Metallothioneins (MTs), low molecular mass cysteine-rich proteins, which are able to bind up to 20 monovalent and up to 7 divalent heavy metal ions are widely studied due to their functions in detoxification of metals, scavenging free radicals and cells protection against the oxidative stress. It was found that the loss of the protective effects of MT leads to an escalation of pathogenic processes and carcinogenesis. The most extensive area is MTs expression for oncological applications, where the information about gene patterns is helpful for the identification biological function, resistance to drugs and creating the correct chemotherapy. In other medical applications the effect of oxidative stress to cell lines exposed to heavy metals and hydrogen peroxide is studied as well as influence of drugs and cytokines on MTs expression and MTs expression in the adipose tissue. The precise detection of low metallothionein concentrations and its isoforms is necessary to understand the connection between quantity and isoforms of MTs to size, localization and type of cancer. This information is necessary for well-timed therapy and increase the chance to survival. Microarray chips appear as good possibility for finding all information about expression of MTs genes and isoforms not only in cancer, but also in other diseases, especially diabetes, obesity, cardiovascular diseases, ageing, osteoporosis, psychiatric disorders and as the effects of toxic drugs and pollutants, which is discussed in this review.
- Klíčová slova
- Drug, Gene expression, Metallothionein, Microarray, Resistance, Tumour diseases,
- MeSH
- lidé MeSH
- metalothionein analýza genetika metabolismus MeSH
- mikročipová analýza metody MeSH
- nádory diagnóza genetika metabolismus MeSH
- oxidační stres fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- metalothionein MeSH
Interaction between semiconductor nanocrystals, cadmium telluride quantum dots (CdTe QDs) capped with mercaptosuccinic acid (MSA) and metallothionein (MT) was investigated. MSA-capped CdTe QDs were synthesized in aqueous solution. Mixture of MT and MSA-capped CdTe QDs has been investigated by various analytical methods as follows: tris-tricine gel electrophoresis, fluorescence evaluation and electrochemical detection of catalysed hydrogen evolution. The obtained results demonstrate that MSA-capped CdTe QDs and MT do not create firmly bound stabile complex. However, weak electrostatic interactions contribute to the interaction of MT with MSA-capped CdTeQDs. It can be concluded that QDs size influences the QDs and MT interaction. The smallest QDs had the highest affinity to MT and vice versa.
- Klíčová slova
- Complex, Interaction, Metallothionein, Quantum dot,
- MeSH
- elektrochemické techniky MeSH
- fluorescenční spektrometrie MeSH
- králíci MeSH
- kvantové tečky * MeSH
- metalothionein chemie MeSH
- molekulární struktura MeSH
- nanočástice * MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- metalothionein MeSH
The present study suggests and describes the application of a delivery system for antisense oligonucleotides against mRNA encoding estrogen receptor proteins α and β. The delivery system is composed of a cationic liposome envelope containing 17β-estradiol (E2) in its structure. Cationic liposomes protect cargo against the extracellular matrix, and E2 can increase its shuttling efficiency into cells. Using MCF-7 cells derived from estrogen receptor-positive ductal carcinoma, treatment with liposomes against ERα was found to decrease MCF-7 proliferation, and importantly the application of both the antisense against ERα and β exhibited an antiproliferative effect expressed as cell viability. Using qRT-PCR, it was shown that MT1A, NF-κB1 and K-ras genes, but not TFF1, were downregulated using E2-based liposomes (evaluated at P=0.05). Further indicators of oxidative stress were employed to assess the effect on treatment efficiency. Glutathione (GSH/GSSG redox ratio), metallothionein (MT) and malondialdehyde (MDA) confirmed a positive effect of antisense therapy resulting in their decreased levels in the MCF-7 cells. Based on these data, we suggest that E2-based liposomes offer sufficient transfer efficiency and moreover, due to the effect on NF-κB1, MT and GSH, tumor cells can be chemosensitized to increase treatment effectiveness.
