 was observed in the combination treatment, suggesting that these outcomes may be related.The molecular docking illustrated that AA interacted with GSK3\u03b2 and BCL-2 through multiple residues, indicating their potential binding interactions.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec id=\"sec004\">\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>AA and Cis treatment suppressed SH-SY5Y cell viability by enhancing apoptosis, reducing antioxidant gene expression, and potentially inhibiting GSK3\u03b2.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1371\/journal.pone.0358134","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T17:24:36Z","timestamp":1789493076000},"page":"e0358134","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Anticancer activity of asiatic acid in cisplatin-resistant human neuroblastoma SH-SY5Y and its unaffected effect on human neural stem cells"],"prefix":"10.1371","volume":"21","author":[{"given":"Apisada","family":"Jiso","sequence":"first","affiliation":[{"name":"Chiang Mai University","place":["Chiang Mai, Thailand"],"department":["Department of Pharmaceutical Sciences, Faculty of Pharmacy"]},{"name":"Mahidol University","place":["Samut Prakan, Thailand"],"department":["Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laphatrada","family":"Yurasakpong","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Anatomy, Faculty of Science"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amarin","family":"Thongsuk","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Anatomy, Faculty of Science"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6686-5479","authenticated-orcid":true,"given":"Tarinee","family":"Chodchavanchai","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Anatomy, Faculty of Science"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nisarat","family":"Ruangsawasdi","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Pharmacology, Faculty of Dentistry"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Permphan","family":"Dharmasaroja","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Samut Prakan, Thailand"],"department":["Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0827-5207","authenticated-orcid":true,"given":"Anupong Thongklam","family":"Songsaad","sequence":"additional","affiliation":[{"name":"Mahidol University","place":["Bangkok, Thailand"],"department":["Department of Anatomy, Faculty of Dentistry"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-7669-3654","authenticated-orcid":true,"given":"Taweesak","family":"Tangrodchanapong","sequence":"additional","affiliation":[{"name":"Chulabhorn Royal Academy","place":["Bangkok, Thailand"],"department":["School of Radiological Technology, Faculty of Health Science Technology"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,9,15]]},"reference":[{"issue":"2","key":"pone.0358134.ref001","first-page":"204","article-title":"Pediatric neuroblastoma with brain metastasis: the prognostic role of surgery","volume":"32","author":"M Gultekin","year":"2022","journal-title":"Turk Neurosurg"},{"key":"pone.0358134.ref002","doi-asserted-by":"crossref","first-page":"115253","DOI":"10.1016\/j.ejca.2025.115253","article-title":"Survival improvements in patients with melanoma brain metastases and leptomeningeal disease in the modern era: Insights from a nationwide study (2015-2022)","volume":"217","author":"S Pedersen","year":"2025","journal-title":"Eur J Cancer"},{"issue":"2","key":"pone.0358134.ref003","first-page":"317","article-title":"Spinal intradural primary malignant peripheral nerve sheath tumor with leptomeningeal seeding: case report and literature review","volume":"28","author":"H Baharvahdat","year":"2018","journal-title":"Turk Neurosurg"},{"issue":"15","key":"pone.0358134.ref004","doi-asserted-by":"crossref","first-page":"2114","DOI":"10.1016\/S0959-8049(03)00577-X","article-title":"The role of chemotherapy in brain metastases","volume":"39","author":"MJ van den Bent","year":"2003","journal-title":"Eur J Cancer"},{"key":"pone.0358134.ref005","doi-asserted-by":"crossref","first-page":"114010","DOI":"10.1016\/j.expneurol.2022.114010","article-title":"Cisplatin toxicity in the developing brain displays an absolute requirement for caspase-3","volume":"351","author":"KK Hui","year":"2022","journal-title":"Exp Neurol"},{"issue":"6","key":"pone.0358134.ref006","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.1016\/j.molonc.2014.05.009","article-title":"Brain metastasis: new opportunities to tackle therapeutic resistance","volume":"8","author":"J Seoane","year":"2014","journal-title":"Mol Oncol"},{"issue":"1","key":"pone.0358134.ref007","doi-asserted-by":"crossref","first-page":"560","DOI":"10.1186\/s12885-018-4474-7","article-title":"Antiproliferative and apoptotic effect of LY2090314, a GSK-3 inhibitor, in neuroblastoma in vitro","volume":"18","author":"S Kunnimalaiyaan","year":"2018","journal-title":"BMC Cancer"},{"issue":"1","key":"pone.0358134.ref008","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1158\/1535-7163.MCT-20-0497","article-title":"The