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Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro

SIMPLE SUMMARY: Glioblastoma is a brain tumor that is among the deadliest of human cancers. The invasive spreading of the tumor cells allows subpopulations to escape chemo and radiation treatments and infiltrate other areas of the brain where they continue to grow and cause relapses. To achieve thei...

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Autores principales: Varricchio, Alanah, Khan, Sidra, Price, Zoe K., Davis, Rohan A., Ramesh, Sunita A., Yool, Andrea J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9954756/
https://www.ncbi.nlm.nih.gov/pubmed/36831372
http://dx.doi.org/10.3390/cancers15041027
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author Varricchio, Alanah
Khan, Sidra
Price, Zoe K.
Davis, Rohan A.
Ramesh, Sunita A.
Yool, Andrea J.
author_facet Varricchio, Alanah
Khan, Sidra
Price, Zoe K.
Davis, Rohan A.
Ramesh, Sunita A.
Yool, Andrea J.
author_sort Varricchio, Alanah
collection PubMed
description SIMPLE SUMMARY: Glioblastoma is a brain tumor that is among the deadliest of human cancers. The invasive spreading of the tumor cells allows subpopulations to escape chemo and radiation treatments and infiltrate other areas of the brain where they continue to grow and cause relapses. To achieve their high motility, glioblastoma cells appear to harness membrane signaling proteins including aquaporin water channels, glutamate receptors, and ion channels. Work here tested selected combinations of agents that selectively targeted sets of signaling proteins known to be enriched in glioblastomas and showed that blocking both glutamate receptor and aquaporin-1 channels is an attractive means for limiting tumor cell motility without causing cell toxicity that might negatively impact normal brain cell function. Using combinations of agents at threshold doses could enable focusing of the pharmacological effects on cancer cells specifically, minimizing off-target effects. ABSTRACT: Impairing the motility of glioblastoma multiforme (GBM) cells is a compelling goal for new approaches to manage this highly invasive and rapidly lethal human brain cancer. Work here characterized an array of pharmacological inhibitors of membrane ion and water channels, alone and in combination, as tools for restraining glioblastoma spread in human GBM cell lines U87-MG and U251-MG. Aquaporins, AMPA glutamate receptors, and ion channel classes (shown to be upregulated in human GBM at the transcript level and linked to mechanisms of motility in other cell types) were selected as pharmacological targets for analyses. Effective compounds reduced the transwell invasiveness of U87-MG and U251-MG glioblastoma cells by 20–80% as compared with controls, without cytotoxicity. The compounds and doses used were: AqB013 (14 μM); nifedipine (25 µM); amiloride (10 µM); apamin (10 µM); 4-aminopyridine (250 µM); and CNQX (6-cyano-7-nitroquinoxaline-2,3-dione; 30 µM). Invasiveness was quantified in vitro across transwell filter chambers layered with extracellular matrix. Co-application of each of the ion channel agents with the water channel inhibitor AqB013 augmented the inhibition of invasion (20 to 50% greater than either agent alone). The motility impairment achieved by co-application of pharmacological agents differed between the GBM proneural-like subtype U87-MG and classical-like subtype U251-MG, showing patterns consistent with relative levels of target channel expression (Human Protein Atlas database). In addition, two compounds, xanthurenic acid and caelestine C (from the Davis Open Access Natural Product-based Library, Griffith University QLD), were discovered to block invasion at micromolar doses in both GBM lines (IC(50) values from 0.03 to 1 µM), without cytotoxicity, as measured by full mitochondrial activity under conditions matching those in transwell assays and by normal growth in spheroid assays. Mechanisms of action of these agents based on published work are likely to involve modulation of glutamatergic receptor signaling. Treating glioblastoma by the concurrent inhibition of multiple channel targets could be a powerful approach for slowing invasive cell spread without cytotoxic side effects, potentially enhancing the effectiveness of clinical interventions focused on eradicating primary tumors.