- MeSH
- estradiol metabolismus MeSH
- genetická terapie metody MeSH
- glutathion chemie MeSH
- koncentrace vodíkových iontů MeSH
- kyslík chemie MeSH
- lékové transportní systémy * MeSH
- lidé MeSH
- liposomy chemie MeSH
- malondialdehyd chemie MeSH
- metalothionein chemie MeSH
- MFC-7 buňky MeSH
- mikroskopie fázově kontrastní MeSH
- nádorové buněčné linie MeSH
- nádory prsu metabolismus MeSH
- oxidace-redukce MeSH
- oxidační stres MeSH
- proliferace buněk MeSH
- protinádorové látky chemie MeSH
- receptory pro estrogeny metabolismus MeSH
- regulace genové exprese MeSH
- viabilita buněk MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- estradiol MeSH
- glutathion MeSH
- kyslík MeSH
- liposomy MeSH
- malondialdehyd MeSH
- metalothionein MeSH
- protinádorové látky MeSH
- receptory pro estrogeny MeSH
Modification of carbon materials, especially graphene-based materials, has wide applications in electrochemical detection such as electrochemical lab-on-chip devices. A glassy carbon electrode (GCE) modified with chemically alternated graphene oxide was used as a working electrode (glassy carbon modified by graphene oxide with sulphur containing compounds and Nafion) for detection of nucleobases in hydrolysed samples (HCl pH = 2.9, 100 °C, 1 h, neutralization by NaOH). It was found out that modification, especially with trithiocyanuric acid, increased the sensitivity of detection in comparison with pure GCE. All processes were finally implemented in a microfluidic chip formed with a 3D printer by fused deposition modelling technology. As a material for chip fabrication, acrylonitrile butadiene styrene was chosen because of its mechanical and chemical stability. The chip contained the one chamber for the hydrolysis of the nucleic acid and another for the electrochemical detection by the modified GCE. This chamber was fabricated to allow for replacement of the GCE.
- Publikační typ
- časopisecké články MeSH
In this study, we focused on the effect of heavy metal ions in resistant strains of gram-positive bacteria Staphylococcus aureus using biochemical methods and mass spectrometry. Five nitrate solutions of heavy metals (Ag(+), Cu(2+), Cd(2+), Zn(2+) and Pb(2+)) were used to create S. aureus resistant strains. Biochemical changes of resistant strains in comparison with the non-resistant control strain of S. aureus were observed by microbiological (measuring - growth curves and inhibition zones) and spectrophotometric methods (antioxidant activity and alaninaminotransferase, aspartateaminotransferase, alkaline phosphatase, γ-glutamyltransferase activities). Mass spectrometry was employed for the qualitative analysis of the samples (changes in S. aureus protein composition) and for the identification of the strains database MALDI Biotyper was employed. Alterations, in terms of biochemical properties and protein composition, were observed in resistant strains compared to non-resistant control strain. Our results describe the possible option for the analysis of S. aureus resistant strains and may thus serve as a support for monitoring of changes in genetic information caused by the forming of resistance to heavy metals.
- Klíčová slova
- Heavy metals, Mass spectrometry, Resistance, S. aureus,
- MeSH
- hmotnostní spektrometrie MeSH
- ionty metabolismus farmakologie MeSH
- kovy metabolismus farmakologie MeSH
- metabolom MeSH
- metabolomika metody MeSH
- metalothionein chemie metabolismus MeSH
- mikrobiální viabilita účinky léků MeSH
- RNA ribozomální genetika MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Staphylococcus aureus účinky léků genetika růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ionty MeSH
- kovy MeSH
- metalothionein MeSH
- RNA ribozomální MeSH
Nanoparticles have gained increasing interest in medical and in vivo applications. Metallothionein (MT) is well known as a maintainer of metal ions balance in intracellular space. This is due to high affinity of this protein to any reactive species including metals and reactive oxygen species. The purpose of this study was to determine the metallothionein-quantum dots interactions that were investigated by spectral and electrochemical techniques. CuS, CdS, PbS, and CdTe quantum dots (QDs) were analysed. The highest intensity was shown for CdTe, than for CdS measured by fluorescence. These results were supported by statistical analysis and considered as significant. Further, these interactions were analysed using gel electrophoresis, where MT aggregates forming after interactions with QDs were detected. Using differential pulse voltammetry Brdicka reaction, QDs and MT were studied. This method allowed us to confirm spectral results and, moreover, to observe the changes in MT structure causing new voltammetric peaks called X and Y, which enhanced with the prolonged time of interaction up to 6 h.
