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Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells
BACKGROUND: Spider venom is a potential source of pharmacologically important compounds. Previous studies on spider venoms reported the presence of bioactive molecules that possess cell-modulating activities. Despite these claims, sparse scientific evidence is available on the cytotoxic mechanisms i...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Centro de Estudos de Venenos e Animais Peçonhentos
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7401667/ https://www.ncbi.nlm.nih.gov/pubmed/32788916 http://dx.doi.org/10.1590/1678-9199-JVATITD-2019-0104 |
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author | Mayor, Anna Beatriz R. Guevarra, Leonardo A. Santiago-Bautista, Myla R. Santiago, Librado A. |
author_facet | Mayor, Anna Beatriz R. Guevarra, Leonardo A. Santiago-Bautista, Myla R. Santiago, Librado A. |
author_sort | Mayor, Anna Beatriz R. |
collection | PubMed |
description | BACKGROUND: Spider venom is a potential source of pharmacologically important compounds. Previous studies on spider venoms reported the presence of bioactive molecules that possess cell-modulating activities. Despite these claims, sparse scientific evidence is available on the cytotoxic mechanisms in relation to the components of the spider venom. In this study, we aimed to determine the cytotoxic fractions of the spider venom extracted from Phlogiellus bundokalbo and to ascertain the possible mechanism of toxicity towards human lung adenocarcinoma (A549) cells. METHODS: Spider venom was extracted by electrostimulation. Components of the extracted venom were separated by reversed-phase high performance liquid chromatography (RP-HPLC) using a linear gradient of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 95% acetonitrile (ACN). Cytotoxic activity was evaluated by the MTT assay. Apoptotic or necrotic cell death was assessed by microscopic evaluation in the presence of Hoechst 33342 and Annexin V, Alexa Fluor(TM) 488 conjugate fluorescent stains, and caspase activation assay. Phospholipase A(2) (PLA(2)) activity of the cytotoxic fractions were also measured. RESULTS: We observed and isolated six fractions from the venom of P. bundokalbo collected from Aurora, Zamboanga del Sur. Four of these fractions displayed cytotoxic activities. Fractions AT5-1, AT5-3, and AT5-4 were found to be apoptotic while AT5-6, the least polar among the cytotoxic components, was observed to induce necrosis. PLA(2) activity also showed cytotoxicity in all fractions but presented no relationship between specific activity of PLA(2) and cytotoxicity. CONCLUSION: The venom of P. bundokalbo spider, an endemic tarantula species in the Philippines, contains components that were able to induce either apoptosis or necrosis in A549 cells. |
format | Online Article Text |
id | pubmed-7401667 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Centro de Estudos de Venenos e Animais Peçonhentos |
record_format | MEDLINE/PubMed |
spelling | pubmed-74016672020-08-11 Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells Mayor, Anna Beatriz R. Guevarra, Leonardo A. Santiago-Bautista, Myla R. Santiago, Librado A. J Venom Anim Toxins Incl Trop Dis Research BACKGROUND: Spider venom is a potential source of pharmacologically important compounds. Previous studies on spider venoms reported the presence of bioactive molecules that possess cell-modulating activities. Despite these claims, sparse scientific evidence is available on the cytotoxic mechanisms in relation to the components of the spider venom. In this study, we aimed to determine the cytotoxic fractions of the spider venom extracted from Phlogiellus bundokalbo and to ascertain the possible mechanism of toxicity towards human lung adenocarcinoma (A549) cells. METHODS: Spider venom was extracted by electrostimulation. Components of the extracted venom were separated by reversed-phase high performance liquid chromatography (RP-HPLC) using a linear gradient of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 95% acetonitrile (ACN). Cytotoxic activity was evaluated by the MTT assay. Apoptotic or necrotic cell death was assessed by microscopic evaluation in the presence of Hoechst 33342 and Annexin V, Alexa Fluor(TM) 488 conjugate fluorescent stains, and caspase activation assay. Phospholipase A(2) (PLA(2)) activity of the cytotoxic fractions were also measured. RESULTS: We observed and isolated six fractions from the venom of P. bundokalbo collected from Aurora, Zamboanga del Sur. Four of these fractions displayed cytotoxic activities. Fractions AT5-1, AT5-3, and AT5-4 were found to be apoptotic while AT5-6, the least polar among the cytotoxic components, was observed to induce necrosis. PLA(2) activity also showed cytotoxicity in all fractions but presented no relationship between specific activity of PLA(2) and cytotoxicity. CONCLUSION: The venom of P. bundokalbo spider, an endemic tarantula species in the Philippines, contains components that were able to induce either apoptosis or necrosis in A549 cells. Centro de Estudos de Venenos e Animais Peçonhentos 2020-08-03 /pmc/articles/PMC7401667/ /pubmed/32788916 http://dx.doi.org/10.1590/1678-9199-JVATITD-2019-0104 Text en https://creativecommons.org/licenses/by/4.0/ © The Author(s). 2020 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (https://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Mayor, Anna Beatriz R. Guevarra, Leonardo A. Santiago-Bautista, Myla R. Santiago, Librado A. Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title |
Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title_full |
Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title_fullStr |
Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title_full_unstemmed |
Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title_short |
Phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (A549) cells |
title_sort | phlogiellus bundokalbo spider venom: cytotoxic fractions against human lung adenocarcinoma (a549) cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7401667/ https://www.ncbi.nlm.nih.gov/pubmed/32788916 http://dx.doi.org/10.1590/1678-9199-JVATITD-2019-0104 |
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