landscape of glycogen synthase kinase-3 beta genomic alterations in cancer","volume":"20","author":"BA Borden","year":"2021","journal-title":"Mol Cancer Ther"},{"issue":"3","key":"pone.0358134.ref009","first-page":"657","article-title":"Phosphorylation of glycogen synthase kinase-3 beta at serine 9 confers cisplatin resistance in ovarian cancer cells","volume":"31","author":"G Cai","year":"2007","journal-title":"Int J Oncol"},{"issue":"1","key":"pone.0358134.ref010","doi-asserted-by":"crossref","first-page":"7025","DOI":"10.1038\/s41598-024-57624-8","article-title":"Comparative response to PDT with methyl-aminolevulinate and temoporfin in cutaneous and oral squamous cell carcinoma cells","volume":"14","author":"J Nicol\u00e1s-Morala","year":"2024","journal-title":"Sci Rep"},{"key":"pone.0358134.ref011","article-title":"Asiaticoside antagonizes proliferation and chemotherapeutic drug resistance in hepatocellular carcinoma (hcc) cells","volume":"26","author":"Y Ma","year":"2020","journal-title":"Med Sci Monit"},{"issue":"2","key":"pone.0358134.ref012","doi-asserted-by":"crossref","first-page":"184","DOI":"10.3390\/biom10020184","article-title":"Asiatic Acid, extracted from Centella asiatica and induces apoptosis pathway through the phosphorylation p38 mitogen-activated protein kinase in cisplatin-resistant nasopharyngeal carcinoma cells","volume":"10","author":"Y-T Liu","year":"2020","journal-title":"Biomolecules"},{"issue":"5","key":"pone.0358134.ref013","first-page":"4285","article-title":"Effect of asiatic acid on epithelial-mesenchymal transition of human alveolar epithelium A549 cells induced by TGF-\u03b21","volume":"17","author":"Q Cui","year":"2019","journal-title":"Oncol Lett"},{"issue":"1","key":"pone.0358134.ref014","doi-asserted-by":"crossref","first-page":"503","DOI":"10.3892\/mmr.2014.2159","article-title":"Asiaticoside, a component of Centella asiatica, inhibits melanogenesis in B16F10 mouse melanoma","volume":"10","author":"KJ Kwon","year":"2014","journal-title":"Mol Med Rep"},{"issue":"2","key":"pone.0358134.ref015","doi-asserted-by":"crossref","first-page":"698","DOI":"10.1016\/j.xphs.2020.09.015","article-title":"Blood-brain barrier permeability of asiaticoside, madecassoside and asiatic acid in porcine brain endothelial cell model","volume":"110","author":"NA Hanapi","year":"2021","journal-title":"J Pharm Sci"},{"issue":"5","key":"pone.0358134.ref016","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1111\/acer.13368","article-title":"Ethanol, TLR3, and TLR4 agonists have unique innate immune responses in neuron-like SH-SY5Y and microglia-Like BV2","volume":"41","author":"CJ Lawrimore","year":"2017","journal-title":"Alcohol Clin Exp Res"},{"issue":"3","key":"pone.0358134.ref017","doi-asserted-by":"crossref","first-page":"544","DOI":"10.5603\/fm.103887","article-title":"Exploring apoptotic pathways in SH-SY5Y neuroblastoma cells: combined effects of napabucasin and doxorubicin","volume":"84","author":"I \u00dcnl\u00fc","year":"2025","journal-title":"Folia Morphol (Warsz)"},{"issue":"6","key":"pone.0358134.ref018","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0325480","article-title":"Radioprotective effects of asiaticoside and asiatic acid in neural stem cells derived from human stem cells from apical papilla through increasing dose-reduction factor and their lowering effects on SH-SY5Y cell viability","volume":"20","author":"T Tangrodchanapong","year":"2025","journal-title":"PLoS One"},{"key":"pone.0358134.ref019","doi-asserted-by":"crossref","DOI":"10.1590\/1678-7757-2023-0209","article-title":"Characterization of neural stem cells derived from human stem cells from the apical papilla undergoing three-dimensional neurosphere induction","volume":"31","author":"AT Songsaad","year":"2023","journal-title":"J Appl Oral Sci"},{"key":"pone.0358134.ref020","volume-title":"CompuSyn for drug combinations: PC software and user\u2019s guide: a computer program for quantitation of synergism and antagonism in drug combinations, and the determination of IC50 and ED50 and LD50 values","author":"TC Chou","year":"2005"},{"issue":"50","key":"pone.0358134.ref021","doi-asserted-by":"crossref","DOI":"10.3791\/2597","article-title":"Modified annexin V\/propidium iodide apoptosis assay for accurate assessment of cell death","volume":"2011","author":"AM Rieger","year":"2011","journal-title":"J Vis Exp"},{"key":"pone.0358134.ref022","doi-asserted-by":"crossref","first-page":"106552","DOI":"10.1016\/j.archoralbio.2026.106552","article-title":"Differentiation of neural stem cells derived from human stem cells from apical papilla into neuronal-like cells undergoing maturation via 3D-neurospheres formation and neurogenic induction","volume":"185","author":"N Phugdiprapai","year":"2026","journal-title":"Arch Oral Biol"},{"key":"pone.0358134.ref023","doi-asserted-by":"crossref","first-page":"1114295","DOI":"10.3389\/fphar.2023.1114295","article-title":"Therapy resistance in neuroblastoma: mechanisms and reversal