The paper is focused on pilot study of effects of novel synthetic protein kinase inhibitors-maleimide derivatives in different concentrations on normal, transformed and multipotent cell lines. Influence on cell proliferation and morphological characteristics has been demonstrated. The chosen agents cause antiproliferative effect on transformed cells and are not cytotoxic to normal cell lines. Moreover, different maleimide derivatives' effects on multipotent cells in attached and floating states has been shown. Described results can be used for further research of the maleimide derivatives as antitumor agents.
- MeSH
- buňky cytologie účinky léků MeSH
- endoteliální buňky cytologie účinky léků MeSH
- fibroblasty cytologie účinky léků MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- inhibiční koncentrace 50 MeSH
- kultivované buňky MeSH
- lidé MeSH
- maleimidy chemie farmakologie MeSH
- mezenchymální kmenové buňky cytologie účinky léků MeSH
- MFC-7 buňky MeSH
- multipotentní kmenové buňky cytologie účinky léků MeSH
- pupečník cytologie MeSH
- transformované buněčné linie MeSH
- tvar buňky účinky léků MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Hypotonic solution alters ion channel activity, but little attention has been paid to voltage-dependent sodium channels. The aim of this study was to investigate the effects of hypotonic solution on transient sodium currents (INaT) and persistent sodium currents (INaP). We also explored whether the intracellular signal transduction systems participated in the hypotonic modifications of sodium currents. INaT and INaP were recorded by means of whole-cell patch-clamp technique in isolated rat ventricular myocytes. Our results revealed that hypotonic solution reduced INaT and simultaneously augmented INaP with the occurrence of interconversion between INaT and INaP. Hypotonic solution shifted steady-state inactivation to a more negative potential, prolonged the time of recovery from inactivation, and enhanced intermediate inactivation (IIM). Ruthenium red (RR, inhibitor of TRPV4), bisindolylmaleimide VI (BIM, inhibitor of PKC), Kn-93 (inhibitor of Ca/CaMKII) and BAPTA (Ca2+-chelator) inhibited the effects of hypotonic solution on INaT and INaP. Therefore we conclude that hypotonic solution inhibits INaT, enhances INaP and IIM with the effects being reversible. TRPV4 and intracellular Ca2+, PKC and Ca/CaMKII participate in the hypotonic modifications of sodium currents.
- MeSH
- benzylaminy farmakologie MeSH
- chelátory farmakologie MeSH
- EGTA analogy a deriváty farmakologie MeSH
- financování organizované MeSH
- hypotonické roztoky MeSH
- indoly farmakologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- kardiomyocyty metabolismus účinky léků MeSH
- kationtové kanály TRPV antagonisté a inhibitory metabolismus MeSH
- kinetika MeSH
- krysa rodu rattus MeSH
- maleimidy farmakologie MeSH
- membránové potenciály MeSH
- metoda terčíkového zámku MeSH
- proteinkinasa C antagonisté a inhibitory metabolismus MeSH
- proteinkinasa závislá na vápníku a kalmodulinu typ 2 antagonisté a inhibitory metabolismus MeSH
- rutheniová červeň farmakologie MeSH
- signální transdukce účinky léků MeSH
- sodík metabolismus MeSH
- srdeční komory cytologie metabolismus účinky léků MeSH
- sulfonamidy farmakologie MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
The vitamin E analogue alpha-tocopheryl succinate (alpha-TOS) is an efficient anti-cancer drug. Improved efficacy was achieved through the synthesis of alpha-tocopheryl maleamide (alpha-TAM), an esterase-resistant analogue of alpha-tocopheryl maleate. In vitro tests demonstrated significantly higher cytotoxicity of alpha-TAM towards cancer cells (MCF-7, B16F10) compared to alpha-TOS and other analogues prone to esterase-catalyzed hydrolysis. However, in vitro models demonstrated that alpha-TAM was cytotoxic to non-malignant cells (e.g. lymphocytes and bone marrow progenitors). Thus we developed lyophilized liposomal formulations of both alpha-TOS and alpha-TAM to solve the problem with cytotoxicity of free alpha-TAM (neurotoxicity and anaphylaxis), as well as the low solubility of both drugs. Remarkably, neither acute toxicity nor immunotoxicity implicated by in vitro tests was detected in vivo after application of liposomal alpha-TAM, which significantly reduced the growth of cancer cells in hollow fiber implants. Moreover, liposomal formulation of alpha-TAM and alpha-TOS each prevented the growth of tumours in transgenic FVB/N c-neu mice bearing spontaneous breast carcinomas. Liposomal formulation of alpha-TAM demonstrated anti-cancer activity at levels 10-fold lower than those of alpha-TOS. Thus, the liposomal formulation of alpha-TAM preserved its strong anti-cancer efficacy while eliminating the in vivo toxicity found of the free drug applied in DMSO. Liposome-based targeted delivery systems for analogues of vitamin E are of interest for further development of efficient and safe drug formulations for clinical trials.
