Cytotoxic L-Amino Acid Oxidase Isolated from Ophiophagus hannah Induced Apoptosis in Human Melanoma Cell Lines
Keywords:
L-amino acid oxidase, Ophiophagus hannah, apoptosis, human melanomaAbstract
L-amino acid oxidase (LAAO) is one of the components in snake venom that is toxic to cells. There are few studies of anti-melanoma in Thailand. The King cobra, Ophiophagus hannah, is a large and poisonous snake with much venom. Therefore, this study aimed to investigate the cytotoxic potential of LAAO isolated from O. hannah venom in normal human skin fibroblast and human melanoma cell lines. LAAO was isolated by chromatographic procedure based on size exclusion, anion exchanger, and immuno-affinity. MTT assay showed that at 48 h O. hannah LAAO (OH-LAAO) exposure, cell death was rarely found in normal skin fibroblast, whereas approximately 27% death of melanoma cells was exhibited. Phosphati- dyl serine externalization in apoptosis was detected by flow cytometry, which determined the apoptotic cell death induced by OH-LAAO in both cell types. This cell death involved reactive oxygen species, H2O2, generated from the OH-LAAO reaction. The pretreated catalase enzyme could remarkably prevent cell death induced by OH-LAAO. The cytochrome c release into the cytosol and the reduction of Bcl-2 expression indicated that the mitochondrial pathway was considered in the process of apoptosis induced by OH-LAAO. These results provided evidence of OH-LAAO’s potential to be a further therapeutic agent for human melanoma.
References
Singer TP, Kearney EB. The L-amino acid oxidases of snake venom. II. Isolation and characterization of homogenous L-amino acid oxidase. Biochem. 1950; 29: 190-209.
Fung SY, Lee ML, Tan NH. Molecular mechanism of cell death induced by King Cobra (Ophiophagus hannah) venom L-amino acid oxidase. Toxicon. 2015; 96:38–45.
Sakurai Y, Takatsuka H, Yoshioka A, Matsui T, Suzuki M, Titani K, et al. Inhibition of human platelet aggregation by L-amino acid oxidase purified from Naja Naja kaouthia venom. Toxicon. 2001;39(12):1827–33.
Carone SEI, Costa TR, Burin SM, Cintra ACO, Zoccal KF, Bianchini FJ, et al. A new L-amino acid oxidase from Bothrops jararacussu snake venom: Isolation, partial characterization, and assessment of pro-apoptotic and Antiprotozoal Activities. International Journal of Biological Macromolecules. 2017; 103:25–35.
Hiu JJ, Yap MK. Cytotoxicity of snake venom enzymatic toxins: Phospholipase A2 and L-amino acid oxidase. Biochemical Society Transactions. 2020;48(2):719–31.
Zhang L, Wei L-J. ACTX-8, a cytotoxic L-amino acid oxidase isolated from Agkistrodon acutus snake venom, induces apoptosis in Hela cervical cancer cells. Life Sciences. 2007;80(13):1189–97.
Tan CH, Bourges A, Tan KY. King cobra and snakebite envenomation: On the natural history, human-snake relationship and medical importance of Ophiophagus hannah. Journal of Venomous Animals and Toxins including Tropical Diseases. 2021; 27.
Du X-Y, Clemetson KJ. Snake venom L-amino acid oxidases. Toxicon. 2002;40(6):659–65.
Pawelek PD. The structure of L-amino acid oxidase reveals the substrate trajectory into an enantiomerically conserved active site. The EMBO Journal. 2000; 19(16):4204–15.
Lee ML, Tan NH, Fung SY, Sekaran SD. Antibacterial action of a heat-stable form of L-amino acid oxidase isolated from king cobra (Ophiophagus hannah) venom. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology. 2011; 153(2):237–42.
Tan NH, Saifuddin MN. Isolation and characterization of an unusual form of L-amino acid oxidase from King cobra (Ophiophagus hannah) venom. Biochem Int. 1989; 19(4):937–44.
