We explored possibility that sodium/calcium exchanger 1 (NCX1) is involved in pH modulation and apoptosis induction in GYY4137 treated cells. We have shown that although 10 days treatment with GYY4137 did not significantly decreased volume of tumors induced by colorectal cancer DLD1 cells in nude mice, it already induced apoptosis in these tumors. Treatment of DLD1 and ovarian cancer A2780 cells with GYY4137 resulted in intracellular acidification in a concentration-dependent manner. We observed increased mRNA and protein expression of both, NCX1 and sodium/hydrogen exchanger 1 (NHE1) in DLD1-induced tumors from GYY4137-treated mice. NCX1 was coupled with NHE1 in A2780 and DLD1 cells and this complex partially disintegrated after GYY4137 treatment. We proposed that intracellular acidification is due to uncoupling of NCX1/NHE1 complex rather than blocking of the reverse mode of NCX1, probably due to internalization of NHE1. Results might contribute to understanding molecular mechanism of H2S-induced apoptosis in tumor cells.
- Keywords
- Apoptosis, Hydrogen sulfide, Intracellular acidification, Sodium/calcium exchanger, Sodium/hydrogen exchanger,
- MeSH
- Apoptosis drug effects MeSH
- Hydrogen-Ion Concentration MeSH
- Humans MeSH
- Morpholines pharmacology MeSH
- Mice, Nude MeSH
- Cell Line, Tumor MeSH
- Organothiophosphorus Compounds pharmacology MeSH
- Cell Proliferation drug effects MeSH
- Antineoplastic Agents pharmacology MeSH
- Sodium-Calcium Exchanger metabolism MeSH
- Sodium-Hydrogen Exchanger 1 metabolism MeSH
- Hydrogen Sulfide metabolism MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- GYY 4137 MeSH Browser
- Morpholines MeSH
- NCX1 protein, mouse MeSH Browser
- Organothiophosphorus Compounds MeSH
- Antineoplastic Agents MeSH
- Sodium-Calcium Exchanger MeSH
- SLC9A1 protein, human MeSH Browser
- Slc9a1 protein, mouse MeSH Browser
- Sodium-Hydrogen Exchanger 1 MeSH
- sodium-calcium exchanger 1 MeSH Browser
- Hydrogen Sulfide MeSH
Avian leukosis virus subgroup J (ALV-J) is an important concern for the poultry industry. Replication of ALV-J depends on a functional cellular receptor, the chicken Na+/H+ exchanger type 1 (chNHE1). Tryptophan residue number 38 of chNHE1 (W38) in the extracellular portion of this molecule is a critical amino acid for virus entry. We describe a CRISPR/Cas9-mediated deletion of W38 in chicken primordial germ cells and the successful production of the gene-edited birds. The resistance to ALV-J was examined both in vitro and in vivo, and the ΔW38 homozygous chickens tested ALV-J-resistant, in contrast to ΔW38 heterozygotes and wild-type birds, which were ALV-J-susceptible. Deletion of W38 did not manifest any visible side effect. Our data clearly demonstrate the antiviral resistance conferred by precise CRISPR/Cas9 gene editing in the chicken. Furthermore, our highly efficient CRISPR/Cas9 gene editing in primordial germ cells represents a substantial addition to genotechnology in the chicken, an important food source and research model.
