{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T06:11:18Z","timestamp":1777011078662,"version":"3.51.4"},"reference-count":52,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2020,1,13]],"date-time":"2020-01-13T00:00:00Z","timestamp":1578873600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI","award":["15K09792"],"award-info":[{"award-number":["15K09792"]}]},{"name":"JSPS KAKENHI","award":["18K09121"],"award-info":[{"award-number":["18K09121"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Allium vegetables such as garlic (Allium sativum L.) are rich in organosulfur compounds that prevent human chronic diseases, including cancer. Of these, diallyl trisulfide (DATS) exhibits anticancer effects against a variety of tumors, including malignant melanoma. Although previous studies have shown that DATS increases intracellular calcium (Ca2+) in different cancer cell types, the role of Ca2+ in the anticancer effect is obscure. In the present study, we investigated the Ca2+ pathways involved in the anti-melanoma effect. We used melittin, the bee venom that can activate a store-operated Ca2+ entry (SOCE) and apoptosis, as a reference. DATS increased apoptosis in human melanoma cell lines in a Ca2+-dependent manner. It also induced mitochondrial Ca2+ (Ca2+mit) overload through intracellular and extracellular Ca2+ fluxes independently of SOCE. Strikingly, acidification augmented Ca2+mit overload, and Ca2+ channel blockers reduced the effect more significantly under acidic pH conditions. On the contrary, acidification mitigated SOCE and Ca2+mit overload caused by melittin. Finally, Ca2+ channel blockers entirely inhibited the anti-melanoma effect of DATS. Our findings suggest that DATS explicitly evokes Ca2+mit overload via a non-SOCE, thereby displaying the anti-melanoma effect.<\/jats:p>","DOI":"10.3390\/ijms21020491","type":"journal-article","created":{"date-parts":[[2020,1,13]],"date-time":"2020-01-13T04:05:51Z","timestamp":1578888351000},"page":"491","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["The Mitochondrial Ca2+ Overload via Voltage-Gated Ca2+ Entry Contributes to an Anti-Melanoma Effect of Diallyl Trisulfide"],"prefix":"10.3390","volume":"21","author":[{"given":"Chinatsu","family":"Nakagawa","sequence":"first","affiliation":[{"name":"Department of Dermatology, Nihon University Hospital, Tokyo 101-830, Japan"},{"name":"Plasma ChemiBio Laboratory, Nasushiobara, Tochigi 329-2813, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Manami","family":"Suzuki-Karasaki","sequence":"additional","affiliation":[{"name":"Plasma ChemiBio Laboratory, Nasushiobara, Tochigi 329-2813, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miki","family":"Suzuki-Karasaki","sequence":"additional","affiliation":[{"name":"Plasma ChemiBio Laboratory, Nasushiobara, Tochigi 329-2813, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Toyoko","family":"Ochiai","sequence":"additional","affiliation":[{"name":"Department of Dermatology, Nihon University Hospital, Tokyo 101-830, Japan"},{"name":"Plasma ChemiBio Laboratory, Nasushiobara, Tochigi 329-2813, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1158-178X","authenticated-orcid":false,"given":"Yoshihiro","family":"Suzuki-Karasaki","sequence":"additional","affiliation":[{"name":"Plasma ChemiBio Laboratory, Nasushiobara, Tochigi 329-2813, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,1,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.biopha.2004.01.004","article-title":"Organosulfur compounds from alliaceae in the prevention of human pathologies","volume":"58","author":"Tapiero","year":"2004","journal-title":"Biomed. Pharmacother."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1021\/jf0106648","article-title":"Physical, chemical, and biological properties of s-allylcysteine, an amino acid derived from garlic","volume":"50","author":"Kodera","year":"2002","journal-title":"J. Agric. Food Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"233","DOI":"10.2174\/18715206113136660370","article-title":"The immunomodulation and anti-inflammatory effects of garlic organosulfur compounds in cancer chemoprevention","volume":"14","author":"Kaschula","year":"2014","journal-title":"Anticancer Agents Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1093\/jnci\/djh123","article-title":"Apoptosis as a novel