Tan KK, Ler SG, Gunaratne J, Bay BH, Ponnampalam G. In vitro cytotoxicity of L-amino acid oxidase from the venom of Crotalus mitchellii pyrrhus. Toxicon. 2017; 139:20–30.
Fung, Shin YL, Mui LT, Nget H. Proteomic investigation of the molecular mechanism of king cobra venom L-amino acid oxidase induced apoptosis of human breast cancer (MCF-7) cell line. Indian Journal of Experimental Biology. 2018; 56:101-111.
Wang C, Youle RJ. The role of mitochondria in apoptosis. Annual Review of Genetics. 2009;43(1):95–118.
Li Lee M, Chung I, Yee Fung S, Kanthimathi MS, Hong Tan N. Antiproliferative activity of king cobra (Ophiophagus hannah) venom L-amino acid oxidase. Basic & Clinical Pharmacology & Toxicology. 2013;114(4):336–43.
Lee, M.L., Fung, S.Y., Chung, I., Pailoor, J., Cheah, S.H., Tan, N.H. King Cobra (Ophiophagus hannah) Venom L-Amino Acid Oxidase Induces Apoptosis in PC-3 Cells and Suppresses PC-3 Solid Tumor Growth in a Tumor Xenograft Mouse Model. International Journal of Medical Sciences. 2014; 11(6):593-601.
Barrera G. Oxidative stress and lipid peroxidation products in cancer progression and therapy. ISRN Oncology. 2012; 2012:1–21.
Stark G. Functional consequences of oxidative membrane damage. J Membr Biol. 2005; 205(1):1–16.
Morgan MJ, Kim Y-S, Liu Z-G. TNFalpha and reactive oxygen species in necrotic cell death. Cell Res [Internet]. 2008;18(3):343–9.
Costal-Oliveira F, Stransky S, Guerra-Duarte C, Naves de Souza DL, Vivas-Ruiz DE, Yarlequé A, et al. L-amino acid oxidase from Bothrops atrox snake venom triggers autophagy, apoptosis and necrosis in normal human keratinocytes. Sci Rep. 2019; 9(1):781.
Teramoto S, Tomita T, Matsui H, Ohga E, Matsuse T, Ouchi Y. Hydrogen peroxide-induced apoptosis and necrosis in human lung fibroblasts: protective roles of glutathione. Jpn J Pharmacol. 1999; 79(1):33-40.
Garrido C, Galluzzi L, Brunet M, Puig PE, Didelot C, Kroemer G. Mechanisms of cytochrome c release from mitochondria. Cell Death Differ. 2006; 13(9):1423–33.
Izidoro LFM, Sobrinho JC, Mendes MM, Costa TR, Grabner AN, Rodrigues VM, et al. Snake venom L-amino acid oxidases: trends in pharmacology and biochemistry. Biomed Res Int. 2014; 2014:196754.
Opferman JT, Kothari A. Anti-apoptotic BCL-2 family members in development. Cell Death Differ. 2018; 25(1):37–45.
Allsopp TE, Wyatt S, Paterson HF, Davies AM. The proto-oncogene Bcl-2 can selectively rescue neurotrophic factor-dependent neurons from apoptosis. Cell. 1993; 73(2):295–307.
David GB, Isbelle M. Textbook of Modern Surgical Pathology, 2nd ed, Philadelphia, USA. (ISBN: 978-1-4160-3966-2).
Kane DJ, Sarafian TA, Anton R, Hahn H, Gralla EB, Valentine JS, et al. Bcl-2 inhibition of neural death: decreased generation of reactive oxygen species. Science. 1993; 262(5137):1274–7.
Logue SE, Elgendy M, Martin SJ. Expression, purification and use of recombinant annexin V for the detection of apoptotic cells. Nat Protoc. 2009; 4(9):1383–95.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 JOURNAL OF THE MEDICAL TECHNOLOGIST ASSOCIATION OF THAILAND
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
