- MeSH
- akutní promyelocytární leukemie * diagnóza farmakoterapie krev patologie MeSH
- buňky kostní dřeně patologie MeSH
- časná diagnóza MeSH
- indukční chemoterapie MeSH
- kombinovaná farmakoterapie MeSH
- konsolidační chemoterapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- oxid arsenitý ekonomika terapeutické užití MeSH
- pancytopenie etiologie MeSH
- protinádorové látky aplikace a dávkování MeSH
- tretinoin aplikace a dávkování MeSH
- vyšetřování kostní dřeně MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- akutní promyelocytární leukemie * farmakoterapie krev patologie MeSH
- dospělí MeSH
- indukovaný potrat MeSH
- klinické rozhodování MeSH
- kombinovaná farmakoterapie MeSH
- konsolidační chemoterapie MeSH
- lidé MeSH
- oxid arsenitý ekonomika terapeutické užití MeSH
- protinádorové látky aplikace a dávkování MeSH
- těhotenství * MeSH
- tretinoin aplikace a dávkování MeSH
- zapojení pacienta MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- těhotenství * MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Dendrimers, which are considered as one of the most promising tools in the field of nanobiotechnology due to their structural organization, showed a great potential in gene therapy, drug delivery, medical imaging and as antimicrobial and antiviral agents. This article is devoted to study interactions between new carbosilane-based metallodendrimers containing ruthenium and anti-cancer small interfering RNA (siRNA). Formation of complexes between anti-cancer siRNAs and Ru-based carbosilane dendrimers was evaluated by transmission electron microscopy, circular dichroism and fluorescence. The zeta-potential and the size of dendriplexes were determined by dynamic light scattering. The internalization of dendriplexes were estimated using HL-60 cells. Results show that ruthenium dendrimers associated with anticancer siRNA have the ability to deliver siRNA as non-viral vectors into the cancer cells. Moreover, dendrimers can protect siRNA against nuclease degradation. Nevertheless, further research need to be performed to examine the therapeutic potential of ruthenium dendrimers as well as dendrimers complexed with siRNA and anticancer drugs towards cancer cells.
- MeSH
- akutní promyelocytární leukemie farmakoterapie metabolismus patologie MeSH
- cirkulární dichroismus MeSH
- dendrimery aplikace a dávkování chemie metabolismus MeSH
- dynamický rozptyl světla MeSH
- fyziologická absorpce MeSH
- HL-60 buňky MeSH
- interkalátory aplikace a dávkování chemie metabolismus MeSH
- lidé MeSH
- malá interferující RNA aplikace a dávkování chemie metabolismus ultrastruktura MeSH
- molekulární konformace MeSH
- molekulární modely * MeSH
- molekulární struktura MeSH
- povrchové vlastnosti MeSH
- protinádorové látky aplikace a dávkování chemie metabolismus MeSH
- RNA interference MeSH
- ruthenium aplikace a dávkování chemie metabolismus MeSH
- silany chemie metabolismus MeSH
- simulace molekulární dynamiky MeSH
- stabilita léku MeSH
- stabilita RNA MeSH
- transmisní elektronová mikroskopie MeSH
- velikost částic MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
We studied epigenetics, distribution pattern, kinetics, and diffusion of proteins recruited to spontaneous and γ-radiation-induced DNA lesions. We showed that PML deficiency leads to an increased number of DNA lesions, which was accompanied by changes in histone signature. In PML wt cells, we observed two mobile fractions of 53BP1 protein with distinct diffusion in spontaneous lesions. These protein fractions were not detected in PML-deficient cells, characterized by slow-diffusion of 53BP1. Single particle tracking analysis revealed limited local motion of 53BP1 foci in PML double null cells and local motion 53BP1 foci was even more reduced after γ-irradiation. However, radiation did not change co-localization between 53BP1 nuclear bodies and interchromatin granule-associated zones (IGAZs), nuclear speckles, or chromocenters. This newly observed interaction pattern imply that 53BP1 protein could be a part of not only DNA repair, but also process mediated via components accumulated in IGAZs, nuclear speckles, or paraspeckles. Together, PML deficiency affected local motion of 53BP1 nuclear bodies and changed composition and a number of irradiation-induced foci. J. Cell. Biochem. 117: 2583-2596, 2016. © 2016 Wiley Periodicals, Inc.