target for cancer chemoprevention","volume":"96","author":"Sun","year":"2002","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1002\/mnfr.200700527","article-title":"Cancer chemoprevention and mitochondria: Targeting apoptosis in transformed cells via the disruption of mitochondrial bioenergetics\/redox state","volume":"53","author":"Hail","year":"2009","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1063","DOI":"10.1093\/carcin\/4.8.1063","article-title":"Onion and garlic oils inhibit tumor promotion","volume":"4","author":"Belman","year":"1983","journal-title":"Carcinogenesis"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1111\/j.1749-6632.2012.06743.x","article-title":"Molecular mechanisms of garlic-derived allyl sulfides in the inhibition of skin cancer progression","volume":"1271","author":"Wang","year":"2012","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1083","DOI":"10.1002\/cncr.22888","article-title":"Garlic compounds generate reactive oxygen species leading to activation of stress kinases and cysteine proteases for apoptosis in human glioblastoma T98G and U87MG cells","volume":"110","author":"Das","year":"2007","journal-title":"Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1007\/s10495-006-0024-x","article-title":"Garlic compounds induced calpain and intrinsic caspase cascade for apoptosis in human malignant neuroblastoma SH-SY5Y cells","volume":"12","author":"Karmakar","year":"2007","journal-title":"Apoptosis"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/j.canlet.2008.05.027","article-title":"Multitargeted prevention and therapy of cancer by diallyl trisulfide and related Allium vegetable-derived organosulfur compounds","volume":"269","author":"Powolny","year":"2008","journal-title":"Cancer Lett."},{"key":"ref_11","first-page":"743","article-title":"Diallylpolysulfides induce growth arrest and apoptosis","volume":"36","author":"Busch","year":"2010","journal-title":"Int. J. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2328","DOI":"10.1158\/1535-7163.MCT-08-0216","article-title":"Diallyl trisulfide increases the effectiveness of TRAIL and inhibits prostate cancer growth in an orthotopic model: Molecular mechanisms","volume":"7","author":"Shankar","year":"2008","journal-title":"Mol. Cancer Ther."},{"key":"ref_13","first-page":"128","article-title":"Depolarization Controls TRAIL-Sensitization and Tumor-Selective Killing of Cancer Cells: Crosstalk with ROS","volume":"4","author":"Uchida","year":"2014","journal-title":"Front. Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2029","DOI":"10.3892\/ijo.2012.1656","article-title":"Diallyl trisulfide sensitizes human melanoma cells to TRAIL-induced cell death by promoting endoplasmic reticulum-mediated apoptosis","volume":"41","author":"Murai","year":"2012","journal-title":"Int. J. Oncol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4505","DOI":"10.1200\/JCO.2007.13.1011","article-title":"Barriers to effective TRAIL-targeted therapy of malignancy","volume":"25","author":"Dyer","year":"2007","journal-title":"J. Clin. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1038\/onc.2012.164","article-title":"On the TRAIL to successful cancer therapy? Predicting and counteracting resistance against TRAIL-based therapeutics","volume":"32","author":"Dimberg","year":"2013","journal-title":"Oncogene"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"996","DOI":"10.3109\/10715762.2012.690514","article-title":"The three Rs along the TRAIL: Resistance, re-sensitization and reactive oxygen species (ROS)","volume":"46","author":"Mellier","year":"2012","journal-title":"Free Radic. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.freeradbiomed.2013.04.020","article-title":"Mitochondrial superoxide mediates mitochondrial and endoplasmic reticulum dysfunctions in TRAIL-induced apoptosis in Jurkat cells","volume":"61","author":"Inoue","year":"2013","journal-title":"Free Radic. Biol. Med."