pH-Channeling in Cancer: How pH-Dependence of Cation Channels Shapes Cancer Pathophysiology.

Tissue acidosis plays a pivotal role in tumor progression: in particular, interstitial acidosis promotes tumor cell invasion, and is a major contributor to the dysregulation of tumor immunity and tumor stromal cells. The cell membrane and integral membrane proteins commonly act as important sensors and transducers of altered pH. Cell adhesion molecules and cation channels are prominent membrane proteins, the majority of which is regulated by protons. The pathophysiological consequences of proton-sensitive ion channel function in cancer, however, are scarcely considered in the literature. Thus, the main focus of this review is to highlight possible events in tumor progression and tumor immunity where the pH sensitivity of cation channels could be of great importance

2-Aminophenoxazine-3-one and 2-amino-4,4α-dihydro-4α,7-dimethyl-3H-phenoxazine-3-one cause cellular apoptosis by reducing higher intracellular pH in cancer cells

We examined intracellular pH (pHi) of ten cancer cell lines derived from different organs and two normal cell lines including human embryonic lung fibroblast cells (HEL) and human umbilical vein endothelial cells (HUVEC) in vitro, and found that pHi of most of these cancer cells was evidently higher (pH 7.5 to 7.7) than that of normal cells (7.32 and 7.44 for HEL and HUVEC, respectively) and that of primary leukemic cells and erythrocytes hitherto reported (≤7.2). Higher pHi in these cancer cells could be related to the Warburg effect in cancer cells with enhanced glycolytic metabolism. Since reversal of the Warburg effect may perturb intracellular homeostasis in cancer cells, we looked for compounds that cause extensive reduction of pHi, a major regulator of the glycolytic pathway and its associated metabolic pathway. We found that phenoxazine compounds, 2-aminophenoxazine-3-one (Phx-3) and 2-amino-4,4α-dihydro-4α,7-dimethyl-3H-phenoxazine-3-one (Phx-1) caused a rapid and drastic dose-dependent decrease of pHi in ten different cancer cells within 30 min, though the extent of the decrease of pHi was significantly larger for Phx-3 (ΔpHi = 0.6 pH units or more for 100 µM Phx-3) than for Phx-1 (ΔpHi = 0.1 pH units or more for 100 µM Phx-1). This rapid and drastic decrease of pHi in a variety of cancer cells caused by Phx-3 and Phx-1 possibly perturbed their intracellular homeostasis, and extensively affected the subsequent cell death, because these phenoxazines exerted dose-dependent proapoptotic and cytotoxic effects on these cells during 72 h incubation, confirming a causal relationship between ΔpHi and cytotoxic effects due to Phx-3 and Phx-1. Phx-3 and Phx-1 also reduced pHi of normal cells including HEL and HUVEC, although they exerted less proapoptotic and cytotoxic effects on these cells than on cancer cells. Drugs such as Phx-3 and Phx-1 that reduce pHi and thereby induce cellular apoptosis might serve as benevolent anticancer drugs

Intestinal glycyl-L-phenylalanine and L-phenylalanine transport in a euryhaline teleost

The transport mechanisms for the dipeptide glycyl-L-phenylalanine (Gly-Phe) and L-phenylalanine (Phe) were characterized in fish intestinal brush-border membrane vesicles (BBMV). Gly-Phe was rapidly hydrolyzed only intravesicularly with almost total hydrolysis occurring even at 10 s. Dipeptide uptake was not stimulated by an inward gradient of Na, K, or H. Phe uptake was stimulated by an inward gradient of either Na or K but displayed an overshoot phenomenon only in the presence of an Na gradient. Kinetic analysis of the effect of substrate concentration on transport rate revealed that transport of both Gly-Phe and Phe occurred by a saturable process conforming to Michaelis-Menten kinetics. The K(m) for Gly-Phe was 9.8 ± 3.5 mM, whereas that for Phe in the presence of Na or K, respectively, was 0.74 ± 0.13 and 1.1 ± 0.37 mM. Maximum uptake for Gly-Phe and for Phe in the presence of Na and K was 5.1, 0.9, and 0.4 nmol·mg and protein-1·5 s-1, respectively. Gly-Phe and Phe transport displayed different patterns of inhibition by dipeptides and amino acids. These results suggest that Gly-Phe and Phe are transported via different mechanisms, with Gly-Phe being hydrolyzed during a carrier-mediated, cation-independent process and Phe being transferred via a Na+ cotransport process similar to that described in mammals. During conditions of high luminal dipeptide concentrations, the Gly-Phe pathway may make a significant contribution to total Phe uptake