- Keywords
- CRISPR/Cas9 genome editing in chicken, Na+/H+ exchanger type 1, avian leukosis virus subgroup J, disease resilience in poultry, primordial germ cells,
- MeSH
- CRISPR-Cas Systems MeSH
- Gene Editing MeSH
- Animals, Genetically Modified genetics immunology virology MeSH
- Chickens MeSH
- Poultry Diseases genetics immunology virology MeSH
- Disease Resistance MeSH
- Avian Leukosis genetics immunology virology MeSH
- Avian Proteins genetics immunology MeSH
- Sodium-Hydrogen Exchanger 1 genetics immunology MeSH
- Avian Leukosis Virus classification genetics physiology MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Avian Proteins MeSH
- Sodium-Hydrogen Exchanger 1 MeSH
BACKGROUND: Na(+)/H(+) exchanger-1 (NHE-1) is involved in pH regulation and is up-regulated in different malignancies. Activation of NHE-1 is one way for allowing cells to avoid intracellular acidification and protect them against apoptosis. Inhibitors of NHE-1 are able to decrease intracellular pH and induce apoptosis. Some statins can also act by partial inhibition of NHE-1. This review presents progress in understanding the mechanisms of action of these inhibitors, connections with certain genetic mutations and acquired treatment resistance, as well as new patents on them. METHODS: A MEDLINE search for original and review articles using key terms, Na(+)/H(+) exchanger, leukemia, cariporide, and amiloride. Recent patents with NHE-1 inhibitors published by United States Patent and Trademark Office are also presented. RESULTS AND CONCLUSIONS: Sorafenib is used for the treatment of acute myeloid leukemia patients carrying internal tandem duplication of fms-like tyrosine kinase 3 (FLT3-ITD) mutation. 5-(N, N-hexamethylene)-amiloride can increase the suppression of FLT3 signaling by sorafenib. NHE-1 inhibitors are able to increase the sensitivity of chronic myeloid leukemia cells to tyrosine kinase inhibitors, including through the inhibition of P-glycoprotein. NHE-1 inhibitors are promising adjuvant drugs for overcoming acquired resistance to treatment in various malignant hemopathies.
- Keywords
- BCR/ABL, FLT3/ITD, Na+/H+ exchanger, P-glycoprotein, amiloride, apoptosis, cariporide, heme oxygenase-1, imatinib mesylate, intracellular pH, isoprenylation, leukemia, lovastatin, sorafenib, statins,
- MeSH
- Leukemia, Myeloid, Acute drug therapy genetics MeSH
- Amiloride pharmacology MeSH
- Apoptosis drug effects MeSH
- Leukemia, Myelogenous, Chronic, BCR-ABL Positive drug therapy genetics MeSH
- Phenylurea Compounds pharmacology MeSH
- Genes, abl genetics MeSH
- Guanidines pharmacology MeSH
- Heme Oxygenase-1 antagonists & inhibitors metabolism MeSH
- Imatinib Mesylate pharmacology MeSH
- Protein Kinase Inhibitors pharmacology MeSH
- Hydrogen-Ion Concentration MeSH
- Drug Interactions MeSH
- Humans MeSH
- Mutation genetics MeSH
- Sodium-Hydrogen Exchangers antagonists & inhibitors physiology MeSH
- Tumor Hypoxia physiology MeSH
- Cell Line, Tumor MeSH
- Niacinamide analogs & derivatives pharmacology MeSH
- Osmolar Concentration MeSH
- Patents as Topic MeSH
- DNA Damage physiology MeSH
- Cation Transport Proteins antagonists & inhibitors physiology MeSH
- Antineoplastic Agents pharmacology MeSH
- Signal Transduction drug effects MeSH
- Sodium-Hydrogen Exchanger 1 MeSH
- Sorafenib MeSH
- Hydroxymethylglutaryl-CoA Reductase Inhibitors pharmacokinetics MeSH
- Sulfones pharmacology MeSH
- fms-Like Tyrosine Kinase 3 antagonists & inhibitors genetics MeSH
- Up-Regulation physiology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Review MeSH
- Names of Substances
- Amiloride MeSH
- cariporide MeSH Browser
- Phenylurea Compounds MeSH
- FLT3 protein, human MeSH Browser
- Guanidines MeSH
- Heme Oxygenase-1 MeSH
- Imatinib Mesylate MeSH
- Protein Kinase Inhibitors MeSH
- Sodium-Hydrogen Exchangers MeSH
- Niacinamide MeSH
- Cation Transport Proteins MeSH
- Antineoplastic Agents MeSH
- SLC9A1 protein, human MeSH Browser
- Sodium-Hydrogen Exchanger 1 MeSH
- Sorafenib MeSH
- Hydroxymethylglutaryl-CoA Reductase Inhibitors MeSH
- Sulfones MeSH
- fms-Like Tyrosine Kinase 3 MeSH
