Hypertonic saline attenuates colonic tumor cell metastatic potential by activating transmembrane sodium conductance.
Hypertonic saline (HTS) suppresses tumor cell-endothelial interactions by reducing integrin expression. This translates into reduced adhesion, migration and metastatic potential. This study determined the relative contributions of hyperosmolarity and sodium-specific hypertonicity on the inhibitory effects of HTS, the intracellular pH and sodium responses to HTS and the role of cytoskeletal remodeling in these changes. Human colonic tumor cells (LS174T) were exposed to lipopolysaccharide under isotonic, hypertonic, sodium-free (N-methyl-D-glucamine), hyperosmolar (mannitol or urea), disrupted cytoskeletal (10 microg/ml cytochalasin D) conditions or in the presence of 5-(N-ethyl-N-isopropyl)amiloride (EIPA). Beta(1) integrin expression was measured flow-cytometrically. Intracellular sodium and pH were measured with confocal laser microscopic imaging. Statistical analysis was performed with analysis of variance, and P < 0.05 was considered significant. Data are represented as mean +/- SEM. Hypertonic exposure attenuated integrin expression (62.03 +/- 4.7% of control, P < 0.04). No discernible effect was observed with sodium-free or hyperosmolar solutions. HTS evoked a cellular alkalinization (by a mean 0.2 pH units) and an increase in cytosolic sodium concentration (by a mean 12.4 mM, P < 0.001) via upregulation of sodium-hydrogen exchange. Disassembly of actin microfilaments by cytochalasin D and antiporter inhibition with EIPA abrogated the effect of hypertonicity on integrin expression and intracellular sodium and pH (P < 0.05). HTS downregulates adhesion molecule expression via a hypertonic, sodium-specific, cytoskeletally mediated mechanism that involves activation of sodium-hydrogen exchange with associated changes in intracellular pH and sodium concentrations.
Shields
Conor J
CJ
Department of Surgery, National University of Ireland, Cork, Ireland. conor@narrowpoint.com
Winter
Desmond C
DC
Geibel
John P
JP
O'Sullivan
Gerald C
GC
Wang
Jiang Huai
JH
Redmond
H Paul
HP
D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D003110:Colonic Neoplasms / Q000378:metabolism*; D006801:Humans; D019012:Integrin beta1 / Q000096:biosynthesis / Q000235:genetics; D009362:Neoplasm Metastasis; D012462:Saline Solution, Hypertonic; D012964:Sodium / Q000378:metabolism*
D019012:Integrin beta1; D012462:Saline Solution, Hypertonic; D012964:Sodium