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spelling pubmed-99547562023-02-25 Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro Varricchio, Alanah Khan, Sidra Price, Zoe K. Davis, Rohan A. Ramesh, Sunita A. Yool, Andrea J. Cancers (Basel) Article SIMPLE SUMMARY: Glioblastoma is a brain tumor that is among the deadliest of human cancers. The invasive spreading of the tumor cells allows subpopulations to escape chemo and radiation treatments and infiltrate other areas of the brain where they continue to grow and cause relapses. To achieve their high motility, glioblastoma cells appear to harness membrane signaling proteins including aquaporin water channels, glutamate receptors, and ion channels. Work here tested selected combinations of agents that selectively targeted sets of signaling proteins known to be enriched in glioblastomas and showed that blocking both glutamate receptor and aquaporin-1 channels is an attractive means for limiting tumor cell motility without causing cell toxicity that might negatively impact normal brain cell function. Using combinations of agents at threshold doses could enable focusing of the pharmacological effects on cancer cells specifically, minimizing off-target effects. ABSTRACT: Impairing the motility of glioblastoma multiforme (GBM) cells is a compelling goal for new approaches to manage this highly invasive and rapidly lethal human brain cancer. Work here characterized an array of pharmacological inhibitors of membrane ion and water channels, alone and in combination, as tools for restraining glioblastoma spread in human GBM cell lines U87-MG and U251-MG. Aquaporins, AMPA glutamate receptors, and ion channel classes (shown to be upregulated in human GBM at the transcript level and linked to mechanisms of motility in other cell types) were selected as pharmacological targets for analyses. Effective compounds reduced the transwell invasiveness of U87-MG and U251-MG glioblastoma cells by 20–80% as compared with controls, without cytotoxicity. The compounds and doses used were: AqB013 (14 μM); nifedipine (25 µM); amiloride (10 µM); apamin (10 µM); 4-aminopyridine (250 µM); and CNQX (6-cyano-7-nitroquinoxaline-2,3-dione; 30 µM). Invasiveness was quantified in vitro across transwell filter chambers layered with extracellular matrix. Co-application of each of the ion channel agents with the water channel inhibitor AqB013 augmented the inhibition of invasion (20 to 50% greater than either agent alone). The motility impairment achieved by co-application of pharmacological agents differed between the GBM proneural-like subtype U87-MG and classical-like subtype U251-MG, showing patterns consistent with relative levels of target channel expression (Human Protein Atlas database). In addition, two compounds, xanthurenic acid and caelestine C (from the Davis Open Access Natural Product-based Library, Griffith University QLD), were discovered to block invasion at micromolar doses in both GBM lines (IC(50) values from 0.03 to 1 µM), without cytotoxicity, as measured by full mitochondrial activity under conditions matching those in transwell assays and by normal growth in spheroid assays. Mechanisms of action of these agents based on published work are likely to involve modulation of glutamatergic receptor signaling. Treating glioblastoma by the concurrent inhibition of multiple channel targets could be a powerful approach for slowing invasive cell spread without cytotoxic side effects, potentially enhancing the effectiveness of clinical interventions focused on eradicating primary tumors. MDPI 2023-02-06 /pmc/articles/PMC9954756/ /pubmed/36831372 http://dx.doi.org/10.3390/cancers15041027 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Varricchio, Alanah
Khan, Sidra
Price, Zoe K.
Davis, Rohan A.
Ramesh, Sunita A.
Yool, Andrea J.
Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title_full Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title_fullStr Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title_full_unstemmed Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title_short Pharmacological Inhibition of Membrane Signaling Mechanisms Reduces the Invasiveness of U87-MG and U251-MG Glioblastoma Cells In Vitro
title_sort pharmacological inhibition of membrane signaling mechanisms reduces the invasiveness of u87-mg and u251-mg glioblastoma cells in vitro
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9954756/
https://www.ncbi.nlm.nih.gov/pubmed/36831372
http://dx.doi.org/10.3390/cancers15041027
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