Saccharomyces cerevisiae, Schizosaccharomyces pombe, Endomyces magnussi, Lodderomyces elongisporus and Rhodotorula gracilis, yeast species ranging from a glycolytic type to a strictly aerobic one, were tested for the activity of their plasma membrane H(+)-ATPase and the effect of alkaline metal cations thereon. The ATP-hydrolyzing activity of membranes from glucose-activated cells ranged from 456 to 932 mumol inorganic phosphate released per min per 1 g membrane protein. The effect of 0.2 M Li+, Na+, K+, Rb+ and Cs+ never exceeded the statistical range of error. In contrast, acidification after glucose addition ranged from 0.15 (for R. gracilis) to 14.8 nmol H+ per min per mg dry weight (for S. cerevisiae) and it was markedly influenced by the presence of alkaline metal chlorides, the highest effect observed being a seven-fold increase by K+ in a S. cerevisiae suspension. The effects were additive to those observed without ions in solution and are ascribed to the operation of independent channels and/or exchange systems for H+ with a clear selectivity toward K+. The separate nature of the ion-triggered extracellular acidification is supported by a different ratio of titration to pH-derived acidity with and without K+.
- MeSH
- Adenosine Triphosphate metabolism MeSH
- Cell Membrane enzymology MeSH
- Hydrolysis MeSH
- Cations, Monovalent metabolism MeSH
- Hydrogen-Ion Concentration MeSH
- Metals, Alkaline Earth metabolism MeSH
- Yeasts enzymology MeSH
- Proton-Translocating ATPases metabolism MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Adenosine Triphosphate MeSH
- Cations, Monovalent MeSH
- Metals, Alkaline Earth MeSH
- Proton-Translocating ATPases MeSH
Subgroup J avian leukosis virus (ALV-J) is unique among the avian sarcoma and leukosis viruses in using the multimembrane-spanning cell surface protein Na(+)/H(+) exchanger type 1 (NHE1) as a receptor. The precise localization of amino acids critical for NHE1 receptor activity is key in understanding the virus-receptor interaction and potential interference with virus entry. Because no resistant chicken lines have been described until now, we compared the NHE1 amino acid sequences from permissive and resistant galliform species. In all resistant species, the deletion or substitution of W38 within the first extracellular loop was observed either alone or in the presence of other incidental amino acid changes. Using the ectopic expression of wild-type or mutated chicken NHE1 in resistant cells and infection with a reporter recombinant retrovirus of subgroup J specificity, we studied the effect of individual mutations on the NHE1 receptor capacity. We suggest that the absence of W38 abrogates binding of the subgroup J envelope glycoprotein to ALV-J-resistant cells. Altogether, we describe the functional importance of W38 for virus entry and conclude that natural polymorphisms in NHE1 can be a source of host resistance to ALV-J.
- MeSH
- Virus Internalization * MeSH
- DNA Mutational Analysis MeSH
- Sodium-Hydrogen Exchangers genetics metabolism MeSH
- Birds MeSH
- Viral Tropism * MeSH
- Tryptophan genetics metabolism MeSH
- Receptors, Virus genetics metabolism MeSH
- Avian Leukosis Virus physiology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Sodium-Hydrogen Exchangers MeSH
- Tryptophan MeSH
- Receptors, Virus MeSH
AtChx17p is a putative K(+)/H(+) exchanger from Arabidopsis thaliana, expressed in the roots and probably involved in K(+) acquisition and homeostasis. AtCHX17 cDNA complements the phenotypes of the kha1Delta mutation in S. cerevisiae cells: a growth defect at increased pH and hygromycin sensitivity. The localization of GFP-tagged AtChx17 protein in yeast cells is similar to that of ScKha1p: a bold dotted pattern inside the cells resembling the Golgi fluorescence markers. These results show that (a) the proteins AtChx17 and ScKha1 could have similar functions and (b) S. cerevisiae kha1 deletion mutants could serve for the heterologous expression and characterization of plant transporters. The results of this work are evidence that a S. cerevisiae strain with deletions of genes encoding alkali-metal-cation/H(+) antiporters (i.e. Nha1p, Nhx1p, Kha1p) could be an ideal tool for expression and functional analysis of any type of similar plant antiporters (plasma membrane, endosomal/prevacuolar and Golgi).