- Klíčová slova
- Electrochemical detection, Interaction, Metallothionein, Quantum dots,
- MeSH
- elektrochemické techniky MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- fluorescence MeSH
- králíci MeSH
- kvantové tečky metabolismus MeSH
- metalothionein metabolismus MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- metalothionein MeSH
There is an arising and concerning issue in the field of bacterial resistance, which is confirmed by the number of deaths associated with drug-resistant bacterial infections. The aim of this study was to compare the effects of antibiotics on Staphylococcus aureus non-resistant strain and strains resistant to cadmium or lead ions. Metal resistant strains were created by the gradual addition of 2 mM solution of metal ions (cadmium or lead) to the S. aureus culture. An increasing antimicrobial effect of ampicillin, streptomycin, penicillin and tetracycline (0, 10, 25, 50, 75, 150, 225 and 300 µM) on the resistant strains was observed using a method of growth curves. A significant growth inhibition (compared to control) of cadmium resistant cells was observed in the presence of all the four different antibiotics. On the other hand, the addition of streptomycin and ampicillin did not inhibit the growth of lead resistant strain. Other antibiotics were still toxic to the bacterial cells. Significant differences in the morphology of cell walls were indicated by changes in the cell shape. Our data show that the presence of metal ions in the urban environment may contribute to the development of bacterial strain resistance to other substances including antibiotics, which would have an impact on public health.
- MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální léková rezistence * MeSH
- kadmium farmakologie MeSH
- mikrobiální testy citlivosti MeSH
- olovo farmakologie MeSH
- Staphylococcus aureus účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- kadmium MeSH
- olovo MeSH
Interactions of silver phosphate nanoparticles (SPNPs) and selenium nanoparticles (SeNPs) with Staphylococcus aureus cultures have been studied at the cellular, molecular and protein level. Significant antibacterial effects of both SPNPs and SeNPs on S. aureus were observed. At a concentration of 300 μM, SPNPs caused 37.5% inhibition of bacterial growth and SeNPs totally inhibited bacterial growth. As these effects might have been performed due to the interactions of nanoparticles with DNA and proteins, the interaction of SPNPs or SeNPs with the amplified zntR gene was studied. The presence of nanoparticles decreased the melting temperatures of the nanoparticle complexes with the zntR gene by 23% for SeNPs and by 12% for SPNPs in comparison with the control value. The concentration of bacterial metallothionein was 87% lower in bacteria after application of SPNPs (6.3 μg mg(-1) protein) but was increased by 29% after addition of SeNPs (63 μg mg(-1) protein) compared with the S. aureus control (49 μg mg(-1) protein). Significant antimicrobial effects of the nanoparticles on bacterial growth and DNA integrity provide a promising approach to reducing the risk of bacterial infections that cannot be controlled by the usual antibiotic treatments.
- Klíčová slova
- antimicrobial effect, growth, inhibition, nanotechnology,
- MeSH
- antibakteriální látky farmakologie MeSH
- fosfáty farmakologie MeSH
- mikrobiální testy citlivosti MeSH
- nanočástice * MeSH
- selen farmakologie MeSH
- sloučeniny stříbra farmakologie MeSH
- stafylokokové infekce prevence a kontrola MeSH
- Staphylococcus aureus účinky léků růst a vývoj MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- fosfáty MeSH
- selen MeSH
- silver phosphate MeSH Prohlížeč
- sloučeniny stříbra MeSH
Ellipticine (5,11-dimethyl-6H-pyrido[4,3-b]carbazole) is an alkaloid that has been isolated from plants of an Apocynaceae family. It is one of the simplest naturally occurring alkaloids with a planar structure. Over the past decades, ellipticine became a very promising antitumor agent. Interaction with DNA is one of the most studied ellipticine effects on cell division. This phenomenon is not clearly explained so far. In our experiments we studied interaction of ellipticine with single-stranded and double-stranded oligonucleotides by electrochemical methods on mercury electrode. Differential pulse voltammetry was applied for ellipticine (Elli) and CA peak detection. Square wave voltammetry was applied for G peak detection. The effect of the interaction time and ellipticine concentrations on interactions of ellipticine with single- and double-stranded oligonucleotides was tested too.