strategies","volume":"14","author":"X Zhou","year":"2023","journal-title":"Front Pharmacol"},{"issue":"10","key":"pone.0358134.ref024","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1038\/s41580-023-00629-4","article-title":"Mechanisms of BCL-2 family proteins in mitochondrial apoptosis","volume":"24","author":"PE Czabotar","year":"2023","journal-title":"Nat Rev Mol Cell Biol"},{"issue":"5","key":"pone.0358134.ref025","doi-asserted-by":"crossref","first-page":"118659","DOI":"10.1016\/j.bbamcr.2020.118659","article-title":"GSK-3\u03b2 in DNA repair, apoptosis, and resistance of chemotherapy, radiotherapy of cancer","volume":"1867","author":"J Lin","year":"2020","journal-title":"Biochim Biophys Acta Mol Cell Res"},{"issue":"11","key":"pone.0358134.ref026","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1038\/s41419-019-2081-4","article-title":"Direct impact of cisplatin on mitochondria induces ROS production that dictates cell fate of ovarian cancer cells","volume":"10","author":"M Kleih","year":"2019","journal-title":"Cell Death Dis"},{"key":"pone.0358134.ref027","doi-asserted-by":"crossref","first-page":"1545654","DOI":"10.3389\/fphar.2025.1545654","article-title":"Asiatic acid in anticancer effects: emerging roles and mechanisms","volume":"16","author":"R Chen","year":"2025","journal-title":"Front Pharmacol"},{"issue":"3","key":"pone.0358134.ref028","doi-asserted-by":"crossref","first-page":"515","DOI":"10.3390\/cells10030515","article-title":"Heme oxgenase-1, a cardinal modulator of regulated cell death and inflammation","volume":"10","author":"SW Ryter","year":"2021","journal-title":"Cells"},{"issue":"6","key":"pone.0358134.ref029","doi-asserted-by":"crossref","first-page":"10998","DOI":"10.3390\/ijms140610998","article-title":"Sod1 loss induces intrinsic superoxide accumulation leading to p53-mediated growth arrest and apoptosis","volume":"14","author":"K Watanabe","year":"2013","journal-title":"Int J Mol Sci"},{"issue":"4","key":"pone.0358134.ref030","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.3892\/or.2016.4565","article-title":"GSK-3 inhibitor inhibits cell proliferation and induces apoptosis in human osteosarcoma cells","volume":"35","author":"H Nishimura","year":"2016","journal-title":"Oncol Rep"},{"issue":"4","key":"pone.0358134.ref031","doi-asserted-by":"crossref","first-page":"820","DOI":"10.3390\/cells9040820","article-title":"GSK3: a kinase balancing promotion and resolution of inflammation","volume":"9","author":"L Hoffmeister","year":"2020","journal-title":"Cells"},{"issue":"4","key":"pone.0358134.ref032","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1096\/fj.12-220129","article-title":"Sirt1 inhibits the transcription factor CREB to regulate pituitary growth hormone synthesis","volume":"27","author":"J Monteserin-Garcia","year":"2013","journal-title":"FASEB J"},{"issue":"2","key":"pone.0358134.ref033","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.canlet.2016.07.006","article-title":"GSK-3 inhibition overcomes chemoresistance in human breast cancer","volume":"380","author":"A Ugolkov","year":"2016","journal-title":"Cancer Lett"},{"issue":"6","key":"pone.0358134.ref034","doi-asserted-by":"crossref","first-page":"468","DOI":"10.6026\/97320630016468","article-title":"Molecular docking analysis of Bcl-2 with phyto-compounds","volume":"16","author":"C Kirubhanand","year":"2020","journal-title":"Bioinformation"},{"issue":"7","key":"pone.0358134.ref035","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0180650","article-title":"Asiatic acid protects against cognitive deficits and reductions in cell proliferation and survival in the rat hippocampus caused by 5-fluorouracil chemotherapy","volume":"12","author":"P Chaisawang","year":"2017","journal-title":"PLoS One"},{"issue":"31","key":"pone.0358134.ref036","doi-asserted-by":"crossref","first-page":"22856","DOI":"10.1074\/jbc.M702973200","article-title":"Neural precursor cells are protected from apoptosis induced by trophic factor withdrawal or genotoxic stress by inhibitors of glycogen synthase kinase 3","volume":"282","author":"T-Y Eom","year":"2007","journal-title":"J Biol Chem"},{"issue":"1","key":"pone.0358134.ref037","first-page":"94","article-title":"Selective small-molecule inhibitors of glycogen synthase kinase-3 activity protect primary neurones from death","volume":"77","author":"DA Cross","year":"2001","journal-title":"J Neurochem"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0358134","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T17:24:42Z","timestamp":1789493082000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0358134"}},"subtitle":[],"editor":[{"given":"Irina V.","family":"Lebedeva","sequence":"first","affiliation":[{"name":"Weill Cornell Medicine","place":["UNITED STATES OF AMERICA"]}],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,9,15]]},"references-count":37,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,9,15]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0358134","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,15]]}}}