- MeSH
- Arabidopsis genetics MeSH
- Cinnamates pharmacology MeSH
- Gene Deletion MeSH
- Potassium-Hydrogen Antiporters genetics MeSH
- Phenotype MeSH
- Hygromycin B analogs & derivatives pharmacology MeSH
- DNA, Complementary MeSH
- Hydrogen-Ion Concentration MeSH
- Sodium-Hydrogen Exchangers genetics MeSH
- Arabidopsis Proteins genetics MeSH
- Saccharomyces cerevisiae Proteins genetics MeSH
- Saccharomyces cerevisiae drug effects genetics growth & development MeSH
- Genetic Complementation Test MeSH
- Green Fluorescent Proteins genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- CHX17 protein, Arabidopsis MeSH Browser
- Cinnamates MeSH
- Potassium-Hydrogen Antiporters MeSH
- hygromycin A MeSH Browser
- Hygromycin B MeSH
- KHA1 protein, S cerevisiae MeSH Browser
- DNA, Complementary MeSH
- Sodium-Hydrogen Exchangers MeSH
- Arabidopsis Proteins MeSH
- Saccharomyces cerevisiae Proteins MeSH
- Green Fluorescent Proteins MeSH
BACKGROUND/AIMS: Melatonin is a hormone transferring information about duration of darkness to the organism and is known to modulate several signaling pathways in the cells, e.g. generation of endoplasmic reticulum stress, oxidative status of the cells, etc. Melatonin has been shown to exert antiproliferative and cytotoxic effects on various human cancers. We proposed that this hormone can differently affect tumour cells and healthy cells. METHODS: We compared the effect of 24 h melatonin treatment on calcium transport (by fluorescent probes FLUO-3AM and Rhod-5N), ER stress (determined as changes in the expression of CHOP, XBP1 and fluorescently, using Thioflavin T), ROS formation (by CellROX® Green/Orange Reagent) and apoptosis induction (by Annexin-V-FLUOS/propidiumiodide) in two tumour cell lines - ovarian cancer cell line A2780 and stable cell line DLD1 derived from colorectal carcinoma, with non-tumour endothelial cell line EA.hy926. RESULTS: Melatonin increased apoptosis in both tumour cell lines more than twice, while in EA.hy926 cells the apoptosis was increased only by 30%. As determined by silencing with appropriate siRNAs, both, type 1 sodium/calcium exchanger and type 1 IP3 receptor are involved in the apoptosis induction. Antioxidant properties of melatonin were significantly increased in EA.hy926 cells, while in tumour cell lines this effect was much weaker. CONCLUSION: Taken together, melatonin has different antioxidative effects on tumour cells compared to non-tumour ones; it also differs in the ability to induce apoptosis through the type 1 sodium/calcium exchanger, and type 1 IP3 receptor. Different targeting of calcium transport systems in tumour and normal, non-tumour cells is suggested as a key mechanism how melatonin can exert its anticancer effects. Therefore, it might have a potential as a novel therapeutic implication in cancer treatment.
- Keywords
- Apoptosis, Calcium, Cancer, ER-stress, Melatonin, Reactive oxygen species,
- MeSH
- Apoptosis drug effects MeSH
- Cytosol metabolism MeSH
- Microscopy, Fluorescence MeSH
- Inositol 1,4,5-Trisphosphate Receptors antagonists & inhibitors genetics MeSH
- Humans MeSH
- RNA, Small Interfering metabolism MeSH
- Melatonin toxicity MeSH
- Cell Line, Tumor MeSH
- Sodium-Calcium Exchanger antagonists & inhibitors genetics metabolism MeSH
- Reactive Oxygen Species metabolism MeSH
- RNA Interference MeSH
- Endoplasmic Reticulum Stress drug effects MeSH
- Transcription Factor CHOP genetics metabolism MeSH
- Calcium metabolism MeSH
- X-Box Binding Protein 1 genetics metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Inositol 1,4,5-Trisphosphate Receptors MeSH
- RNA, Small Interfering MeSH
- Melatonin MeSH
- Sodium-Calcium Exchanger MeSH
- Reactive Oxygen Species MeSH
- sodium-calcium exchanger 1 MeSH Browser
- Transcription Factor CHOP MeSH
- Calcium MeSH
- Xbp1 protein, mouse MeSH Browser
- X-Box Binding Protein 1 MeSH
- MeSH
- Gene Frequency MeSH
- Genetic Linkage * MeSH
- Chromosome Mapping veterinary MeSH
- Swine genetics MeSH
- Proton-Translocating ATPases genetics MeSH
- Sodium-Potassium-Exchanging ATPase genetics MeSH
- Vacuolar Proton-Translocating ATPases * MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Proton-Translocating ATPases MeSH
- Sodium-Potassium-Exchanging ATPase MeSH
- Vacuolar Proton-Translocating ATPases * MeSH
- MeSH
- Gene Frequency MeSH
- Genetic Linkage * MeSH
- Radiation Hybrid Mapping MeSH
- Molecular Sequence Data MeSH
- Swine genetics MeSH
- Protein Precursors genetics MeSH
- Zebrafish Proteins * MeSH
- Sodium-Potassium-Exchanging ATPase genetics MeSH
- Vacuolar Proton-Translocating ATPases genetics MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- atp1a1b protein, zebrafish MeSH Browser
- involucrin MeSH Browser
- Protein Precursors MeSH
- Zebrafish Proteins * MeSH
- Sodium-Potassium-Exchanging ATPase MeSH
- Vacuolar Proton-Translocating ATPases MeSH
BACKGROUND: Disorders in sodium metabolism such as an increased total body exchangeable sodium, were found in diabetic patients, although the underlying mechanisms were not clear. The aim of the study was to evaluate red blood cell sodium transport in patients with insulin dependent diabetes mellitus (IDDM) without diabetic nephropathy. METHODS AND RESULTS: Renal hemodynamics using the clearance of inulin and para-amino-hippuric acid during euglycemic clamp and red blood cell sodium transport were examined in 13 IDDM patients without microalbuminuria and in 12 weight-, age- and sex-matched healthy controls. Despite normal renal hemodynamics and intracellular sodium concentrations (6.57 +/- 1.45 vs 5.95 +/- 0.60 mmol/l), IDDM patients had lowered clearance of sodium (2.22 +/_ 1.11 vs 3.24 +/- 1.32 ml/min; p < 0.01) and increased activity of natrium-lithium countertransport compared to C (0.76 +/- 0.50 vs 0.31 +/- 0.22 mmol.l-1 .h-1; p < 0.01). No significant differences between IDDM and C were found in Na+-K+ pump (7.95 +/- 1.95 vs 6.9 +/- 0.99 mmol.l-1 .h-1), in Na+-K+ cotransport (0.68 +/- 0.82 vs 0.82 +/- 0.71 mmol.l-1 .h-1) and in passive Na+ permeability (0.11 +/- 0.05 vs 0.09 +/- 0.02 mmoll.l-1 .h-1). CONCLUSIONS: IDDM patients without signs of diabetic nephropathy have shown changes in sodium-lithium countertransport which could play a role in the pathogenesis of diabetic nephropathy and hypertension in the course of the disease.
- MeSH
- Biological Transport MeSH
- Diabetes Mellitus, Type 1 blood MeSH
- Adult MeSH
- Erythrocyte Membrane metabolism MeSH
- Humans MeSH
- Sodium blood MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Publication type
- English Abstract MeSH
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Sodium MeSH
