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Small-cell Lung Cancer (Human): Potentiation of Endocytic Membrane Activity by Voltage-gated Na+ Channel Expression in Vitro

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Abstract

The possible functional role of voltage-gated Na+ channel (VGSC) expression in controlling endocytic membrane activity in human small-cell lung cancer (SCLC) cell lines (H69, H209, H510) was studied using uptake of horseradish peroxidase (HRP). The normal human airway epithelial (16HBE14o) cell line was used in a comparative approach. Uptake of HRP was vesicular, strongly temperature-sensitive and suppressed by cytoskeletal poisons (cytochalasin D and colchicine), consistent with endocytosis. Compared with the normal cells, HRP uptake into SCLC cells was kinetically more efficient, resulting in more than four-fold higher uptake under optimized conditions. Importantly, HRP uptake into SCLC cells was inhibited significantly by the specific VGSC blocker tetrodotoxin, as well as lidocaine and phenytoin. These effects were dose-dependent. None of these drugs had any effect on the uptake into the 16HBE14o cells. Uptake of HRP into SCLC cells was reduced by ∼66% in Na+-free medium and was partially (∼30%) dependent on extracellular Ca2+. The possibility that the endocytic activity in the H510 SCLC cells involved an endogenous cholinergic system was investigated by testing the effects of carbachol (a cholinergic receptor agonist) and eserine (an inhibitor of acetylcholinesterase). Both drugs inhibited HRP uptake, thereby suggesting that basal cholinergic activity occurred. It is concluded that VGSC upregulation could enhance metastatic cell behavior in SCLC by enhancing endocytic membrane activity.

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Acknowledgements

This study was supported by the Pro Cancer Research Fund - PCRF. We thank Prof Michael J. Seckl for many useful discussions, Drs. Alexander Arcaro and Dieter Gruenert for supplying cell lines, and Prof. Mick Crawley and Mr. Rüstem Önkal for help with statistical analysis.

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  1. Neuroscience Solutions to Cancer Research Group, Department of Biological Sciences, Sir Alexander Fleming Building, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK

    P.U. Onganer & M.B.A. Djamgoz

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  1. P.U. Onganer
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Correspondence to M.B.A. Djamgoz.

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Onganer, P., Djamgoz, M. Small-cell Lung Cancer (Human): Potentiation of Endocytic Membrane Activity by Voltage-gated Na+ Channel Expression in Vitro. J Membrane Biol 204, 67–75 (2005). https://doi.org/10.1007/s00232-005-0747-6

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