• MeSH
• koronární nemoc farmakoterapie   MeSH
• Na(+)-H(+) antiport antagonisté a inhibitory   MeSH
• Publikační typ
• kongresy MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• kardiologie
• angiologie

• MeSH
• členovci MeSH
• iontová výměna MeSH
• sodík analýza   MeSH
• vápník analýza   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

• MeSH
• iontová výměna MeSH
• krysa rodu rattus MeSH
• membrány MeSH
• svaly MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

• MeSH
• buněčná membrána MeSH
• cytologické techniky MeSH
• iontová výměna MeSH
• ryby MeSH
• sarkolema MeSH

The sodium/calcium exchanger (NCX) type 1 has been well described in various cancers, but little is known about the other two NCX types (NCX2 and NCX3). In this study, we used the selective blocker of NCX3 - YM-244769 to investigate changes in apoptosis induction, migration, proliferation, intracellular calcium and ATP in four cancer cell lines - DLD1, HeLa, MDA-MB-231 and JIMT1. In all four cell lines we observed a concentration-dependent increase in the number of apoptotic cells, as well as reduced migration and proliferation. Induction of hypoxic conditions did not alter the response of these cells to YM-244769 in any of the above-mentioned parameters. These results indicate the role of NCX3 in cancer cell migration, proliferation and apoptosis, as inhibition of NCX1 by the specific blocker SEA0400 had no significant effect on these parameters. However, we verified the effect of NCX3 inhibition by using CRISPR/Cas9 to generate clones in which the SLC8A3 (NCX3) gene was deleted, and we obtained the same results. In addition, mitochondrial respiration was impaired in the clones with NCX3 knocked-out, suggesting that NCX3 also play a role in bioenergetics. In conclusion, we have clearly shown that NCX3 plays an important anti-apoptotic, pro-migratory and proliferative role in the cancer cells by affecting mitochondrial bioenergetics, thus supporting their survival and fate.

• MeSH
• apoptóza účinky léků   MeSH
• lidé MeSH
• mitochondrie metabolismus   účinky léků   MeSH
• nádorové buněčné linie MeSH
• nádory * metabolismus   patologie   genetika   MeSH
• pohyb buněk účinky léků   MeSH
• proliferace buněk účinky léků   MeSH
• pumpa pro výměnu sodíku a vápníku * metabolismus   genetika   antagonisté a inhibitory   MeSH
• vápník metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• chromatografie iontoměničová MeSH
• kovy analýza   MeSH
• léčivé přípravky analýza   MeSH
• roztoky analýza   MeSH
• Publikační typ
• farmakopea MeSH

Given the potential clinical benefit of inhibiting Na+/Ca2+ exchanger (NCX) activity during myocardial ischemia reperfusion (I/R), pharmacological approaches have been pursued to both inhibit and clarify the importance of this exchanger. SEA0400 was reported to have a potent NCX selectivity. Thus, we examined the effect of SEA0400 on NCX currents and I/R induced intracellular Ca2+ overload in mouse ventricular myocytes using patch clamp techniques and fluorescence measurements. Ischemia significantly inhibited inward and outward NCX current (from -0.04±0.01nA to 0 nA at -100 mV; from 0.23±0.08 nA to 0.11±0.03 nA at +50 mV, n=7), Subsequent reperfusion not only restored the current rapidly but enhanced the current amplitude obviously, especially the outward currents (from 0.23±0.08 nA to 0.49±0.12 nA at +50 mV, n=7). [Ca2+]i, expressed as the ratio of Fura-2 fluorescence intensity, increased to 138±7 % (P<0.01) during ischemia and to 210±11 % (P<0.01) after reperfusion. The change of NCX current and the increase of [Ca2+]i during I/R can be blocked by SEA0400 in a dose-dependent manner with an EC50 value of 31 nM and 28 nM for the inward and outward NCX current, respectively. The results suggested that SEA0400 is a potent NCX inhibitor, which can protect mouse cardiac myocytes from Ca2+ overload during I/R injuries.

