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Predominant expression of Kv1.3 voltage-gated K+ channel subunit in rat prostate cancer cell lines: electrophysiological, pharmacological and molecular characterisation

  • Ion Channels, Transporters
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Abstract

Voltage-gated K+ currents expressed in two rat prostate cancer ('Dunning') cell lines of markedly different metastatic ability were characterised using electrophysiological, pharmacological and molecular approaches. Whole-cell patch-clamp recordings showed that both strongly metastatic MAT-LyLu and weakly metastatic AT-2 cell lines possessed outward (delayed-rectifier type) K+ currents, which activated at around −40 mV. From the parameters measured, several characteristics of the two cell lines were similar. However, a number of statistically significant differences were noted for MAT-LyLu versus the AT-2 cells as follows: (1) current densities were smaller; (2) the slope factor for channel activation was smaller; (3) the voltage at which current was half-inactivated, and the slope factor for channel inactivation were greater; (4) the time constants for current decay at −20 and 0 mV were smaller; and (5) the residual peak current was larger following 60 s of repetitive voltage pulses for stimulation frequencies in the range 0.05–0.2 Hz. On the other hand, the K+ currents in both cell lines showed similar pharmacological profiles. Thus, the currents were blocked by 4-aminopyridine, tetraethylammonium, verapamil, margatoxin, and charybdotoxin, with highly similar IC50s for given blockers. The electrophysiological and pharmacological data taken together suggested expression of voltage-gated K+ channels of the Kv1 family, expression of the Kv1.3 subunit being predominant. Western blot and RT-PCR tests both confirmed that the cells indeed expressed Kv1.3 and to a lesser extent Kv1.4 and Kv1.6 channel α-subunits. In view of the similarity of channel expression in the two cell lines, voltage-gated K+ channel activity may not be a primary determinant of metastatic potential in the rat model of prostate cancer, but the possible contribution of K+ channel activity to the metastatic process is discussed.

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Acknowledgements

This work was supported by grants from the AICR (Association for International Cancer Research), KANCATAK and the MRC. We gratefully acknowledge the support of contributors to the Prostate Cancer Research Fund (PCRF). In addition, we thank Prof. James Trimmer for the Kv1.5 antibody, Prof. Clare Isacke for providing the secondary antibodies, Dr. N. Mohammed for help with preparation of the Western blot figure, and Dr. Jo Hirano for help with the PCR experiments.

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  1. Department of Biological Sciences, Imperial College of Science, Technology and Medicine, Sir Alexander Fleming Building, London , SW7 2AZ, UK

    S. P. Fraser, J. A. Grimes, J. K. J. Diss, D. Stewart, J. O. Dolly & M. B. A. Djamgoz

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  1. S. P. Fraser
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Correspondence to S. P. Fraser.

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Fraser, S.P., Grimes, J.A., Diss, J.K.J. et al. Predominant expression of Kv1.3 voltage-gated K+ channel subunit in rat prostate cancer cell lines: electrophysiological, pharmacological and molecular characterisation. Pflugers Arch - Eur J Physiol 446, 559–571 (2003). https://doi.org/10.1007/s00424-003-1077-0

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