{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T16:54:49Z","timestamp":1787504089715,"version":"build-2736575974"},"reference-count":41,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T00:00:00Z","timestamp":1626393600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Financial support from faculty of veterinary medicine, Kasetsart University: FFK. This research was supported in part by the Graduate Program Scholarship from The Graduate School, Kasetsart University, Bangkok, Thailand.","award":["2563"],"award-info":[{"award-number":["2563"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The most common neoplasms in intact female dogs are CMGTs. BmKn-2, an antimicrobial peptide, is derived from scorpion venom and has published anticancer effects in oral and colon human cancer cell lines. Thus, it is highly likely that BmKn-2 could inhibit CMGT cell lines which has not been previously reported. This study investigated the proliferation and apoptotic properties of BmKn-2 via Bax and Bcl-2 relative gene expression in two CMGT cell lines, metastatic (CHMp-5b) and non-metastatic (CHMp-13a). The results showed that BmKn-2 inhibited proliferation and induced apoptosis in the CMGT cell lines. The cell morphology clearly changed and increased apoptosis in a dose dependent of manner. The half maximum inhibitory concentration (IC50) was 30 \u00b5g\/mL for CHMp-5b cell line and 54 \u00b5g\/mL for CHMp-13a cell line. The induction of apoptosis was mediated through Bcl-2 and Bax expression after BmKn-2 treatment. In conclusion, BmKn-2 inhibited proliferation and induced apoptosis in both CHMp-5b and CHMp-13a cell lines via down-regulation of Bcl-2 and up-regulation of Bax relative mRNA expression. Therefore, BmKn-2 could be feasible as candidate treatment for CMGTs.<\/jats:p>","DOI":"10.3390\/ani11072119","type":"journal-article","created":{"date-parts":[[2021,7,16]],"date-time":"2021-07-16T10:52:58Z","timestamp":1626432778000},"page":"2119","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Scorpion Venom Peptide Effects on Inhibiting Proliferation and Inducing Apoptosis in Canine Mammary Gland Tumor Cell Lines"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5650-7802","authenticated-orcid":false,"given":"Kamonporn","family":"Panja","sequence":"first","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkhen Campus, Bangkok 10900, Thailand"},{"name":"Faculty of Veterinary Medicine, Rajamangala University of Technology Tawan-ok, Bangpra, Chonburi 20110, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9971-4318","authenticated-orcid":false,"given":"Supranee","family":"Buranapraditkun","sequence":"additional","affiliation":[{"name":"Division of Allergy and Clinical Immunology, Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand"},{"name":"Center of Excellence in Vaccine Research and Development (Chula Vaccine Research Center-Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand"},{"name":"Thai Pediatric Gastroenterology, Hepatology and Immunology (TPGHAI) Research Unit, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3696-8390","authenticated-orcid":false,"given":"Sittiruk","family":"Roytrakul","sequence":"additional","affiliation":[{"name":"Functional Ingredients and Food Innovation Research Group, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, Pathum Thani 12120, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0769-5234","authenticated-orcid":false,"given":"Attawit","family":"Kovitvadhi","sequence":"additional","affiliation":[{"name":"Department of Physiology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Preeda","family":"Lertwatcharasarakul","sequence":"additional","affiliation":[{"name":"Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kampaeng Saen Campus, Nakhon Pathom 73140, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takayuki","family":"Nakagawa","sequence":"additional","affiliation":[{"name":"Laboratory of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunsumon","family":"Limmanont","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkhen Campus, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3253-8297","authenticated-orcid":false,"given":"Tassanee","family":"Jaroensong","sequence":"additional","affiliation":[{"name":"Department of Companion Animal Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, Bangkhen Campus, Bangkok 10900, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Salas, Y., M\u00e1rquez, A., Diaz, D., and Romero, L. (2015). Epidemiological study of mammary tumors in female dogs diagnosed during the period 2002\u20132012: A growing animal health problem. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0127381"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Thumser-Henner, P., Nytko, K.J., and Bley, C.R. (2020). Mutations of BRCA2 in canine mammary tumors and their targeting potential in clinical therapy. BMC Vet. Res., 16.","DOI":"10.1186\/s12917-020-2247-4"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1136\/vr.103398","article-title":"Adjuvant therapy for highly malignant canine mammary tumours: Cox-2 inhibitor versus chemotherapy: A case\u2013control prospective study","volume":"30","author":"Arenas","year":"2016","journal-title":"Vet. Rec."