- MeSH
- alfa-tokoferol analogy a deriváty aplikace a dávkování farmakologie MeSH
- antitumorózní látky aplikace a dávkování farmakologie MeSH
- chemie farmaceutická MeSH
- lidé MeSH
- liposomy MeSH
- maleimidy aplikace a dávkování farmakologie MeSH
- melanom experimentální farmakoterapie patologie MeSH
- myši inbrední BALB C MeSH
- myši inbrední C57BL MeSH
- myši transgenní MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory prsu farmakoterapie patologie MeSH
- polyethylenglykoly aplikace a dávkování farmakologie MeSH
- vitamin E analogy a deriváty aplikace a dávkování farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
- srovnávací studie MeSH
The effect of protein kinase C (PKC) inhibitors on porcine oocyte activation by calcium ionophore A23187 was studied. Calcium ionophore applied in a 50 microM concentration for 10 min induced activation in 74% of oocytes matured in vitro. When the ionophore-treated oocytes were exposed to the effect of bisindolylmaleimide I, which inhibits calcium-dependent PKC isotypes (PKC-alpha, -beta(I), -beta(II), -gamma,) and calcium-independent PKC isotypes (PKC-delta, -epsilon), the portion of activated oocytes decreased (at a concentration of 100 nM, 2% of the oocytes were activated). Go6976, the inhibitor of calcium-dependent PKC isotypes PKC-alpha, -beta(I) did not prevent the action of the oocytes treated with calcium ionophore in concentrations from 1 to 100 microM. The inhibitor of PKC-beta(I) and beta(II) isotypes, hispidin, in a concentration of 2 microM-2 mM, was not effective either. The inhibitor of PKC-delta isotype, rottlerin, suppressed activation of the oocytes by calcium ionophore (no oocyte was activated at 10 microM concentration). The PKC-delta isotype in matured porcine oocytes, studied by Western blot analysis, appeared as non-truncated PKC-delta of 77.5 kDa molecular weight, on the one hand, and as truncated PKC-delta, which was present in the form of a doublet of approximately 62.5 and 68 kDa molecular weight, on the other hand. On the basis of these results, it can be supposed that PKC participates in the regulation of processes associated with oocyte activation. Calcium-dependent PKC-alpha, -beta isotypes do not seem to play any significant role in calcium activation. The activation seems to depend on the activity of the calcium-independent PKC-delta isoform.
- MeSH
- acetofenony farmakologie MeSH
- benzopyrany farmakologie MeSH
- calcimycin farmakologie MeSH
- indoly farmakologie MeSH
- inhibitory proteinkinas farmakologie MeSH
- ionofory farmakologie MeSH
- izoenzymy antagonisté a inhibitory fyziologie klasifikace MeSH
- karbazoly farmakologie MeSH
- maleimidy farmakologie MeSH
- oocyty fyziologie účinky léků MeSH
- prasata fyziologie MeSH
- proteinkinasa C antagonisté a inhibitory fyziologie klasifikace MeSH
- pyrony farmakologie MeSH
- vápník fyziologie MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- kazuistiky MeSH
