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Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer

BACKGROUND: Glioblastoma (GB) cells have the ability to migrate and infiltrate the normal parenchyma, leading to the formation of recurrent tumors often adjacent to the surgical extraction site. We recently showed that Phoneutria nigriventer spider venom (PnV) has anticancer effects mainly on the mi...

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Autores principales: Barreto, Natália, Caballero, Marcus, Bonfanti, Amanda Pires, de Mato, Felipe Cezar Pinheiro, Munhoz, Jaqueline, da Rocha‐e‐Silva, Thomaz A. A., Sutti, Rafael, Vitorino-Araujo, João Luiz, Verinaud, Liana, Rapôso, Catarina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7745393/
https://www.ncbi.nlm.nih.gov/pubmed/33327966
http://dx.doi.org/10.1186/s12935-020-01643-8
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author Barreto, Natália
Caballero, Marcus
Bonfanti, Amanda Pires
de Mato, Felipe Cezar Pinheiro
Munhoz, Jaqueline
da Rocha‐e‐Silva, Thomaz A. A.
Sutti, Rafael
Vitorino-Araujo, João Luiz
Verinaud, Liana
Rapôso, Catarina
author_facet Barreto, Natália
Caballero, Marcus
Bonfanti, Amanda Pires
de Mato, Felipe Cezar Pinheiro
Munhoz, Jaqueline
da Rocha‐e‐Silva, Thomaz A. A.
Sutti, Rafael
Vitorino-Araujo, João Luiz
Verinaud, Liana
Rapôso, Catarina
author_sort Barreto, Natália
collection PubMed
description BACKGROUND: Glioblastoma (GB) cells have the ability to migrate and infiltrate the normal parenchyma, leading to the formation of recurrent tumors often adjacent to the surgical extraction site. We recently showed that Phoneutria nigriventer spider venom (PnV) has anticancer effects mainly on the migration of human GB cell lines (NG97 and U-251). The present work aimed to investigate the effects of isolated components from the venom on migration, invasiveness, morphology and adhesion of GB cells, also evaluating RhoA-ROCK signaling and Na(+)/K(+)-ATPase β2 (AMOG) involvement. METHODS: Human (NG97) GB cells were treated with twelve subfractions (SFs—obtained by HPLC from PnV). Migration and invasion were evaluated by scratch wound healing and transwell assays, respectively. Cell morphology and actin cytoskeleton were shown by GFAP and phalloidin labeling. The assay with fibronectin coated well plate was made to evaluate cell adhesion. Western blotting demonstrated ROCK and AMOG levels and a ROCK inhibitor was used to verify the involvement of this pathway. Values were analyzed by the GraphPad Prism software package and the level of significance was determinate using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test. RESULTS: Two (SF1 and SF11) of twelve SFs, decreased migration and invasion compared to untreated control cells. Both SFs also altered actin cytoskeleton, changed cell morphology and reduced adhesion. SF1 and SF11 increased ROCK expression and the inhibition of this protein abolished the effects of both subfractions on migration, morphology and adhesion (but not on invasion). SF11 also increased Na(+)/K(+)-ATPase β2. CONCLUSION: All components of the venom were evaluated and two SFs were able to impair human glioblastoma cells. The RhoA effector, ROCK, was shown to be involved in the mechanisms of both PnV components. It is possible that AMOG mediates the effect of SF11 on the invasion. Further investigations to isolate and biochemically characterize the molecules are underway.
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spelling pubmed-77453932020-12-18 Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer Barreto, Natália Caballero, Marcus Bonfanti, Amanda Pires de Mato, Felipe Cezar Pinheiro Munhoz, Jaqueline da Rocha‐e‐Silva, Thomaz A. A. Sutti, Rafael Vitorino-Araujo, João Luiz Verinaud, Liana Rapôso, Catarina Cancer Cell Int Primary Research BACKGROUND: Glioblastoma (GB) cells have the ability to migrate and infiltrate the normal parenchyma, leading to the formation of recurrent tumors often adjacent to the surgical extraction site. We recently showed that Phoneutria nigriventer spider venom (PnV) has anticancer effects mainly on the migration of human GB cell lines (NG97 and U-251). The present work aimed to investigate the effects of isolated components from the venom on migration, invasiveness, morphology and adhesion of GB cells, also evaluating RhoA-ROCK signaling and Na(+)/K(+)-ATPase β2 (AMOG) involvement. METHODS: Human (NG97) GB cells were treated with twelve subfractions (SFs—obtained by HPLC from PnV). Migration and invasion were evaluated by scratch wound healing and transwell assays, respectively. Cell morphology and actin cytoskeleton were shown by GFAP and phalloidin labeling. The assay with fibronectin coated well plate was made to evaluate cell adhesion. Western blotting demonstrated ROCK and AMOG levels and a ROCK inhibitor was used to verify the involvement of this pathway. Values were analyzed by the GraphPad Prism software package and the level of significance was determinate using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test. RESULTS: Two (SF1 and SF11) of twelve SFs, decreased migration and invasion compared to untreated control cells. Both SFs also altered actin cytoskeleton, changed cell morphology and reduced adhesion. SF1 and SF11 increased ROCK expression and the inhibition of this protein abolished the effects of both subfractions on migration, morphology and adhesion (but not on invasion). SF11 also increased Na(+)/K(+)-ATPase β2. CONCLUSION: All components of the venom were evaluated and two SFs were able to impair human glioblastoma cells. The RhoA effector, ROCK, was shown to be involved in the mechanisms of both PnV components. It is possible that AMOG mediates the effect of SF11 on the invasion. Further investigations to isolate and biochemically characterize the molecules are underway. BioMed Central 2020-12-17 /pmc/articles/PMC7745393/ /pubmed/33327966 http://dx.doi.org/10.1186/s12935-020-01643-8 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Primary Research
Barreto, Natália
Caballero, Marcus
Bonfanti, Amanda Pires
de Mato, Felipe Cezar Pinheiro
Munhoz, Jaqueline
da Rocha‐e‐Silva, Thomaz A. A.
Sutti, Rafael
Vitorino-Araujo, João Luiz
Verinaud, Liana
Rapôso, Catarina
Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title_full Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title_fullStr Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title_full_unstemmed Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title_short Spider venom components decrease glioblastoma cell migration and invasion through RhoA-ROCK and Na(+)/K(+)-ATPase β2: potential molecular entities to treat invasive brain cancer
title_sort spider venom components decrease glioblastoma cell migration and invasion through rhoa-rock and na(+)/k(+)-atpase β2: potential molecular entities to treat invasive brain cancer
topic Primary Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7745393/
https://www.ncbi.nlm.nih.gov/pubmed/33327966
http://dx.doi.org/10.1186/s12935-020-01643-8
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