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  • Changes in the motility of B16F10 melanoma cells induced by alterations in resting calcium influx.

Changes in the motility of B16F10 melanoma cells induced by alterations in resting calcium influx.

Melanoma research (2002-07-26)
J L Cox, T Lancaster, C G Carlson
ABSTRACT

Alterations in the extracellular Ca(2+) or K(+) concentration had significant influences on the motility of B16F10 melanoma cells measured in the absence of exogenous integrins using a conventional Boyden chamber assay. At normal K(+) concentrations, motility increased slightly when the concentration of Ca(2+) was increased 10-fold. At normal Ca(2+) concentrations, motility increased by 290% when the extracellular K(+) concentration was reduced 10-fold (from control of 5.4 mM to 0.54 mM), and increased to 250% of control levels when the K(+) concentration was increased between 30 and 54 mM, but was relatively uninfluenced at K(+) concentrations between 5 and 30 mM. Simultaneous application of low concentrations (20 microM) of GdCl(3) completely prevented the effects of low and high K(+) on motility. Exposure to Gd(3+) or Tb(3+) also produced a flattening of the cells and enhanced cell attachment. Although the steady state intracellular Ca(2+) concentration was not significantly influenced by the K(+) concentration, the resting permeability to divalent cations, determined from Mn(2+) quench rates in fura-loaded cells, was significantly increased by a reduction in the K(+) concentration. These results indicate that resting Ca(2+) influx is critical to the movement of B16F10 melanoma cells, and demonstrate that lanthanides, which block resting Ca(2+) influx pathways, are potent antimotility agents.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Terbium(III) chloride hexahydrate, 99.9% trace metals basis
Sigma-Aldrich
Terbium(III) chloride hexahydrate, 99.999% trace metals basis
Sigma-Aldrich
Terbium(III) chloride, anhydrous, powder, 99.9% trace metals basis
Sigma-Aldrich
Terbium(III) chloride, anhydrous, powder, 99.99% trace metals basis