Intestinal glucose transport and salinity adaptation in a euryhaline teleost

Glucose transport by upper and lower intestinal brush-border membrane vesicles of the African tilapia (Oreochromis mossambicus) was characterized in fish acclimated to either freshwater or full-strength seawater. D-[3H]-glucose uptake by vesicles was stimulated by a transmembrane Na gradient, was electrogenic, and was enhanced by counter-transport of either D-glucose or D-galactose. Glucose transport was greater in the upper intestine than in the lower intestine and in seawater animals rather than in fish acclimated to freshwater. Glucose influx (10-s uptake) involved both saturable and nonsaturable transport components. Seawater adaptation increased apparent glucose influx K(t), J(max), apparent diffusional permeability (P), and the apparent Na affinity of the cotransport system in both intestinal segments, but the stoichiometry of Na-glucose transfer (1:1) was unaffected by differential saline conditions or gut region. It is suggested that increased sugar transport in seawater animals is due to the combination of enhanced Na-binding properties and an increase in number of transfer rate of the transport proteins. Freshwater animals compensate for reduced Na affinity of the coupled process by markedly increasing the protein affinity for glucose

Brush-border inositol transport by intestines of carnivorous and herbivorous teleosts

Transport characteristics of myoinositol by isolated brush-border membrane vesicles of two fish, the herbivorous tilapia (Oreochromis mossambicus) and the carnivorous eel (Anguilla anguilla), were measured. [3H]myoinositol uptake by vesicles of both fish was stimulated by a transmembrane Na gradient, was electrogenic, and was inhibited by phloridzin. Kinetic analysis of myoinositol influx disclosed species differences (tilapia, K = 0.15 mM, J(max) = 0.2 nmol·mg protein-1·min-1; eel, K = 2.6 mM, J(max) = 0.8 nmol·mg protein-1·min-1). D-Glucose inhibition of myoinositol influx was shown to be noncompetitive. Additional inhibition studies with a range of sugars demonstrated that aldohexoses in the C-1 chair conformation were preferred substrates. Myoinositol had no effect on D-glucose transport. Preloading vesicles with myoinositol transstimulated [3H]myoinositol uptake, while the use of internal D-glucose was without effect. These results suggest that the intestinal brush border may have a pathway for myoinositol transport entirely separate from that for D-glucose but inhibited by D-glucose via binding to a regulator site on the myoinositol transporter. Markedly dissimilar influx kinetic constants suggest possible differences in myoinositol needs by carnivorous and herbivorous fish

Dietary hormonal modification of growth intestinal ATPase, and glucose transport in tilapia

The effect of growth stimulatory and inhibitory dietary applications of hormones [3,5,3'-triiodo-L-thyronine (T3) and 17α-methyltestosterone (MT)] on Na+-K+-adenosinetriphosphatase (ATPase) activity and glucose transport by upper and lower intestinal brush-border membrane vesicles of tilapia (Oreochromis mossambicus) were characterized. Both enzyme activity and glucose transport were greater in growth-stimulatory treatments and lower in growth-inhibitory treatments than in the control. Growth on stimulatory hormone treatments increased apparent glucose influx kinetics (one-half maximum glucose influx, maximum glucose influx, and apparent diffusion coefficient) in both intestinal segments, whereas inhibitory treatments reduced these parameters in upper intestine but had no effect on these parameters in lower intestine. All hormone treatments increased the stoichiometry of Na-glucose cotransport from 1:1 in the control to 2:1 under test conditions. It is suggested that observed patterns of altered growth are due, in part, to hormonally modified intestinal nutrient transport and Na+-K+-ATPase activities

Effects of beta-endorphin and Naloxone on in vitro maturation of bovine oocytes

Bovine cumulus-oocyte complexes (COCs) and mural granulosa cells express the mRNA coding for the p-opioid receptor. The addition of beta-enclorphin (beta-end) to oocytes cultured in hormonally-supplemented in vitro maturation (IVM) medium had no effect on the rates of oocytes reaching the Metaphase II (MII) stage, but significantly decreased the maturation rate (P < 0.05) and arrested oocytes at metaphase I (MI) after culture in hormone-free medium (P < 0.001). Naloxone (Nx) reverted this inhibitory effect of beta-end. Moreover, Nx "per se" showed a dose-dependent dual effect. When added at high concentration (10(-3) M), it significantly reduced the rate of oocytes in MII (P < 0.001), thus increasing the rate of oocytes arrested in MI. However, Nx added at low concentration (10(-8) M) significantly increased oocyte maturation (P < 0.001). High concentration of Nx induced an increase in both intracellular calcium concentration ([Ca2+](i)) and in the activity of the mitogen-activated protein kinase (MAPK) also called extracellular-regulated kinase (ERK) in cumulus cells of bovine COCs. Blocking the rise in [Ca2+](i) with the calcium chelator acetoxymethylester-derived form of bis (o-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM) reversed the Nx-dependent inhibition of meiotic maturation observed at high Nx concentrations. Whereas blocking ERK with the MAPK/ERK kinase (MEK) inhibitor, PD98059, had no effect on this process. Therefore, we concluded that the p-opioid receptor, by inducing [Ca2+](i) increase, participates in the cumulus-oocyte coupled signaling associated with oocyte maturation. (C) 2002 Wiley-Liss, Inc