• MeSH
• aniliny terapeutické užití   MeSH
• fenylethery terapeutické užití   MeSH
• financování vládou MeSH
• ischemická choroba srdeční etiologie   komplikace   MeSH
• kardiomyocyty metabolismus   MeSH
• metoda terčíkového zámku metody   využití   MeSH
• myši inbrední C57BL MeSH
• pumpa pro výměnu sodíku a vápníku antagonisté a inhibitory   MeSH
• reperfuze myokardu MeSH
• Publikační typ
• srovnávací studie MeSH

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.

• MeSH
• apoptóza účinky léků   MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• morfoliny farmakologie   MeSH
• myši nahé MeSH
• nádorové buněčné linie MeSH
• organothiofosforové sloučeniny farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• protinádorové látky farmakologie   MeSH
• pumpa pro výměnu sodíku a vápníku metabolismus   MeSH
• sodíko-vodíkový výměnný transportér 1 metabolismus   MeSH
• sulfan metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Na(+)/Ca(2+) exchanger (NCX) proteins operate through the alternating access mechanism, where the ion-binding pocket is exposed in succession either to the extracellular or the intracellular face of the membrane. The archaeal NCX_Mj (Methanococcus jannaschii NCX) system was used to resolve the backbone dynamics in the inward-facing (IF) and outward-facing (OF) states by analyzing purified preparations of apo- and ion-bound forms of NCX_Mj-WT and its mutant, NCX_Mj-5L6-8. First, the exposure of extracellular and cytosolic vestibules to the bulk phase was evaluated as the reactivity of single cysteine mutants to a fluorescent probe, verifying that NCX_Mj-WT and NCX_Mj-5L6-8 preferentially adopt the OF and IF states, respectively. Next, hydrogen-deuterium exchange-mass spectrometry (HDX-MS) was employed to analyze the backbone dynamics profiles in proteins, preferentially adopting the OF (WT) and IF (5L6-8) states either in the presence or absence of ions. Characteristic differences in the backbone dynamics were identified between apo NCX_Mj-WT and NCX_Mj-5L6-8, thereby underscoring specific conformational patterns owned by the OF and IF states. Saturating concentrations of Na(+) or Ca(2+) specifically modify HDX patterns, revealing that the ion-bound/occluded states are much more stable (rigid) in the OF than in the IF state. Conformational differences observed in the ion-occluded OF and IF states can account for diversifying the ion-release dynamics and apparent affinity (Km ) at opposite sides of the membrane, where specific structure-dynamic elements can effectively match the rates of bidirectional ion movements at physiological ion concentrations.

• MeSH
• apoproteiny chemie   genetika   metabolismus   MeSH
• archeální proteiny chemie   genetika   metabolismus   MeSH
• buněčná membrána chemie   MeSH
• cystein chemie   MeSH
• interakční proteinové domény a motivy MeSH
• inzerční mutageneze MeSH
• kinetika MeSH
• konformace proteinů MeSH
• ligandy MeSH
• Methanocaldococcus metabolismus   MeSH
• molekulární modely * MeSH
• mutace MeSH
• peptidové fragmenty chemie   genetika   metabolismus   MeSH
• pumpa pro výměnu sodíku a vápníku chemie   genetika   metabolismus   MeSH
• rekombinantní proteiny chemie   metabolismus   MeSH
• sodík metabolismus   MeSH
• stabilita proteinů MeSH
• substituce aminokyselin MeSH
• vápník metabolismus   MeSH
• vazebná místa MeSH
• vodík-deuteriová výměna MeSH
• výpočetní biologie MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

• MeSH
• Ca2+-ATPasy biosyntéza   MeSH
• hormony štítné žlázy fyziologie   MeSH
• krysa rodu rattus MeSH
• myokard metabolismus   MeSH
• pumpa pro výměnu sodíku a vápníku biosyntéza   MeSH
• sarkolema metabolismus   MeSH
• srdce růst a vývoj   MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH