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Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase
TRPM7 is a ubiquitous divalent-selective ion channel with its own kinase domain. Recent studies have shown that suppression of TRPM7 protein expression by RNA interference increases resistance to ischemia-induced neuronal cell death in vivo and in vitro, making the channel a potentially attractive p...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2887440/ https://www.ncbi.nlm.nih.gov/pubmed/20567598 http://dx.doi.org/10.1371/journal.pone.0011161 |
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author | Chen, Hsiang-Chin Xie, Jia Zhang, Zheng Su, Li-Ting Yue, Lixia Runnels, Loren W. |
author_facet | Chen, Hsiang-Chin Xie, Jia Zhang, Zheng Su, Li-Ting Yue, Lixia Runnels, Loren W. |
author_sort | Chen, Hsiang-Chin |
collection | PubMed |
description | TRPM7 is a ubiquitous divalent-selective ion channel with its own kinase domain. Recent studies have shown that suppression of TRPM7 protein expression by RNA interference increases resistance to ischemia-induced neuronal cell death in vivo and in vitro, making the channel a potentially attractive pharmacological target for molecular intervention. Here, we report the identification of the 5-lipoxygenase inhibitors, NDGA, AA861, and MK886, as potent blockers of the TRPM7 channel. Using a cell-based assay, application of these compounds prevented cell rounding caused by overexpression of TRPM7 in HEK-293 cells, whereas inhibitors of 12-lipoxygenase and 15-lipoxygenase did not prevent the change in cell morphology. Application of the 5-lipoxygenase inhibitors blocked heterologously expressed TRPM7 whole-cell currents without affecting the protein's expression level or its cell surface concentration. All three inhibitors were also effective in blocking the native TRPM7 current in HEK-293 cells. However, two other 5-lipoxygenase specific inhibitors, 5,6-dehydro-arachidonic acid and zileuton, were ineffective in suppressing TRPM7 channel activity. Targeted knockdown of 5-lipoxygenase did not reduce TRPM7 whole-cell currents. In addition, application of 5-hydroperoxyeicosatetraenoic acid (5-HPETE), the product of 5-lipoxygenase, or 5-HPETE's downstream metabolites, leukotriene B4 and leukotriene D4, did not stimulate TRPM7 channel activity. These data suggested that NDGA, AA861, and MK886 reduced the TRPM7 channel activity independent of their effect on 5-lipoxygenase activity. Application of AA861 and NDGA reduced cell death for cells overexpressing TRPM7 cultured in low extracellular divalent cations. Moreover, treatment of HEK-293 cells with AA861 increased cell resistance to apoptotic stimuli to a level similar to that obtained for cells in which TRPM7 was knocked down by RNA interference. In conclusion, NDGA, AA861, and MK886 are potent blockers of the TRPM7 channel capable of attenuating TRPM7's function during cell stress, making them effective tools for the biophysical characterization and suppression of TRPM7 channel conductance in vivo. |
format | Text |
id | pubmed-2887440 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-28874402010-06-21 Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase Chen, Hsiang-Chin Xie, Jia Zhang, Zheng Su, Li-Ting Yue, Lixia Runnels, Loren W. PLoS One Research Article TRPM7 is a ubiquitous divalent-selective ion channel with its own kinase domain. Recent studies have shown that suppression of TRPM7 protein expression by RNA interference increases resistance to ischemia-induced neuronal cell death in vivo and in vitro, making the channel a potentially attractive pharmacological target for molecular intervention. Here, we report the identification of the 5-lipoxygenase inhibitors, NDGA, AA861, and MK886, as potent blockers of the TRPM7 channel. Using a cell-based assay, application of these compounds prevented cell rounding caused by overexpression of TRPM7 in HEK-293 cells, whereas inhibitors of 12-lipoxygenase and 15-lipoxygenase did not prevent the change in cell morphology. Application of the 5-lipoxygenase inhibitors blocked heterologously expressed TRPM7 whole-cell currents without affecting the protein's expression level or its cell surface concentration. All three inhibitors were also effective in blocking the native TRPM7 current in HEK-293 cells. However, two other 5-lipoxygenase specific inhibitors, 5,6-dehydro-arachidonic acid and zileuton, were ineffective in suppressing TRPM7 channel activity. Targeted knockdown of 5-lipoxygenase did not reduce TRPM7 whole-cell currents. In addition, application of 5-hydroperoxyeicosatetraenoic acid (5-HPETE), the product of 5-lipoxygenase, or 5-HPETE's downstream metabolites, leukotriene B4 and leukotriene D4, did not stimulate TRPM7 channel activity. These data suggested that NDGA, AA861, and MK886 reduced the TRPM7 channel activity independent of their effect on 5-lipoxygenase activity. Application of AA861 and NDGA reduced cell death for cells overexpressing TRPM7 cultured in low extracellular divalent cations. Moreover, treatment of HEK-293 cells with AA861 increased cell resistance to apoptotic stimuli to a level similar to that obtained for cells in which TRPM7 was knocked down by RNA interference. In conclusion, NDGA, AA861, and MK886 are potent blockers of the TRPM7 channel capable of attenuating TRPM7's function during cell stress, making them effective tools for the biophysical characterization and suppression of TRPM7 channel conductance in vivo. Public Library of Science 2010-06-17 /pmc/articles/PMC2887440/ /pubmed/20567598 http://dx.doi.org/10.1371/journal.pone.0011161 Text en Chen et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Chen, Hsiang-Chin Xie, Jia Zhang, Zheng Su, Li-Ting Yue, Lixia Runnels, Loren W. Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title | Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title_full | Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title_fullStr | Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title_full_unstemmed | Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title_short | Blockade of TRPM7 Channel Activity and Cell Death by Inhibitors of 5-Lipoxygenase |
title_sort | blockade of trpm7 channel activity and cell death by inhibitors of 5-lipoxygenase |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2887440/ https://www.ncbi.nlm.nih.gov/pubmed/20567598 http://dx.doi.org/10.1371/journal.pone.0011161 |
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