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Evaluation of cytotoxicity of isolated fractions from the venom of Iranian cobra snake on skin cancer cell line, B16F10
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Hedieh Farajpoor1    , Delavar Shahbazzadeh2 , Roya Mirzaei 3 |
1- MSc in Toxicology, Department of Toxicology and Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences, Islamic Azad University (IAU), Tehran, Iran
2- 2PhD in Medical Biochemistry, Venom and Bio Therapeutics Molecules Laboratory, Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
3- PhD in Toxicology, Department of Toxicology and Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences, Islamic Azad University (IAU), Tehran, Iran. ,Venom and Bio Therapeutics Molecules Laboratory, Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran , r.mirzaei@student.iautmu.ac.ir |
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Abstract: (918 Views) |
Background: Skin cancer is the most common cancer and very high risk of metastasis of cancer. In recent years, proteins derived from natural sources such as poison animals are considered toxic. The aim of this study was purified Iranian cobra venom using chromatography & fractions obtained by examining toxicity and anti-tumor effect on skin cancer cell line.
Materials and methods: B16F10 melanoma cells in RPMI 1640 medium with 10% FBS were cultured. Iranian cobra venom gel filtration chromatography and ion exchange was purified. Molecular weight fractions were analyzed by SDS-PAGE. The effect of anti-adhesion and cytotoxicity of the active fractions using trypan blue staining and MTT was measured on B16F10 cells were cultured and data were analyzed using student t-test.
Results: Six fractions obtained of gel filtration chromatography. The isolated fractions were evaluated anti-adhesion effect and fraction F3 for further purification by ion exchange chromatography was nominated and 4 fractions were separated. The fraction with a molecular weight of about 47 kDa (F2) was nominated. Totally toxicity was calculated as 49% and 8% on skin cancer cell line and HEK-293, respectively (P≤ 0.05).
Conclusion: In this study, cytotoxic effect on skin cancer cells B16F10 showed that Iranian cobra venom have a protein with highly valued medicinal properties and could be an option for future research against skin cancer.
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| Keywords: Iranian Cobra snake, Chromatography, Skin cancer, MTT assay |
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Full-Text [PDF 419 kb]
(503 Downloads)
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Semi-pilot: Experimental |
Subject:
Pharmacology
Received: 2022/10/8 | Accepted: 2023/01/8 | Published: 2023/05/31
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| References |
1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 2018;68:394-424.
2. Henley SJ, Singh SD, King J, Wilson R, O'Neil ME, Ryerson AB. Invasive cancer incidence and survival-United States, 2011. MMWR Morb Mortal Wkly Rep 2015;64:237.
https://doi.org/10.15585/mmwr.mm6449a1
3. Dubas LE, Ingraffea A. Nonmelanoma skin cancer. Facial Plast Surg Clin North Am 2013;21:43-53.
https://doi.org/10.1016/j.fsc.2012.10.003
4. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. A Cancer Journal for Clinicians 2019; 69:7-34.
https://doi.org/10.3322/caac.21551
5. Tabolacci C, Lentini A, Mattioli P, Provenzano B, Oliverio S, Carlomosti F, et al. Antitumor properties of aloe-emodin and induction of transglutaminase 2 activity in B16-F10 melanoma cells. Life Sci 2010;87:316-24.
https://doi.org/10.1016/j.lfs.2010.07.003
6. Esteva A, Kuprel B, Novoa RA, Ko J, Swetter SM, Blau HM, et al. Dermatologist-level classification of skin cancer with deep neural networks. Nature 2017;542:115.
https://doi.org/10.1038/nature21056
7. DeSantis CE, Lin CC, Mariotto AB, Siegel RL, Stein KD, Kramer JL, et al. Cancer treatment and survivorship statistics, 2014. CA: A Cancer Journal for Clinicians 2014;64:252-71.
https://doi.org/10.3322/caac.21235
8. Institute of Medicine (US) and National Research Council (US) National Cancer Policy Board. Fulfilling the Potential of Cancer Prevention and Early Detection. Curry SJ, Byers T, Hewitt M, editors. Washington (DC): National Academies Press (US); 2003.
9. Simões M, Sousa J, Pais A. Skin cancer and new treatment perspectives: A review. Cancer Lett 2015;357:8-42.
https://doi.org/10.1016/j.canlet.2014.11.001
10. Vyas VK, Brahmbhatt K, Bhatt H, Parmar U. Therapeutic potential of snake venom in cancer therapy: current perspectives. Asian Pac J Trop Biomed 2013;3:156-62.
https://doi.org/10.1016/S2221-1691(13)60042-8
11. Ahn MY, Lee BM, Kim YS. Characterization and cytotoxicity of L-amino acid oxidase from the venom of king cobra (Ophiophagus hannah). Int J Biochem Cell Biol 1997;29:911-9.
https://doi.org/10.1016/S1357-2725(97)00024-1
12. Collares-Buzato CB, Le Sueur LdP, da Cruz-Höfling MA. Impairment of the cell-to-matrix adhesion and cytotoxicity induced by Bothrops moojeni snake venom in cultured renal tubular epithelia. Toxicol Appl Pharmacol 2002;181:124-32.
