{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T21:37:19Z","timestamp":1790372239026,"version":"4.1.0"},"reference-count":41,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2021,12,4]],"date-time":"2021-12-04T00:00:00Z","timestamp":1638576000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Science and Technology Innovation Program of Hunan Province","award":["2020RC2051"],"award-info":[{"award-number":["2020RC2051"]}]},{"name":"the The Science and Technology Innovation Program of Hunan Province","award":["2020RC4023"],"award-info":[{"award-number":["2020RC4023"]}]},{"name":"the China Postdoctoral Science Foundation","award":["2020M682576"],"award-info":[{"award-number":["2020M682576"]}]},{"name":"National Nature Sciences Foundation of China","award":["32071262"],"award-info":[{"award-number":["32071262"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>Spider venom is a valuable resource for the development of novel anticancer drugs. In this study, we focused on novel linear amphipathic \u03b1-helical anticancer peptide LVTX-9, which was derived from the cDNA library of the venom gland of the spider Lycosa vittata. The cytotoxicity of LVTX-9 against murine melanoma cells in the range of 1.56\u2013200 \u03bcM was tested and found to be significantly lower than those of most anticancer peptides reported. Its IC50 was determined to be 59.2 \u00b1 19.8 \u03bcM in a serum or 76.3 \u00b1 12.7 \u03bcM in serum-free medium. Fatty acid modification is a promising strategy for improving peptide performance. Therefore, to enhance the cytotoxic activity of LVTX-9, fatty acid modification of this peptide was performed, and five different carbon chain length lipopeptides named LVTX-9-C12-C20 were produced. Among them, the lipopeptide LVTX-9-C18 showed the highest cytotoxic activity in relation to B16-F10 cells, whether in a serum or serum-free medium. Most importantly, the cytotoxic activity of LVTX-9-C18 was improved by about 12.9 times in a serum medium or 19.3 times in a serum-free medium compared to that of LVTX-9. Subsequently, assays including scanning electron microscopy, trypan blue staining, lactate dehydrogenase leakage assay, and hemolytic activity could indicate that the potential direct cell membrane disruption is the main mechanism of LVTX-9-C18 to induce cancer cell death. Furthermore, the LVTX-9-C18 also showed strong cytotoxicity in relation to 3D B16-F10 spheroids, which indicates it might be a promising lead for developing anticancer drugs.<\/jats:p>","DOI":"10.3390\/toxins13120867","type":"journal-article","created":{"date-parts":[[2021,12,7]],"date-time":"2021-12-07T03:21:05Z","timestamp":1638847265000},"page":"867","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Fatty Acid Modification of the Anticancer Peptide LVTX-9 to Enhance Its Cytotoxicity against Malignant Melanoma Cells"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6153-7747","authenticated-orcid":false,"given":"Fengjiao","family":"Li","sequence":"first","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saizhi","family":"Wu","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ninglin","family":"Chen","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyu","family":"Zhu","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinxin","family":"Zhao","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Zhang","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"},{"name":"Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Hunan Normal University), Ministry of Education, College of Chemistry & Chemical Engineering, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youlin","family":"Zeng","sequence":"additional","affiliation":[{"name":"Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Hunan Normal University), Ministry of Education, College of Chemistry & Chemical Engineering, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhonghua","family":"Liu","sequence":"additional","affiliation":[{"name":"The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha 410081, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"145","DOI":"10.3322\/caac.21601","article-title":"Cancer statistics, 2020","volume":"70","author":"Siegel","year":"2020","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3030","DOI":"10.1007\/s00018-006-6315-0","article-title":"Snake venom components and their applications in biomedicine","volume":"63","author":"Koh","year":"2006","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_3","first-page":"1119","article-title":"Animal Venom Peptides: Potential for New Antimicrobial Agents","volume":"17","author":"Macedo","year":"2016","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"219","DOI":"10.3390\/toxins7020219","article-title":"AaeAP1 and AaeAP2: Novel antimicrobial peptides from the venom of the scorpion, Androctonus aeneas: Structural characterisation, molecular cloning of biosynthetic precursor-encoding cDNAs and engineering of analogues with enhanced antimicrobial and anticancer activities","volume":"7","author":"Du","year":"2015","journal-title":"Toxins"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1177\/1535370213513991","article-title":"Scorpion venoms as a potential source of novel cancer therapeutic compounds","volume":"239","author":"Ding","year":"2014","journal-title":"Exp. Biol. Med."