- MeSH
- Apocynaceae chemie MeSH
- elektrochemické techniky MeSH
- elipticiny chemie MeSH
- oligonukleotidy chemie MeSH
- protinádorové látky chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- elipticiny MeSH
- ellipticine MeSH Prohlížeč
- oligonukleotidy MeSH
- protinádorové látky MeSH
INTRODUCTION: Breast cancer is one of the most common types of cancer in women. One of the genes that were found mutated in breast cancer is casein kinase 1 epsilon (CK1epsilon). Because CK1epsilon is a crucial regulator of the Wnt signaling cascades, we determined how these CK1epsilon mutations interfere with the Wnt pathway and affect the behavior of epithelial breast cancer cell lines. METHODS: We performed in silico modeling of various mutations and analyzed the kinase activity of the CK1epsilon mutants both in vitro and in vivo. Furthermore, we used reporter and small GTPase assays to identify how mutation of CK1epsilon affects different branches of the Wnt signaling pathway. Based on these results, we employed cell adhesion and cell migration assays in MCF7 cells to demonstrate a crucial role for CK1epsilon in these processes. RESULTS: In silico modeling and in vivo data showed that autophosphorylation at Thr 44, a site adjacent to the breast cancer point mutations in the N-terminal lobe of human CK1epsilon, is involved in positive regulation of the CK1epsilon activity. Our data further demonstrate that, in mammalian cells, mutated forms of CK1epsilon failed to affect the intracellular localization and phosphorylation of Dvl2; we were able to demonstrate that CK1epsilon mutants were unable to enhance Dvl-induced TCF/LEF-mediated transcription, that CK1epsilon mutants acted as loss-of-function in the Wnt/beta-catenin pathway, and that CK1epsilon mutants activated the noncanonical Wnt/Rac-1 and NFAT pathways, similar to pharmacological inhibitors of CK1. In line with these findings, inhibition of CK1 promoted cell migration as well as decreased cell adhesion and E-cadherin expression in the breast cancer-derived cell line MCF7. CONCLUSIONS: In summary, these data suggest that the mutations of CK1epsilon found in breast cancer can suppress Wnt/beta-catenin as well as promote the Wnt/Rac-1/JNK and Wnt/NFAT pathways, thus contributing to breast cancer development via effects on cell adhesion and migration. In terms of molecular mechanism, our data indicate that the breast cancer point mutations in the N-terminal lobe of CK1epsilon, which are correlated with decreased phosphorylation activities of mutated forms of CK1epsilon both in vitro and in vivo, interfere with positive autophosphorylation at Thr 44.
- MeSH
- beta-katenin metabolismus MeSH
- buněčná adheze MeSH
- duktální karcinom prsu genetika metabolismus patologie MeSH
- fosforylace MeSH
- imunoprecipitace MeSH
- kaseinkinasa Iepsilon chemie genetika metabolismus MeSH
- konformace proteinů MeSH
- lidé MeSH
- MAP kinasa-kinasa 4 metabolismus MeSH
- messenger RNA genetika MeSH
- mutace genetika MeSH
- nádorové buněčné linie MeSH
- nádory prsu genetika metabolismus patologie MeSH
- pohyb buněk * MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proliferace buněk MeSH
- proteiny Wnt metabolismus MeSH
- rac1 protein vázající GTP metabolismus MeSH
- transkripční faktory NFATC metabolismus MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- beta-katenin MeSH
- kaseinkinasa Iepsilon MeSH
- MAP kinasa-kinasa 4 MeSH
- messenger RNA MeSH
- proteiny Wnt MeSH
- rac1 protein vázající GTP MeSH
- RAC1 protein, human MeSH Prohlížeč
- transkripční faktory NFATC MeSH
