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The neonatal splice variant of Nav1.5 potentiates in vitro invasive behaviour of MDA-MB-231 human breast cancer cells

  • Preclinical study
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Abstract

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 ∼90%) the level of nNav1.5 (but not adult Nav1.5) mRNA, but the protein reduction was much smaller (∼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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Acknowledgements

We would like to thank Dr. J.P. Alao and Prof R.C. Coombes for advice on design of initial RNAi experiments. This study was supported by the Pro Cancer Research Fund (PCRF) and the Bunzl plc - PCRF Programme in Cancer Research.

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Authors and Affiliations

  1. Neuroscience Solutions to Cancer Research Group Division of Cell and Molecular Biology, Imperial College London , Sir Alexander Fleming Building, South Kensington Campus , London , SW7 2AZ, UK

    William J. Brackenbury, Athina-Myrto Chioni & Mustafa B. A. Djamgoz

  2. Medical Molecular Biology Unit, Institute of Child Health, University College, London , WC1N 1EH, UK

    James K. J. Diss

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  1. William J. Brackenbury
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  2. Athina-Myrto Chioni
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  3. James K. J. Diss
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  4. Mustafa B. A. Djamgoz
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Correspondence to Mustafa B. A. Djamgoz.

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Brackenbury, W.J., Chioni, AM., Diss, J.K.J. et al. The neonatal splice variant of Nav1.5 potentiates in vitro invasive behaviour of MDA-MB-231 human breast cancer cells. Breast Cancer Res Treat 101, 149–160 (2007). https://doi.org/10.1007/s10549-006-9281-1

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