Cargando…

Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells

Voltage-gated Na(+) channels (VGSCs) are membrane proteins which are normally expressed in excitable cells but have also been detected in cancer cells, where they are thought to be involved in malignancy progression. In this study we examined the ion current and expression profile of VGSC (Na(v)1.5)...

Descripción completa

Detalles Bibliográficos
Autores principales: MOHAMMED, FATIMA H., KHAJAH, MAITHAM A., YANG, MING, BRACKENBURY, WILLIAM J., LUQMANI, YUNUS A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734602/
https://www.ncbi.nlm.nih.gov/pubmed/26718772
http://dx.doi.org/10.3892/ijo.2015.3239
_version_ 1782412939374886912
author MOHAMMED, FATIMA H.
KHAJAH, MAITHAM A.
YANG, MING
BRACKENBURY, WILLIAM J.
LUQMANI, YUNUS A.
author_facet MOHAMMED, FATIMA H.
KHAJAH, MAITHAM A.
YANG, MING
BRACKENBURY, WILLIAM J.
LUQMANI, YUNUS A.
author_sort MOHAMMED, FATIMA H.
collection PubMed
description Voltage-gated Na(+) channels (VGSCs) are membrane proteins which are normally expressed in excitable cells but have also been detected in cancer cells, where they are thought to be involved in malignancy progression. In this study we examined the ion current and expression profile of VGSC (Na(v)1.5) in estrogen receptor (ER)-positive (MCF-7) and silenced (pII) breast cancer cells and its possible influence on their proliferation, motility and invasion. VGSC currents were analysed by whole cell patch clamp recording. Na(v)1.5 expression and localization, in response to EGF stimulation, was examined by western blotting and immunofluorescence respectively. Cell invasion (under-agarose and Matrigel assays), motility (wound healing assay) and proliferation (MTT assay) were assessed in pII cells in response to VGSC blockers, phenytoin (PHT) and tetrodotoxin (TTX), or by siRNA knockdown of Na(v)1.5. The effect of PHT and TTX on modulating EGF-induced phosphorylation of Akt and ERK1/2 was determined by western blotting. Total matrix metalloproteinase (MMP) was determined using a fluorometric-based activity assay. The level of various human proteases was detected by using proteome profiler array kit. VGSC currents were detected in pII cells, but were absent in MCF-7. Na(v)1.5 showed cytoplasmic and perinuclear expression in both MCF-7 and pII cells, with enhanced expression upon EGF stimulation. Treatment of pII cells with PHT, TTX or siRNA significantly reduced invasion towards serum components and EGF, in part through reduction of P-ERK1/2 and proteases such as cathepsin E, kallikrein-10 and MMP-7, as well as total MMP activity. At high concentrations, PHT inhibited motility while TTX reduced cell proliferation. Pharmacological or genetic blockade of Na(v)1.5 may serve as a potential anti-metastatic therapy for breast cancer.
format Online
Article
Text
id pubmed-4734602
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher D.A. Spandidos
record_format MEDLINE/PubMed
spelling pubmed-47346022016-02-18 Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells MOHAMMED, FATIMA H. KHAJAH, MAITHAM A. YANG, MING BRACKENBURY, WILLIAM J. LUQMANI, YUNUS A. Int J Oncol Articles Voltage-gated Na(+) channels (VGSCs) are membrane proteins which are normally expressed in excitable cells but have also been detected in cancer cells, where they are thought to be involved in malignancy progression. In this study we examined the ion current and expression profile of VGSC (Na(v)1.5) in estrogen receptor (ER)-positive (MCF-7) and silenced (pII) breast cancer cells and its possible influence on their proliferation, motility and invasion. VGSC currents were analysed by whole cell patch clamp recording. Na(v)1.5 expression and localization, in response to EGF stimulation, was examined by western blotting and immunofluorescence respectively. Cell invasion (under-agarose and Matrigel assays), motility (wound healing assay) and proliferation (MTT assay) were assessed in pII cells in response to VGSC blockers, phenytoin (PHT) and tetrodotoxin (TTX), or by siRNA knockdown of Na(v)1.5. The effect of PHT and TTX on modulating EGF-induced phosphorylation of Akt and ERK1/2 was determined by western blotting. Total matrix metalloproteinase (MMP) was determined using a fluorometric-based activity assay. The level of various human proteases was detected by using proteome profiler array kit. VGSC currents were detected in pII cells, but were absent in MCF-7. Na(v)1.5 showed cytoplasmic and perinuclear expression in both MCF-7 and pII cells, with enhanced expression upon EGF stimulation. Treatment of pII cells with PHT, TTX or siRNA significantly reduced invasion towards serum components and EGF, in part through reduction of P-ERK1/2 and proteases such as cathepsin E, kallikrein-10 and MMP-7, as well as total MMP activity. At high concentrations, PHT inhibited motility while TTX reduced cell proliferation. Pharmacological or genetic blockade of Na(v)1.5 may serve as a potential anti-metastatic therapy for breast cancer. D.A. Spandidos 2015-11-09 /pmc/articles/PMC4734602/ /pubmed/26718772 http://dx.doi.org/10.3892/ijo.2015.3239 Text en Copyright: © Mohammed et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0) .
spellingShingle Articles
MOHAMMED, FATIMA H.
KHAJAH, MAITHAM A.
YANG, MING
BRACKENBURY, WILLIAM J.
LUQMANI, YUNUS A.
Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title_full Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title_fullStr Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title_full_unstemmed Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title_short Blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
title_sort blockade of voltage-gated sodium channels inhibits invasion of endocrine-resistant breast cancer cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4734602/
https://www.ncbi.nlm.nih.gov/pubmed/26718772
http://dx.doi.org/10.3892/ijo.2015.3239
work_keys_str_mv AT mohammedfatimah blockadeofvoltagegatedsodiumchannelsinhibitsinvasionofendocrineresistantbreastcancercells
AT khajahmaithama blockadeofvoltagegatedsodiumchannelsinhibitsinvasionofendocrineresistantbreastcancercells
AT yangming blockadeofvoltagegatedsodiumchannelsinhibitsinvasionofendocrineresistantbreastcancercells
AT brackenburywilliamj blockadeofvoltagegatedsodiumchannelsinhibitsinvasionofendocrineresistantbreastcancercells
AT luqmaniyunusa blockadeofvoltagegatedsodiumchannelsinhibitsinvasionofendocrineresistantbreastcancercells