- MeSH
- 53BP1 metabolismus MeSH
- akutní promyelocytární leukemie metabolismus patologie radioterapie MeSH
- fluorescenční protilátková technika MeSH
- intranukleární inkluzní tělíska metabolismus patologie účinky záření MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- nádorové buňky kultivované MeSH
- oprava DNA fyziologie účinky záření MeSH
- poškození DNA fyziologie účinky záření MeSH
- vztah dávky záření a odpovědi MeSH
- western blotting MeSH
- záření gama škodlivé účinky MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
HL-60 cancer cells were treated with a series of novel acridine derivatives (derivatives 1-4) in order to test the compounds' ability to inhibit both cancer cell growth and topoisomerase I and II activity. Binding studies of derivatives 1-4 with calf thymus DNA were also performed using a number of techniques (UV-Vis and fluorescence spectroscopy, thermal denaturation, linear dichroism and viscometry) to determine the nature of the interaction between the compounds and ctDNA. The binding constants for the complexes of the studied acridine derivatives with DNA were calculated from UV-Vis spectroscopic titrations (K=3.1×10(4)-2.0×10(3)M(-1)). Some of the compounds showed a strong inhibitory effect against Topo II at the relatively low concentration of 5μM. Topo I/II inhibition mode assays were also performed and verified that the novel compounds are topoisomerase suppressors rather than poisons. The biological activities of derivatives were studied using MTT assay and flow cytometric methods (detection of mitochondrial membrane potential, measurement of cell viability) after 24 and 48h incubation. The ability of derivatives to impair cell proliferation was tested by an analysis of cell cycle distribution.
- MeSH
- akridiny chemická syntéza metabolismus farmakologie MeSH
- akutní promyelocytární leukemie farmakoterapie enzymologie patologie MeSH
- časové faktory MeSH
- denaturace nukleových kyselin MeSH
- DNA-topoisomerasy I metabolismus MeSH
- DNA-topoisomerasy typu II metabolismus MeSH
- DNA chemie metabolismus MeSH
- farmaceutická chemie MeSH
- farmaceutická technologie metody MeSH
- fluorescenční spektrometrie MeSH
- HL-60 buňky MeSH
- inhibitory topoisomerasy I chemická syntéza metabolismus farmakologie MeSH
- inhibitory topoisomerasy II chemická syntéza metabolismus farmakologie MeSH
- konformace nukleové kyseliny MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- lidé MeSH
- membránový potenciál mitochondrií účinky léků MeSH
- proliferace buněk účinky léků MeSH
- spektrofotometrie ultrafialová MeSH
- viabilita buněk účinky léků MeSH
- viskozita MeSH
- vysoká teplota MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- akutní promyelocytární leukemie * diagnóza farmakoterapie genetika patologie MeSH
- antracykliny aplikace a dávkování MeSH
- arsenitany aplikace a dávkování MeSH
- fúzní onkogenní proteiny genetika metabolismus MeSH
- kombinovaná farmakoterapie metody MeSH
- lidé MeSH
- nádorová transformace buněk genetika MeSH
- polymerázová řetězová reakce s reverzní transkripcí metody MeSH
- prekurzorové buňky granulocytů patologie MeSH
- regulace genové exprese u nádorů MeSH
- tretinoin aplikace a dávkování MeSH
- udržovací chemoterapie metody MeSH
- Check Tag
- lidé MeSH
The histamine H4 receptor regulates the inflammatory response. However, it is not known whether this receptor has a functional role in human neutrophils. We found that fMLP (1 μM), but not histamine (0.1-1 μM), induced Mac-1-dependent adhesion, polarization, and degranulation (release of lactoferrin). A pretreatment of neutrophils with histamine (0.001-1 μM) or JNJ 28610244 (0.1-10 μM), a specific H4 receptor agonist, led to inhibition of degranulation. Total inhibition of degranulation was obtained with 0.1 μM histamine and 10 μM JNJ 28610244. Furthermore, such inhibition by histamine of degranulation was reversed by JNJ 7777120 and JNJ 28307474, two selective H4 receptor antagonists. However, neither histamine nor the H4 receptor agonist JNJ 28610244 prevented fMLP-induced, Mac-1-dependent adhesion, indicating that the H4 receptor may block signals emanating from Mac-1-controlling degranulation. Likewise, engagement of the H4 receptor by the selective agonist JNJ 28610244 blocked Mac-1-dependent activation of p38 MAPK, the kinase that controls neutrophil degranulation. We also show expression of the H4 receptor at the mRNA level in ultrapure human neutrophils and myeloid leukemia PLB-985 cells. We concluded that engagement of this receptor by selective H4 receptor agonists may represent a good, therapeutic approach to accelerate resolution of inflammation.
