{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T22:46:23Z","timestamp":1791413183834,"version":"4.3.3"},"reference-count":37,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2015,9,18]],"date-time":"2015-09-18T00:00:00Z","timestamp":1442534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>This study aims to explore the effects and mechanisms of hepcidin, a potential antimicrobial peptide from Tilapia, and epirubicin (Epi), an antineoplastic agent, on the generation of reactive oxygen species (ROS) and link the ROS levels to the reversal mechanisms of multidrug resistance (MDR) by epirubicin and hepcidin in human squamous cell carcinoma SCC15 and human embryonal carcinoma NT2D1 cells. The cells, pretreated with hepcidin, epirubicin, or a combination of these compounds in PEGylated liposomes, were used to validate the molecular mechanisms involved in inhibiting efflux transporters and inducing apoptosis as evaluated by cytotoxicity, intracellular accumulation, mRNA levels, cell cycle distribution, and caspase activity of this combination. We found that hepcidin significantly enhanced the cytotoxicity of epirubicin in liposomes. The  co-incubation of epirubicin with hepcidin in liposomes intensified the ROS production, including hydrogen peroxide and superoxide free radicals. Hepcidin significantly increased epirubicin intracellular uptake into NT2D1 and SCC15 cells, as supported by the diminished mRNA expressions of MDR1, MDR-associated protein (MRP) 1, and MRP2. Hepcidin and\/or epirubicin in liposomes triggered apoptosis, as verified by the reduced mitochondrial membrane potential, increased sub-G1 phase of cell cycle, incremental populations of apoptosis using annexin V\/PI assay, and chromatin condensation. As far as we know, this is the first example showing that PEGylated liposomal TH1-5 and epirubicin gives rise to cell death in human squamous carcinoma and testicular embryonic carcinoma cells through the reduced epirubicin efflux via ROS-mediated suppression of P-gp and MRPs and concomitant initiation of mitochondrial apoptosis pathway. Hence, hepcidin in PEGylated liposomes may function as an adjuvant to anticancer drugs, thus demonstrating a novel strategy for reversing MDR.<\/jats:p>","DOI":"10.3390\/ijms160922711","type":"journal-article","created":{"date-parts":[[2015,9,21]],"date-time":"2015-09-21T02:25:40Z","timestamp":1442802340000},"page":"22711-22734","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["The Use of a Liposomal Formulation Incorporating an Antimicrobial Peptide from Tilapia as a New Adjuvant to Epirubicin in Human Squamous Cell Carcinoma and Pluripotent Testicular Embryonic Carcinoma Cells"],"prefix":"10.3390","volume":"16","author":[{"given":"Yu-Li","family":"Lo","sequence":"first","affiliation":[{"name":"Department and Institute of Pharmacology, National Yang-Ming University, No. 155, Sec. 2,  Linong Street, Taipei 112, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hsin-Pin","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences and Technology, National University of Tainan, Tainan 700, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei-Chen","family":"Tu","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences and Technology, National University of Tainan, Tainan 700, Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2015,9,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.fsi.2014.10.040","article-title":"Transcriptome analysis of medaka following epinecidin-1 and TH1\u20135 treatment of NNV infection","volume":"42","author":"Wang","year":"2015","journal-title":"Fish Shellfish Immunol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3627","DOI":"10.1016\/j.biomaterials.2014.01.032","article-title":"The mechanisms by which pardaxin, a natural cationic antimicrobial peptide, targets the endoplasmic reticulum and induces c-FOS","volume":"35","author":"Ting","year":"2014","journal-title":"Biomaterials"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1552","DOI":"10.1016\/j.biomaterials.2013.10.082","article-title":"Molecular mechanisms of anticancer action and cell selectivity of short alpha-helical peptides","volume":"35","author":"Chen","year":"2014","journal-title":"Biomaterials"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1186\/1471-2407-11-116","article-title":"Small lytic peptides escape the inhibitory effect of heparan sulfate on the surface of cancer cells","volume":"11","author":"Fadnes","year":"2011","journal-title":"BMC Cancer"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2290","DOI":"10.18632\/oncotarget.2959","article-title":"The antimicrobial peptide pardaxin exerts potent anti-tumor activity against canine perianal gland adenoma","volume":"6","author":"Pan","year":"2015","journal-title":"Oncotarget"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"151","DOI":"10.3390\/ph8020151","article-title":"Reactive oxygen species, apoptosis, antimicrobial peptides and human inflammatory diseases","volume":"8","author":"Oyinloye","year":"2015","journal-title":"Pharmaceuticals"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/1471-2490-8-5","article-title":"Antimicrobial peptides of the Cecropin-family show potent antitumor activity against bladder cancer cells","volume":"8","author":"Suttmann","year":"2008","journal-title":"BMC Urol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1922","DOI":"10.1016\/j.molimm.2006.09.031","article-title":"Three different hepcidins from tilapia, Oreochromis mossambicus: Analysis of their expressions and biological functions","volume":"44","author":"Huang","year":"2007","journal-title":"Mol. 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