},{"key":"ref_4","first-page":"1397","article-title":"A cyclized peptide derived from \u03b1 fetoprotein inhibits the proliferation of ER-positive canine mammary cancer cells","volume":"21","author":"Torres","year":"2009","journal-title":"Oncol. Rep."},{"key":"ref_5","first-page":"43","article-title":"Recent advances on canine mammary cancer chemotherapy: A review of studies from 2000 to date","volume":"29","author":"Karayannopoulou","year":"2016","journal-title":"Breast Cancer Res."},{"key":"ref_6","unstructured":"Ettinger, S., and Feldman, E. (1995). Chemotherapy. Textbook of Veterinary Internal Medicine, W. B. Saunders Company, Elsevier, Health Sciences Division 11830 Westline Industrial Drive. [4th ed.]."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1046\/j.1439-0442.2001.00336.x","article-title":"Adjuvant post-operative chemotherapy in bitches with mammary cancer","volume":"48","author":"Karayannopoulou","year":"2001","journal-title":"J. Vet. Med. Ser. A"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1590\/S1678-91992008000300007","article-title":"The venom gland of the scorpion species Euscorpius mingrelicus (Scorpiones: Euscorpiidae): Morphological and ultrastructural characterization","volume":"14","author":"Yigit","year":"2008","journal-title":"J. Venom. Anim. Toxins Incl. Trop. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Liu, Z.-R., and Ji, Y.-H. (2015). Scorpion Venom Research Around the World: Chinese Scorpion Mesobuthus martensii Karsch. Scorpion Venoms Toxinology, 383\u2013385.","DOI":"10.1007\/978-94-007-6404-0_17"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.3389\/fmicb.2018.01159","article-title":"Therapeutic potential of a scorpion venom-derived antimicrobial peptide and its homologs against antibiotic-resistant gram-positive bacteria","volume":"9","author":"Liu","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"294","DOI":"10.3389\/fmicb.2013.00294","article-title":"From antimicrobial to anticancer peptides. A review","volume":"4","author":"Gaspar","year":"2013","journal-title":"Front Microbiol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.1007\/s10989-019-09946-9","article-title":"Antimicrobial peptides (AMPs): Roles, functions and mechanism of action","volume":"26","author":"Seyfi","year":"2020","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Teerapo, K., Roytrakul, S., Sistayanarain, A., and Kunthalert, D. (2019). A scorpion venom peptide derivative BmKn\u201222 with potent antibiofilm activity against Pseudomonas aeruginosa. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0218479"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2807","DOI":"10.7314\/APJCP.2015.16.7.2807","article-title":"BmKn-2 scorpion venom peptide for killing oral cancer cells by apoptosis","volume":"16","author":"Roytrakul","year":"2015","journal-title":"Asian Pac. J. Cancer Prev."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1016\/j.biopha.2016.10.041","article-title":"The scorpion venom peptide BmKn2 induces apoptosis in cancerous but not in normal human oral cells","volume":"84","author":"Satitmanwiwat","year":"2016","journal-title":"Biomed. Pharmacother."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1007\/s10989-014-9417-9","article-title":"Anticancer activities of antimicrobial BmKn2 peptides against oral and colon cancer cells","volume":"20","author":"Arpornsuwan","year":"2014","journal-title":"Int. J. Pept. Res. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.biotechadv.2021.107769","article-title":"Recent developments in animal venom peptide nanotherapeutics with improved selectivity for cancer cells","volume":"50","author":"Giribaldi","year":"2021","journal-title":"Biotechnol. Adv."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"646","DOI":"10.1016\/j.cell.2011.02.013","article-title":"Hallmarks of cancer: The next generation","volume":"144","author":"Hanahan","year":"2011","journal-title":"Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4514","DOI":"10.3892\/mmr.2015.3939","article-title":"Quantitative analysis of the mRNA expression levels of BCL2 and BAX genes in human osteoarthritis and normal articular cartilage: An investigation into their differential expression","volume":"12","author":"Karaliotas","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/nrm2308","article-title":"The BCL-2 protein family: Opposing activities that mediate cell death","volume":"9","author":"Youle","year":"2008","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1111\/vco.12220","article-title":"Cell cycle kinetics, apoptosis rates and gene expressions of MDR-1, TP53, BCL-2 and BAX in transmissible venereal tumour cells and their association with therapy response","volume":"15","author":"Yamatogi","year":"2017","journal-title":"Vet. Comp. Oncol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"235","DOI":"10.4142\/jvs.2016.17.2.235","article-title":"Hematoporphyrin monomethyl ether combined with He-Ne laser irradiation-induced apoptosis in canine breast cancer cells through the mitochondrial pathway","volume":"17","author":"Li","year":"2016","journal-title":"J. Vet. Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1038\/nrc2273","article-title":"Translation of new cancer treatments from pet dogs to humans","volume":"8","author":"Paoloni","year":"2008","journal-title":"Nat. Rev. Cancer"},{"key":"ref_24","first-page":"455","article-title":"Canine mammary tumours as a model to study human breast cancer: Most recent findings","volume":"25","author":"Queiroga","year":"2011","journal-title":"In Vivo"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1016\/j.rvsc.2011.06.012","article-title":"Establishment of a pair of novel cloned tumour cell lines with or without metastatic potential from canine mammary adenocarcinoma","volume":"93","author":"Murai","year":"2012","journal-title":"Res. Vet. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8691569","DOI":"10.1155\/2018\/8691569","article-title":"Zerumbone-loaded nanostructured lipid carrier induces apoptosis of canine mammary adenocarcinoma cells","volume":"2018","author":"Foong","year":"2018","journal-title":"BioMed Res. Int."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2807","DOI":"10.2147\/OTT.S162381","article-title":"5-Azacytidine treatment induces demethylation of DAPK1 and MGMT genes and inhibits growth in canine mammary gland tumor cells","volume":"11","author":"Ren","year":"2018","journal-title":"OncoTargets Ther."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1887","DOI":"10.7150\/thno.16358","article-title":"Targeting negative surface charges of cancer cells by multifunctional nanoprobes","volume":"6","author":"Chen","year":"2016","journal-title":"Theranostics"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/bjc.1972.33","article-title":"Apoptosis: A basic biological phenomenon with wideranging implications in tissue kinetics","volume":"26","author":"Kerr","year":"1972","journal-title":"Br. J. Cancer"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1186\/1756-9966-30-87","article-title":"Apoptosis in cancer: From pathogenesis to treatment","volume":"30","author":"Wong","year":"2011","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1177\/106002809703100113","article-title":"Apoptosis: Its role in the development of malignancies and its potential as a novel therapeutic target","volume":"31","author":"Dixon","year":"1997","journal-title":"Ann. Pharmacother."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1080\/01926230701320337","article-title":"Apoptosis: A review of programmed cell death","volume":"35","author":"Elmore","year":"2007","journal-title":"Toxicol. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1038\/35037722","article-title":"Corpse clearance defines the meaning of cell death","volume":"407","author":"Savill","year":"2000","journal-title":"Nature"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1392","DOI":"10.1016\/j.tiv.2011.04.030","article-title":"Thymoquinone from Nigella sativa was more potent than cisplatin in eliminating of SiHa cells via apoptosis with down-regulation of Bcl-2 protein","volume":"25","author":"Ng","year":"2011","journal-title":"Toxicol. Vitr."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e1264","DOI":"10.1038\/cddis.2014.210","article-title":"Serum-dependent processing of late apoptotic cells for enhanced efferocytosis","volume":"5","author":"Liang","year":"2014","journal-title":"Cell Death Dis."},{"key":"ref_36","first-page":"370","article-title":"Cloning, sequence analysis and structure prediction of B cell lymphoma-2 of Canis familiaris","volume":"3","author":"Mitra","year":"2014","journal-title":"Int. J. Curr. Microbiol. App. Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"846","DOI":"10.1515\/biolog-2015-0094","article-title":"Differential expression of apoptosis-associated genes in canine mammary tumors","volume":"70","author":"Mitra","year":"2015","journal-title":"Biologia"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6306","DOI":"10.1093\/emboj\/20.22.6306","article-title":"Damage-induced Bax N-terminal change, translocation to mitochondria and formation of Bax dimers\/complexes occur regardless of cell fate","volume":"20","author":"Makin","year":"2001","journal-title":"EMBO J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3878","DOI":"10.1093\/emboj\/17.14.3878","article-title":"Enforced dimerization of BAX results in its translocation, mitochondrial dysfunction and apoptosis","volume":"17","author":"Gross","year":"1998","journal-title":"EMBO J."},{"key":"ref_40","first-page":"327","article-title":"Bcl-2\/Bax: A rheostat that regulates an anti-oxidant pathway and cell death","volume":"4","author":"Korsmeyer","year":"1993","journal-title":"Semin. Cancer Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1038\/sj.cdd.4400688","article-title":"Susceptibility to drug-induced apoptosis correlates with differential modulation of Bad, Bcl-2 and Bcl-x L protein levels","volume":"7","author":"Tudor","year":"2000","journal-title":"Cell Death Differ."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/11\/7\/2119\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:30:55Z","timestamp":1760164255000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/11\/7\/2119"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,16]]},"references-count":41,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2021,7]]}},"alternative-id":["ani11072119"],"URL":"https:\/\/doi.org\/10.3390\/ani11072119","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,16]]}}}