The neonatal splice variant of Nav1.5 potentiates in vitro invasive behaviour of MDA-MB-231 human breast cancer cells
Open accessAbstract
Upregulation of functional voltage-gated Na+ channels (VGSCs) occurs in metastatic human breast cancer (BCa) in vitro and in vivo. The present study aimed to ascertain the specific involvement of the "neonatal" splice variant of Nav1.5 (nNav1.5), thought to be predominant, in the VGSC-dependent invasive behaviour of MDA-MB-231 cells. Functional activity of nNav1.5 was suppressed by two different methods targeting nNav1.5: (i) small interfering RNA (siRNA), and (ii) a polyclonal antibody (NESO-pAb); effects upon migration and invasion were determined. nNav1.5 mRNA, protein and signalling were measured using real-time PCR, Western blotting, and patch clamp recording, respectively. Treatment with the siRNA rapidly reduced (by approximately 90%) the level of nNav1.5 (but not adult Nav1.5) mRNA, but the protein reduction was much smaller (approximately 30%), even after 13 days. Nevertheless, the siRNA reduced peak VGSC current density by 33%, and significantly increased the cells' sensitivity to nanomolar tetrodotoxin (TTX). Importantly, the siRNA suppressed in vitro migration by 43%, and eliminated the normally inhibitory effect of TTX. Migrated MDA-MB-231 cells expressed more nNav1.5 protein at the plasma membrane than non-migrated cells. Furthermore, NESO-pAb reduced migration by up to 42%, in a dose-dependent manner. NESO-pAb also reduced Matrigel invasion without affecting proliferation. TTX had no effect on cells already treated with NESO-pAb. It was concluded that nNav1.5 is primarily responsible for the VGSC-dependent enhancement of invasive behaviour in MDA-MB-231 cells. Accordingly, targeting nNav1.5 expression/activity may be useful in clinical management of metastatic BCa.
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- Authors
- Brackenbury, W. J., Chioni, A.-M., Diss, J. K. J., & Djamgoz, M. B. A.
- Affiliations
- Imperial College London (GB)
- University College London (GB)
- Corresponding author
- Djamgoz, Mustafa B. A.
- Venue
- Breast Cancer Research and Treatment
- Published
- 2006-07-13
- Volume
- 101
- Issue
- 2
- Pages
- 149-160
- ISSN
- 0167-6806 (ISSN-L)
- 1573-7217
- PMC
- PMC4122814
- OpenAlex
- W2091102347
- Cited by
- 175
- References
- 69
- Citations by year
- 2026: 4 · 2025: 11 · 2024: 11 · 2023: 10 · 2022: 16 · 2021: 9 · 2020: 11 · 2019: 15 · 2018: 10 · 2017: 5 · 2016: 7 · 2015: 16 · 2014: 7 · 2013: 7 · 2012: 5
- FWCI
- 2.31
- Type
- journal article
- Language
- en
- Open access
- green
- orcid.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Breast Cancer Research and Treatment (publishedVersion)
- PubMed (publishedVersion)
- White Rose Research Online (University of Leeds, The University of Sheffield, University of York) (acceptedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion channel regulation and function
- Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac electrophysiology and arrhythmias
- Keywords
- cardiac sodium channel, voltage-gated sodium channels, breast cancer metastasis, MDA-MB-231 cells, cell invasion, cell migration, tetrodotoxin sensitivity, siRNA knockdown, polyclonal antibodies
- MeSH terms
- Breast Neoplasms, Cell Movement, Female, Humans, Image Processing, Computer-Assisted, Immunohistochemistry, Infant, Newborn, Muscle Proteins, Neoplasm Invasiveness, Tetrodotoxin, Blotting, Western, Sodium Channels, Patch-Clamp Techniques, Microscopy, Confocal, Protein Isoforms, Reverse Transcriptase Polymerase Chain Reaction, Sodium Channel Blockers, RNA, Small Interfering, Cell Line, Tumor, NAV1.5 Voltage-Gated Sodium Channel, In Vitro Techniques
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