- MeSH
- akutní promyelocytární leukemie patologie MeSH
- antigen-1 spojený s lymfocytární funkcí chemie MeSH
- buněčná adheze účinky léků fyziologie MeSH
- cytochalasin B farmakologie MeSH
- degranulace buněk * účinky léků MeSH
- fibrinogen MeSH
- histamin farmakologie MeSH
- indoly farmakologie MeSH
- konformace proteinů účinky léků MeSH
- kultivované buňky MeSH
- lidé MeSH
- makrofágový antigen 1 fyziologie MeSH
- MAP kinasový signální systém účinky léků MeSH
- messenger RNA biosyntéza genetika MeSH
- mitogenem aktivované proteinkinasy p38 fyziologie MeSH
- N-formylmethionin-leucyl-fenylalanin farmakologie MeSH
- nádorové buněčné linie MeSH
- neutrofily účinky léků fyziologie MeSH
- oximy farmakologie MeSH
- piperaziny farmakologie MeSH
- piperidiny farmakologie MeSH
- pyridiny farmakologie MeSH
- receptory histaminu fyziologie MeSH
- receptory spřažené s G-proteiny agonisté antagonisté a inhibitory fyziologie MeSH
- tvar buňky účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We compared the effects of inhibitors of kinases ATM (KU55933) and ATR (VE-821) (incubated for 30 min before irradiation) on the radiosensitization of human promyelocyte leukaemia cells (HL-60), lacking functional protein p53. VE-821 reduces phosphorylation of check-point kinase 1 at serine 345, and KU55933 reduces phosphorylation of check-point kinase 2 on threonine 68 as assayed 4 h after irradiation by the dose of 6 Gy. Within 24 h after gamma-irradiation with a dose of 3 Gy, the cells accumulated in the G2 phase (67 %) and the number of cells in S phase decreased. KU55933 (10 μM) did not affect the accumulation of cells in G2 phase and did not affect the decrease in the number of cells in S phase after irradiation. VE-821 (2 and 10 μM) reduced the number of irradiated cells in the G2 phase to the level of non-irradiated cells and increased the number of irradiated cells in S phase, compared to irradiated cells not treated with inhibitors. In the 144 h interval after irradiation with 3 Gy, there was a considerable induction of apoptosis in the VE-821 group (10 μM). The repair of the radiation damage, as observed 72 h after irradiation, was more rapid in the group exposed solely to irradiation and in the group treated with KU55933 (80 and 77 % of cells, respectively, were free of DSBs), whereas in the group incubated with 10 μM VE-821, there were only 61 % of cells free of DSBs. The inhibition of kinase ATR with its specific inhibitor VE-821 resulted in a more pronounced radiosensitizing effect in HL-60 cells as compared to the inhibition of kinase ATM with the inhibitor KU55933. In contrast to KU55933, the VE-821 treatment prevented HL-60 cells from undergoing G2 cell cycle arrest. Taken together, we conclude that the ATR kinase inhibition offers a new possibility of radiosensitization of tumour cells lacking functional protein p53.