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"616","DOI":"10.3892\/ijo.2013.2215","article-title":"Crosstalk between mitochondrial ROS and depolarization in the potentiation of TRAIL-induced apoptosis in human tumor cells","volume":"44","author":"Ochiai","year":"2014","journal-title":"Int. J. Oncol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Gualdani, R., de Clippele, M., Ratbi, I., Gailly, P., and Tajeddine, N. (2019). Store-Operated Calcium Entry Contributes to Cisplatin-Induced Cell Death in Non-Small Cell Lung Carcinoma. Cancers, 11.","DOI":"10.3390\/cancers11030430"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1016\/S0014-5793(01)03184-2","article-title":"Delayed oxidant-induced cell death involves activation of phospholipase A2","volume":"509","author":"Zhao","year":"2001","journal-title":"FEBS Lett."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1007\/s10555-011-9339-3","article-title":"Bee venom in cancer therapy","volume":"31","year":"2012","journal-title":"Cancer Metastasis Rev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.lfs.2006.09.024","article-title":"Phospholipase A2-independent Ca2+ entry and subsequent apoptosis induced by melittin in human MG63 osteosarcoma cells","volume":"80","author":"Chu","year":"2007","journal-title":"Life Sci."},{"key":"ref_24","first-page":"939","article-title":"Melittin-induced hyperactivation of phospholipase A2 activity and calcium influx in ras-transformed cells","volume":"8","author":"Sharma","year":"1993","journal-title":"Oncogene"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"11909","DOI":"10.1074\/jbc.M210878200","article-title":"Ca2+-independent phospholipase A2 is a novel determinant of store-operated Ca2+ entry","volume":"278","author":"Smani","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1038\/ncb1089","article-title":"A novel mechanism for the store-operated calcium influx pathway","volume":"6","author":"Smani","year":"2004","journal-title":"Nat. Cell Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3800","DOI":"10.1002\/cam4.1594","article-title":"Vacuolar ATPase as a potential therapeutic target and mediator of treatment resistance in cancer","volume":"7","author":"Whitton","year":"2018","journal-title":"Cancer Med."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1186\/s12943-018-0811-3","article-title":"The curious case of vacuolar ATPase: Regulation of signaling pathways","volume":"17","author":"Pamarthy","year":"2018","journal-title":"Mol. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/0143-4160(94)90091-4","article-title":"Nuclear calcium transport and the role of calcium in apoptosis","volume":"16","author":"Nicotera","year":"1994","journal-title":"Cell Calcium"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1126\/science.281.5381.1309","article-title":"Mitochondria and apoptosis","volume":"281","author":"Green","year":"1998","journal-title":"Science"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1054\/ceca.2000.0138","article-title":"Mitochondrial Ca2+ homeostasis in the regulation of apoptotic and necrotic cell deaths","volume":"28","author":"Zhu","year":"2000","journal-title":"Cell Calcium"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"140","DOI":"10.3389\/fonc.2017.00140","article-title":"IP3 Receptor-Mediated Calcium Signaling and Its Role in Autophagy in Cancer","volume":"7","author":"Kania","year":"2017","journal-title":"Front. Oncol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"180","DOI":"10.3389\/fonc.2017.00180","article-title":"Regulation of Endoplasmic Reticulum-Mitochondria Ca2+ Transfer and Its Importance for Anti-Cancer Therapies","volume":"7","author":"Pedriali","year":"2017","journal-title":"Front. Oncol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1152\/physrev.00004.2005","article-title":"Microdomains of intracellular Ca2+: Molecular determinants and functional consequences","volume":"86","author":"Rizzuto","year":"2006","journal-title":"Physiol. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1417","DOI":"10.1038\/cdd.2016.52","article-title":"The coming of age of the mitochondria-ER contact: A matter of thickness","volume":"23","author":"Giacomello","year":"2016","journal-title":"Cell Death Differ."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1113\/jphysiol.2013.268235","article-title":"The mitochondrial calcium uniporter complex: Molecular components, structure and physiopathological implications","volume":"592","author":"Marchi","year":"2014","journal-title":"J. Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"193","DOI":"10.3892\/ijo.2017.4020","article-title":"Mitochondrial Ca2+ removal amplifies TRAIL cytotoxicity toward apoptosis-resistant tumor cells via promotion of multiple cell death modalities","volume":"51","author":"Takata","year":"2017","journal-title":"Int. J. Oncol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1042\/bj2250413","article-title":"Electroneutral efflux of Ca2+ from liver mitochondria","volume":"225","author":"Brand","year":"1985","journal-title":"Biochem. J."},{"key":"ref_39","first-page":"1699","article-title":"Cyclosporin inhibits mitochondrial calcium efflux in isolated adult rat ventricular cardiomyocytes","volume":"262","author":"Altschuld","year":"1992","journal-title":"Am. J. Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.ceca.2012.03.004","article-title":"The permeability transition pore as a Ca2+ release channel: New answers to an old question","volume":"52","author":"Bernardi","year":"2012","journal-title":"Cell Calcium"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1316","DOI":"10.1016\/j.bbamcr.2011.01.031","article-title":"The mitochondrial permeability transition pore and cyclophilin D in cardioprotection","volume":"1813","author":"Carpi","year":"2011","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_42","first-page":"2041","article-title":"Structural mechanisms of cyclophilin D-dependent control of the mitochondrial permeability transition pore","volume":"1850","author":"Baines","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1146","DOI":"10.3892\/ijo.2017.4096","article-title":"Disrupting mitochondrial Ca2+ homeostasis causes tumor-selective TRAIL sensitization through mitochondrial network abnormalities","volume":"51","author":"Ohshima","year":"2017","journal-title":"Int. J. Oncol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"379","DOI":"10.4161\/chan.24302","article-title":"Emerging roles of store-operated Ca2\u207a entry through STIM and ORAI proteins in immunity, hemostasis, and cancer","volume":"7","author":"Bergmeier","year":"2013","journal-title":"Channels"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1418","DOI":"10.1016\/j.bbamcr.2015.10.003","article-title":"STIM and calcium channel complexes in cancer","volume":"1863","author":"Jardin","year":"2016","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.ceca.2018.07.011","article-title":"ORAI channels and cancer","volume":"74","author":"Chalmers","year":"2018","journal-title":"Cell Calcium"},{"key":"ref_47","first-page":"2944","article-title":"RIP3-mediated STIM1 oligomerization requires intact mitochondrial Ca2+ uptake","volume":"127","author":"Deak","year":"2014","journal-title":"J. Cell Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/s10863-016-9676-6","article-title":"Mitochondrial Ca2+ uptake pathways","volume":"49","author":"Elustondo","year":"2017","journal-title":"J. Bioenerg. Biomembr."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3171","DOI":"10.1111\/j.1742-4658.2007.05848.x","article-title":"Acidic extracellular pH increases calcium influx-triggered phospholipase D activity along with acidic sphingomyelinase activation to induce matrix metalloproteinase-9 expression in mouse metastatic melanoma","volume":"274","author":"Kato","year":"2007","journal-title":"FEBS. J."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1126\/science.1191086","article-title":"The calcium store sensor, STIM1, reciprocally controls Orai and CaV1.2 channels","volume":"330","author":"Wang","year":"2010","journal-title":"Science"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1016\/j.bbamcr.2015.02.014","article-title":"Homer proteins mediate the interaction between STIM1 and Cav1.2 channels","volume":"1853","author":"Dionisio","year":"2015","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"415","DOI":"10.1016\/0092-8674(95)90430-1","article-title":"Decoding of cytosolic calcium oscillations in the mitochondria","volume":"82","author":"Seitz","year":"1995","journal-title":"Cell"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/2\/491\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T13:19:35Z","timestamp":1760361575000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/2\/491"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,1,13]]},"references-count":52,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2020,1]]}},"alternative-id":["ijms21020491"],"URL":"https:\/\/doi.org\/10.3390\/ijms21020491","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,1,13]]}}}