https://doi.org/10.1006/taap.2002.9404
13. Lucena S, Castro R, Lundin C, Hofstetter A, Alaniz A, Suntravat M, et al. Inhibition of pancreatic tumoral cells by snake venom disintegrins. Toxicon 2015; 93: 136 -43.
https://doi.org/10.1016/j.toxicon.2014.11.228
14. Badr G, Al-Sadoon MK, Rabah DM. Therapeutic efficacy and molecular mechanisms of snake (Walterinnesia aegyptia) venom-loaded silica nanoparticles in the treatment of breast cancer-and prostate cancer-bearing experimental mouse models. Free Radic Biol Med 2013; 65:175-89.
https://doi.org/10.1016/j.freeradbiomed.2013.06.018
15. Abubakar M, Sule M, Pateh U, Abdurahman E, Haruna A, Jahun B. In vitro snake venom detoxifying action of the leaf extract of Guiera senegalensis. J Ethnopharmacol 2000;69:253-7.
https://doi.org/10.1016/S0378-8741(99)00128-2
16. Dubois M, Gilles KA, Hamilton JK, Rebers Pt, Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956;28:350-6.
https://doi.org/10.1021/ac60111a017
17. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680.
https://doi.org/10.1038/227680a0
18. Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. In: Cree IA, Editor. Cancer Cell Culture: Methods and Protocols. Humana Totowa, NJ: Springer; 2011. p. 237-45.
https://doi.org/10.1007/978-1-61779-080-5_20
19. Cree IA. Cancer cell culture: methods and protocols. Humana Totowa, NJ: Springer; 2011.
https://doi.org/10.1007/978-1-61779-080-5
20. Buch K, Peters T, Nawroth T, Sänger M, Schmidberger H, Langguth P. Determination of cell survival after irradiation via clonogenic assay versus multiple MTT Assay-A comparative study. Radiat Oncol 2012;7:1.
https://doi.org/10.1186/1748-717X-7-1
21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
https://doi.org/10.1016/0022-1759(83)90303-4
22. Masters R. Animal cell culture, Cytotoxicity and viability assays. J Immunol Methods 2000; 71:202-3.
23. Shang Z-J, Li Z-B, Li J-R. In vitro effects of nitric oxide synthase inhibitor L-NAME on oral squamous cell carcinoma: a preliminary study. Int J Oral Maxillofac Surg 2006;35:539-43.
https://doi.org/10.1016/j.ijom.2006.01.004
24. Katzung BG. Basic and clinical pharmacology. New York: McGraw-Hill Education; 2017.
25. Rogers HW, Weinstock MA, Feldman SR, Coldiron BM. Incidence estimate of nonmelanoma skin cancer (keratinocyte carcinomas) in the US population, 2012. JAMA Dermatol 2015;151:1081-6.
https://doi.org/10.1001/jamadermatol.2015.1187 [ DOI:10.3322/caac.21492] 2. Leiter U, Garbe C. Epidemiology of melanoma and nonmelanoma skin cancer-the role of sunlight. Adv Exp Med Biol. 2008;624:89-103. [ DOI:10.1007/978-0-387-77574-6_8] 3. Son DJ, Lee JW, Lee YH, Song HS, Lee CK, Hong JT. Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds. Pharmacol Ther 2007;115:246-70. [ DOI:10.1016/j.pharmthera.2007.04.004] 4. Chessum N, Jones K, Pasqua E, Tucker M. Recent advances in cancer therapeutics. Prog Med Chem. 2015;54:1-63. [ DOI:10.1016/bs.pmch.2014.11.002] 5. Thangam R, Gunasekaran P, Kaveri K, Sridevi G, Sundarraj S, Paulpandi M, et al. A novel disintegrin protein from Naja naja venom induces cytotoxicity and apoptosis in human cancer cell lines in vitro. Process Biochemistry 2012;47:1243-9. [ DOI:10.1016/j.procbio.2012.04.020] 6. Torii S, Yamane K, Mashima T, Haga N, Yamamoto K, Fox JW, et al. Molecular cloning and functional analysis of apoxin I, a snake venom-derived apoptosis-inducing factor with L-amino acid oxidase activity. Biochemistry 2000;39:3197-205. [ DOI:10.1021/bi992416z] 7. Zakraoui O, Marcinkiewicz C, Aloui Z, Othman H, Grépin R, Haoues M, et al. Lebein, a snake venom disintegrin, suppresses human colon cancer cells proliferation and tumor‐induced angiogenesis through cell cycle arrest, apoptosis induction and inhibition of VEGF expression. Mol Carcinog 2017;56:18-35. [ DOI:10.1002/mc.22470] 8. Saviola AJ, Burns PD, Mukherjee AK, Mackessy SP. The disintegrin tzabcanin inhibits adhesion and migration in melanoma and lung cancer cells. Int J Biol Macromol 2016; 88:457-64. [ DOI:10.1016/j.ijbiomac.2016.04.008] |
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Farajpoor H, Shahbazzadeh D, Mirzaei R. Evaluation of cytotoxicity of isolated fractions from the venom of Iranian cobra snake on skin cancer cell line, B16F10. MEDICAL SCIENCES 2023; 33 (2) :143-150 URL: http://tmuj.iautmu.ac.ir/article-1-1627-en.html
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