- MeSH
- akutní promyelocytární leukemie patologie MeSH
- apoptóza účinky léků MeSH
- ATM protein antagonisté a inhibitory MeSH
- HL-60 buňky MeSH
- inhibitory proteinkinas farmakologie MeSH
- kontrolní body fáze G2 buněčného cyklu účinky léků MeSH
- lidé MeSH
- morfoliny farmakologie MeSH
- oprava DNA účinky léků MeSH
- pyraziny farmakologie MeSH
- pyrony farmakologie MeSH
- sulfony farmakologie MeSH
- tolerance záření účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We used hybrid detectors (HyDs) to monitor the trajectories and interactions of promyelocytic leukemia (GFP-PML) nuclear bodies (NBs) and mCherry-53BP1-positive DNA lesions. 53BP1 protein accumulates in NBs that occur spontaneously in the genome or in γ-irradiation-induced foci. When we induced local DNA damage by ultraviolet irradiation, we also observed accumulation of 53BP1 proteins into discrete bodies, instead of the expected dispersed pattern. In comparison with photomultiplier tubes, which are used for standard analysis by confocal laser scanning microscopy, HyDs significantly eliminated photobleaching of GFP and mCherry fluorochromes during image acquisition. The low laser intensities used for HyD-based confocal analysis enabled us to observe NBs for the longer time periods, necessary for studies of the trajectories and interactions of PML and 53BP1 NBs. To further characterize protein interactions, we used resonance scanning and a novel bioinformatics approach to register and analyze the movements of individual PML and 53BP1 NBs. The combination of improved HyD-based confocal microscopy with a tailored bioinformatics approach enabled us to reveal damage-specific properties of PML and 53BP1 NBs.
- MeSH
- akutní promyelocytární leukemie metabolismus patologie MeSH
- časosběrné zobrazování MeSH
- DNA chemie metabolismus MeSH
- intracelulární signální peptidy a proteiny metabolismus MeSH
- intranukleární inkluzní tělíska metabolismus ultrastruktura MeSH
- konfokální mikroskopie přístrojové vybavení metody MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- poškození DNA MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Tumor suppressor PML is induced under viral and genotoxic stresses by interferons and JAK-STAT signaling. However, the mechanism responsible for its cell type-specific regulation under non-stimulated conditions is poorly understood. To analyze the variation of PML expression, we utilized three human cell types, BJ fibroblasts and HeLa and U2OS cell lines, each with a distinct PML expression pattern. Analysis of JAK-STAT signaling in the three cell lines revealed differences in levels of activated STAT3 but not STAT1 correlating with PML mRNA and protein levels. RNAi-mediated knockdown of STAT3 decreased PML expression; both STAT3 level/activity and PML expression relied on IL6 secreted into culture media. We mapped the IL6-responsive sequence to an ISRE(-595/-628) element of the PML promoter. The PI3K/Akt/NFκB branch of IL6 signaling showed also cell-type dependence, being highest in BJ, intermediate in HeLa, and lowest in U2OS cells and correlated with IL6 secretion. RNAi-mediated knockdown of NEMO (NF-κ-B essential modulator), a key component of NFκB activation, suppressed NFκB targets LMP2 and IRF1 together with STAT3 and PML. Combined knockdown of STAT3 and NEMO did not further promote PML suppression, and it can be bypassed by exogenous IL6, indicating the NF-κB pathway acts upstream of JAK-STAT3 through induction of IL6. Our results indicate that the cell type-specific activity of IL6 signaling pathways governs PML expression under unperturbed growth conditions. As IL6 is induced in response to various viral and genotoxic stresses, this cytokine may regulate autocrine/paracrine induction of PML under these pathophysiological states as part of tissue adaptation to local stress.
- MeSH
- akutní promyelocytární leukemie metabolismus patologie MeSH
- chromatinová imunoprecipitace MeSH
- DNA primery MeSH
- fluorescenční protilátková technika nepřímá MeSH
- HeLa buňky MeSH
- interleukin-6 metabolismus MeSH
- jaderné proteiny metabolismus MeSH
- Janus kinasy metabolismus MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- nádorové supresorové proteiny metabolismus MeSH
- sekvence nukleotidů MeSH
- signální transdukce MeSH
- transkripční faktory STAT metabolismus MeSH
- transkripční faktory metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
