{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T04:57:10Z","timestamp":1783573030105,"version":"3.55.0"},"reference-count":42,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2020,11,5]],"date-time":"2020-11-05T00:00:00Z","timestamp":1604534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Since the current treatments have not resulted in the desired outcomes for melanoma patients, there is a need to identify more effective medications. Together with other snake venom proteins, cytotoxin-II has shown promising results in tumoral cells. In this study, recombinant cytotoxin-II (rCTII) was expressed in SHuffle\u00ae T7 Express cells, while the epitope mapping of rCTII was performed to reveal the antibody-binding regions of rCTII. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to assess the viability of SK-MEL-3 and HFF-2 cells after treating these cells with rCTII. The qRT-PCR was performed to evaluate the expression levels of matrix metallopeptidase 3 (MMP-3), SMAD2, SMAD3, caspase-8, caspase-9, and miR-214 in order to reveal the rCTII-induced signaling pathways in melanoma. Our results have shown that two regions of amino acids, 6\u201316 and 19\u201344, as predicted epitopes of this toxin, are essential for understanding the toxicity of rCTII. Treating the melanoma cells with rCTII substantially inhibited the transforming growth factor-beta (TGF-\u03b2)\u2013SMAD signaling pathway and down-regulated the expression of MMP-3 and miR-214 as well. This cytotoxin also restored apoptosis mainly via the intrinsic pathway. The down-regulation of MMP-3 and miR-214 might be associated with the anti-metastatic property of rCTII in melanoma. The inhibitory effect of rCTII on the TGF-\u03b2 signaling pathway might be associated with increased apoptosis and decreased cancer cell proliferation. It is interesting to see that the IC50 value of rCTII has been lower in the melanoma cells than non-tumoral cells, which may indicate its potential effects as a drug. In conclusion, rCTII, as a novel medication, might serve as a potent and efficient anticancer drug in melanoma.<\/jats:p>","DOI":"10.3390\/molecules25215148","type":"journal-article","created":{"date-parts":[[2020,11,5]],"date-time":"2020-11-05T09:04:34Z","timestamp":1604567074000},"page":"5148","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Targeting TGF-\u03b2-Mediated SMAD Signaling Pathway via Novel Recombinant Cytotoxin II: A Potent Protein from Naja naja oxiana Venom in Melanoma"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3243-233X","authenticated-orcid":false,"given":"Afshin","family":"Derakhshani","sequence":"first","affiliation":[{"name":"Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran"},{"name":"IRCCS Istituto Tumori \u201cGiovanni Paolo II\u201d of Bari, 70124 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7814-7318","authenticated-orcid":false,"given":"Nicola","family":"Silvestris","sequence":"additional","affiliation":[{"name":"IRCCS Istituto Tumori \u201cGiovanni Paolo II\u201d of Bari, 70124 Bari, Italy"},{"name":"Department of Biomedical Sciences and Human Oncology (DIMO), University of Bari, 70124 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6432-9647","authenticated-orcid":false,"given":"Nima","family":"Hemmat","sequence":"additional","affiliation":[{"name":"Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zahra","family":"Asadzadeh","sequence":"additional","affiliation":[{"name":"Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4865-8779","authenticated-orcid":false,"given":"Mahdi","family":"Abdoli Shadbad","sequence":"additional","affiliation":[{"name":"Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Niloufar Sadat","family":"Nourbakhsh","sequence":"additional","affiliation":[{"name":"Department of Biology, Islamic Azad University, Varamin-Pishva Branch, Varamin, Tehran 33817-74895, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3470-0278","authenticated-orcid":false,"given":"Leila","family":"Mobasheri","sequence":"additional","affiliation":[{"name":"Department of Immunology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand 97178-53577, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parviz","family":"Vahedi","sequence":"additional","affiliation":[{"name":"Department of Anatomical Sciences, Maragheh University of Medical Sciences, Maragheh 51656-65931, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morteza","family":"Shahmirzaie","sequence":"additional","affiliation":[{"name":"Pharmaceutical Sciences Research Center, Shahid Beheshti University of Medical Sciences, Niayesh Highway, Valiasr Ave, Tehran 19919-53381, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7014-6828","authenticated-orcid":false,"given":"Oronzo","family":"Brunetti","sequence":"additional","affiliation":[{"name":"IRCCS Istituto Tumori \u201cGiovanni Paolo II\u201d of Bari, 70124 Bari, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9883-9186","authenticated-orcid":false,"given":"Hossein","family":"Safarpour","sequence":"additional","affiliation":[{"name":"Cellular &amp; Molecular Research Center, Birjand University of Medical Sciences, Birjand 97178-53577, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8642-6795","authenticated-orcid":false,"given":"Behzad","family":"Baradaran","sequence":"additional","affiliation":[{"name":"Immunology Research Center, Tabriz University of Medical Sciences, Tabriz 51656-65811, Iran"},{"name":"Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz 51666-14766, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,11,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1007\/s11912-018-0732-8","article-title":"The world of melanoma: Epidemiologic, genetic, and anatomic differences of melanoma across the globe","volume":"20","author":"Dimitriou","year":"2018","journal-title":"Curr. 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