},{"key":"ref_6","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_7","doi-asserted-by":"crossref","first-page":"2462","DOI":"10.2174\/138161208785777469","article-title":"Conus venoms\u2014A rich source of peptide-based therapeutics","volume":"14","author":"Han","year":"2008","journal-title":"Curr. Pharm. Des."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1039\/c8tx00022k","article-title":"Anticancer, antimicrobial, and analgesic activities of spider venoms","volume":"7","author":"Akef","year":"2018","journal-title":"Toxicol. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1350","DOI":"10.2174\/156652412803833643","article-title":"A novel spider peptide toxin suppresses tumor growth through dual signaling pathways","volume":"12","author":"Liu","year":"2012","journal-title":"Curr. Mol. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1038\/s41698-020-00129-0","article-title":"Honeybee venom and melittin suppress growth factor receptor activation in HER2-enriched and triple-negative breast cancer","volume":"4","author":"Duffy","year":"2020","journal-title":"NPJ Precis. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9379","DOI":"10.1039\/C7OB02233F","article-title":"Arginine modification of lycosin-I to improve inhibitory activity against cancer cells","volume":"15","author":"Zhang","year":"2017","journal-title":"Org. Biomol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Shuai, Q., Cai, Y., Zhao, G., and Sun, X. (2020). Cell-Penetrating Peptide Modified PEG-PLA Micelles for Efficient PTX Delivery. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21051856"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/S0169-409X(02)00022-4","article-title":"Chemistry for peptide and protein PEGylation","volume":"54","author":"Roberts","year":"2002","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4612","DOI":"10.1021\/acs.molpharmaceut.8b00605","article-title":"The Roles of Fatty-Acid Modification in the Activity of the Anticancer Peptide R-Lycosin-I","volume":"15","author":"Jian","year":"2018","journal-title":"Mol. Pharm."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"C333","DOI":"10.1111\/j.1750-3841.2012.02631.x","article-title":"Improving surface functional properties of tofu whey-derived peptides by chemical modification with fatty acids","volume":"77","author":"Matemu","year":"2012","journal-title":"J. Food Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7857","DOI":"10.1021\/acs.jmedchem.9b00634","article-title":"Monosaccharide Analogues of Anticancer Peptide R-Lycosin-I: Role of Monosaccharide Conjugation in Complexation and the Potential of Lung Cancer Targeting and Therapy","volume":"62","author":"Zhang","year":"2019","journal-title":"J. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8166","DOI":"10.1074\/jbc.M114.621383","article-title":"Stabilization of exosome-targeting peptides via engineered glycosylation","volume":"290","author":"Hung","year":"2015","journal-title":"J. Biol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"782","DOI":"10.1021\/acs.bioconjchem.6b00680","article-title":"Saturated Fatty Acid Analogues of Cell-Penetrating Peptide PepFect14: Role of Fatty Acid Modification in Complexation and Delivery of Splice-Correcting Oligonucleotides","volume":"28","author":"Lehto","year":"2017","journal-title":"Bioconjugate Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7673","DOI":"10.1039\/C5OB00752F","article-title":"Aliphatic acid-conjugated antimicrobial peptides--potential agents with anti-tumor, multidrug resistance-reversing activity and enhanced stability","volume":"13","author":"Deng","year":"2015","journal-title":"Org. Biomol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"111636","DOI":"10.1016\/j.ejmech.2019.111636","article-title":"Design and synthesis of new N-terminal fatty acid modified-antimicrobial peptide analogues with potent in vitro biological activity","volume":"182","author":"Zhong","year":"2019","journal-title":"Eur. J. Med. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11108","DOI":"10.1021\/acs.jmedchem.9b01126","article-title":"Position Effect of Fatty Acid Modification on the Cytotoxicity and Antimetastasis Potential of the Cytotoxic Peptide Lycosin-I","volume":"62","author":"Zhang","year":"2019","journal-title":"J. Med. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1016\/j.toxicon.2015.05.004","article-title":"A survey of the venom of the spider Lycosa vittata by biochemical, pharmacological and transcriptomic analyses","volume":"107","author":"Zhang","year":"2015","journal-title":"Toxicon Off. J. Int. Soc. Toxinol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zhang, P., Yan, Y., Wang, J., Dong, X., Zhang, G., Zeng, Y., and Liu, Z. (2020). An Anti-Cancer Peptide LVTX-8 Inhibits the Proliferation and Migration of Lung Tumor Cells by Regulating Causal Genes\u2019 Expression in p53-Related Pathways. Toxins, 12.","DOI":"10.3390\/toxins12060367"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"73","DOI":"10.2174\/1389557514666141107120954","article-title":"Alpha-helical cationic anticancer peptides: A promising candidate for novel anticancer drugs","volume":"15","author":"Huang","year":"2015","journal-title":"Mini Rev. Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1257","DOI":"10.3389\/fmicb.2019.01257","article-title":"Antimicrobial Peptides: Potential Application in Liver Cancer","volume":"10","author":"Zhang","year":"2019","journal-title":"Front. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.peptides.2014.12.001","article-title":"Cell penetrating peptide TAT can kill cancer cells via membrane disruption after attachment of camptothecin","volume":"63","author":"Song","year":"2015","journal-title":"Peptides"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1016\/j.drudis.2013.05.011","article-title":"Future directions for peptide therapeutics development","volume":"18","author":"Kaspar","year":"2013","journal-title":"Drug Discov. Today"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2700","DOI":"10.1016\/j.bmc.2017.06.052","article-title":"Therapeutic peptides: Historical perspectives, current development trends, and future directions","volume":"26","author":"Lau","year":"2018","journal-title":"Bioorganic Med. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2555","DOI":"10.2174\/1568026619666181221120817","article-title":"Animal Venoms have Potential to Treat Cancer","volume":"18","author":"Chatterjee","year":"2018","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1002\/adhm.201200118","article-title":"Self-assembly of cytotoxic peptide amphiphiles into supramolecular membranes for cancer therapy","volume":"2","author":"Zha","year":"2013","journal-title":"Adv. Healthc. Mater."},{"key":"ref_31","first-page":"151","article-title":"In vitro characterization of the anticancer activity of membrane-active cationic peptides. I. Peptide-mediated cytotoxicity and peptide-enhanced cytotoxic activity of doxorubicin against wild-type and p-glycoprotein over-expressing tumor cell lines","volume":"15","author":"Johnstone","year":"2000","journal-title":"Anti Cancer Drug Des."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"26501","DOI":"10.1021\/acsami.6b08297","article-title":"Surface Physical Activity and Hydrophobicity of Designed Helical Peptide Amphiphiles Control Their Bioactivity and Cell Selectivity","volume":"8","author":"Chen","year":"2016","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_33","first-page":"3062","article-title":"Elevated expression of phosphatidylserine in the outer membrane leaflet of human tumor cells and recognition by activated human blood monocytes","volume":"51","author":"Utsugi","year":"1991","journal-title":"Cancer Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"25","DOI":"10.2174\/1568026615666150703121700","article-title":"Antimicrobial Peptide Structure and Mechanism of Action: A Focus on the Role of Membrane Structure","volume":"16","author":"Lee","year":"2016","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1007\/s13238-014-0061-0","article-title":"Role of helicity of \u03b1-helical antimicrobial peptides to improve specificity","volume":"5","author":"Huang","year":"2014","journal-title":"Protein Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1007\/978-1-4939-2285-7_3","article-title":"Improved methods for classification, prediction, and design of antimicrobial peptides","volume":"1268","author":"Wang","year":"2015","journal-title":"Methods Mol. Biol."},{"key":"ref_37","first-page":"5320","article-title":"Oxygenation and differentiation in multicellular spheroids of human colon carcinoma","volume":"46","author":"Sutherland","year":"1986","journal-title":"Cancer Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.jbiotec.2010.01.012","article-title":"Multicellular tumor spheroids: An underestimated tool is catching up again","volume":"148","author":"Hirschhaeuser","year":"2010","journal-title":"J. Biotechnol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Vinci, M., Gowan, S., Boxall, F., Patterson, L., Zimmermann, M., Court, W., Lomas, C., Mendiola, M., Hardisson, D., and Eccles, S.A. (2012). Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation. BMC Biol., 10.","DOI":"10.1186\/1741-7007-10-29"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.1002\/anie.201005180","article-title":"9-Fluorenylmethoxycarbonyl-based solid-phase synthesis of peptide \u03b1-thioesters","volume":"50","author":"Mende","year":"2011","journal-title":"Angew. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"900","DOI":"10.2174\/15665240113139990045","article-title":"Antimicrobial potential of lycosin-I, a cationic and amphiphilic peptide from the venom of the spider Lycosa singorensis","volume":"13","author":"Tan","year":"2013","journal-title":"Curr. Mol. Med."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/12\/867\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:39:43Z","timestamp":1760168383000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/13\/12\/867"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,4]]},"references-count":41,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2021,12]]}},"alternative-id":["toxins13120867"],"URL":"https:\/\/doi.org\/10.3390\/toxins13120867","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,4]]}}}