Potential Role of Sodium-Proton Exchangers in the Low Concentration Arsenic Trioxide-Increased Intracellular pH and Cell Proliferation
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Date
2012
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PUBLIC LIBRARY SCIENCE
Abstract
Arsenic main inorganic compound is arsenic trioxide (ATO) presented in solution mainly as arsenite. ATO increases intracellular pH (pHi), cell proliferation and tumor growth. Sodium-proton exchangers (NHEs) modulate the pHi, with NHE1 playing significant roles. Whether ATO-increased cell proliferation results from altered NHEs expression and activity is unknown. We hypothesize that ATO increases cell proliferation by altering pHi due to increased NHEs-like transport activity. Madin-Darby canine kidney (MDCK) cells grown in 5 mmol/L D-glucose-containing DMEM were exposed to ATO (0.05, 0.5 or 5 mu mol/L, 0-48 hours) in the absence or presence of 5-N, N-hexamethylene amiloride (HMA, 5-100 mu mol/L, NHEs inhibitor), PD-98059 (30 mu mol/L, MAPK1/2 inhibitor), Go6976 (10 mu mol/L, PKC alpha, beta I and mu inhibitor), or Schering 28080 (10 mu mol/L, H+/K(+)ATPase inhibitor) plus concanamycin (0.1 mu mol/L, V type ATPases inhibitor). Incorporation of [H-3]thymidine was used to estimate cell proliferation, and counting cells with a hemocytometer to determine the cell number. The pHi was measured by fluorometry in 2,7-bicarboxyethyl-5,6-carboxyfluorescein loaded cells. The Na+-dependent HMA-sensitive NHEs-like mediated proton transport kinetics, NHE1 protein abundance in the total, cytoplasm and plasma membrane protein fractions, and phosphorylated and total p42/44 mitogen-activated protein kinases (p42/44(mapk)) were also determined. Lowest ATO (0.05 mu mol/L, similar to 0.01 ppm) used in this study increased cell proliferation, pHi, NHEs-like transport and plasma membrane NHE1 protein abundance, effects blocked by HMA, PD-98059 or Go6976. Cell-buffering capacity did not change by ATO. The results show that a low ATO concentration increases MDCK cells proliferation by NHEs (probably NHE1)-like transport dependent-increased pHi requiring p42/44(mapk) and PKC alpha, beta I and/or mu activity. This finding could be crucial in diseases where uncontrolled cell growth occurs, such as tumor growth, and in circumstances where ATO, likely arsenite, is available at the drinking-water at these levels. Citation: Aravena C, Beltran AR, Cornejo M, Torres V, Diaz ES, et al. (2012) Potential Role of Sodium-Proton Exchangers in the Low Concentration Arsenic Trioxide-Increased Intracellular pH and Cell Proliferation. PLoS ONE 7(12): e51451. doi:10.1371/journal.pone.0051451
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Keywords
EPITHELIAL DOME FORMATION, VEIN ENDOTHELIAL-CELLS, PROTEIN-KINASE-C, NA+/H+ EXCHANGER, CANCER CELLS, NHE3 EXPRESSION, DRINKING-WATER, SMOOTH-MUSCLE, DNA-SYNTHESIS, NITRIC-OXIDE