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Laboratoire de Physiologie Cellulaire, Institut National de la Santé et de la Recherche Médicale EPI 9938, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq, France
Patch-clamp recordings were used to study ion
currents induced by cell swelling caused by hypotonicity in human
prostate cancer epithelial cells, LNCaP. The reversal potential of the swelling-evoked current suggested that Cl
was the primary
charge carrier (termed ICl,swell). The
selectivity sequence of the underlying volume-regulated anion channels
(VRACs) for different anions was
Br
I > Cl > F > methanesulfonate 
glutamate, with relative
permeability numbers of 1.26, 1.20, 1.0, 0.77, 0.49, and 0.036, respectively. The current-voltage patterns of the whole cell currents
as well as single-channel currents showed moderate outward
rectification. Unitary VRAC conductance was determined at 9.6 ± 1.8 pS. Conventional Cl channel blockers
5-nitro-2-(3-phenylpropylamino)benzoic acid (100 µM) and DIDS (100 µM) inhibited whole cell ICl,swell in a voltage-dependent manner, with the block decreasing from 39.6 ± 9.7% and 71.0 ± 11.0% at +50 mV to 26.2 ± 7.2% and
14.5 ± 6.6% at 
100 mV, respectively. Verapamil (50 µM), a
standard Ca2+ antagonist and P-glycoprotein function
inhibitor, depressed the current by a maximum of 15%. Protein tyrosine
kinase inhibitors downregulated ICl,swell
(genistein with an IC50 of 2.6 µM and lavendustin A by
60 ± 14% at 1 µM). The protein tyrosine phosphatase inhibitor
sodium orthovanadate (500 µM) stimulated
ICl,swell by 54 ± 11%. We conclude that
VRACs in human prostate cancer epithelial cells are modulated via
protein tyrosine phosphorylation.
volume-regulated chloride channels; tyrosine kinase; cell volume
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