Schiff base copper (II) complexes are known for their anticancer, antifungal, antiviral and anti‑inflammatory activities. The aim of the current study was to investigate biological effects of Schiff base Cu (II) complexes (0.001‑100 µmol/l)‑[Cu2(sal‑D, L‑glu)2(isoquinoline)2]·2C2H5OH (1), [Cu(sal‑5‑met‑L‑glu)(H2O)].H2O (2), [Cu(ethanol)2(imidazole)4][Cu2(sal‑D, L-glu)2(imidazole)2] (3), [Cu(sal‑D,L‑glu)(2‑methylimidazole)] (4) on the human colon carcinoma cells HT‑29, the mouse noncancerous cell line NIH‑3T3 and the human noncancerous fibroblast cell line VH10. The results suggested that Cu (II) complexes exhibit cytotoxic effects against the HT‑29 cell line, while complexes 3 and 4 were the most effective. Subsequent to 72 h of incubation, apoptosis was observed in the HT‑29 cells induced by Cu (II) complexes 1 (0.1, 1, 10 and 50 µmol/l), 2 (1, 10, 50 and 100 µmol/l), 3 (0.01, 1, 10 and 50 µmol/l) and 4 (0.01, 0.1, 1 and 10 µmol/l). The apoptotic pathways activated by the Cu (II) complexes were identified. The results indicated that complexes 2, 3 and 4 were able to induce the mitochondria‑dependent pathway of apoptosis in HT‑29 cells, while complex 1 was obsered to activate the extrinsic pathway of apoptosis. The levels of the anti‑apoptotic protein Bcl‑2 were reduced and those of the pro‑apoptotic protein Bax increased following treatment with complexes 2, 3 and 4. Complex 1 had no effect on Bax protein expression. Complexes 2 and 3 induced elevation of cytochrome c (cyt c), while complex 4 induced a time‑dependent elevation of cyt c levels. No cyt c was detected in HT‑29 cells exposed to complex 1, suggesting that Cu (II) complexes activated the extrinsic pathway of apoptosis. The results from the current study in addition to previous studies suggest that Schiff base Cu (II) complexes have potential as novel anticancer drugs.
- MeSH
- antitumorózní látky aplikace a dávkování MeSH
- apoptóza účinky léků MeSH
- buňky HT-29 MeSH
- lidé MeSH
- měď aplikace a dávkování MeSH
- mitochondrie účinky léků MeSH
- myši MeSH
- nádorové proteiny biosyntéza MeSH
- nádory tračníku farmakoterapie patologie MeSH
- proliferace buněk účinky léků MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- Schiffovy báze aplikace a dávkování MeSH
- signální transdukce účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Autophagy plays an important role in cancer cells. Targeting autophagy in cancer can provide new opportunities for drug development. METHODS: In this study we tested four Schiff base Cu(II) complexes against human breast cancer cells (MCF-7) and human non-cancerous cells (HEK-293T). We have tested their cytotoxic effect by evaluating IC50 using MTT test. To detect morphological changes of the actin fibers we have used fluorescent microscopy. To determine the type of cell death we used electrophoretic analysis and western blot analysis (protein LC3). RESULTS: IC50 values of the complexes increased with time of their influence, indicating acquired resistance of MCF-7 to the complexes. Healthy cells HEK-293T were not sensitive to the Cu(II) complexes. Compared with the control cells (cells without Cu(II) complexes) which were without morphological changes of actin fibers, Cu(II) complexes induced condensation and asymmetric conformational changes in actin filaments. To examine the type of cell death induced by the Cu(II) complexes we treated MCF-7 cells with Cu(II) complexes (1, 10, 50 and 100μmol/L) during a 72h incubation period. By electrophoresis we have not detected any DNA fragmentation. To determine whether Cu(II) complexes induced autophagy or necrotic cell death we used the western blot analysis. MCF-7 cells influenced with tested Cu(II) complexes produced LC3 protein after their 72h incubation indicating autophagy in MCF-7 cancer cells. CONCLUSIONS: Tested Schiff base copper (II) complexes have antiproliferative activity against cancer cells but not against healthy cells. They have induced autophagy in the cancer cell line MCF-7.
- MeSH
- autofagie účinky léků fyziologie MeSH
- HEK293 buňky MeSH
- lidé MeSH
- měď farmakologie toxicita MeSH
- MFC-7 buňky MeSH
- myši MeSH
- proliferace buněk účinky léků fyziologie MeSH
- Schiffovy báze farmakologie toxicita MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Statins (HMG-CoA reductase inhibitors) represent a major class of compounds for the treatment of hypercholesterolemia due to their ability to inhibit de novo cholesterol synthesis. In addition to their hypolipidemic effects, chemoprotective properties have been attributed to statins as well. These effects involve multiple mechanisms, which, however, are not known in detail. The aim of our study was to assess in non-malignant as well as cancer cells the impact of simvastatin on the amount of cytosolic lipid droplets (LDs) implicated in many biological processes including proliferation, inflammation, carcinogenesis, apoptosis, necrosis or growth arrest. METHODS: Human embryonic kidney cells HEK-293T and human pancreatic cancer cells MiaPaCa-2 were treated with simvastatin (6 and 12 muM) for 24 and 48 hours respectively. Neutral lipid probe Nile Red was used for detection of LDs by fluorescence microscopy. Cellular cholesterol content was determined by HPLC. Changes in expression of genes related to lipid metabolism in simvastatin-treated MiaPaCa-2 cells were examined by DNA microarray analysis. Validation of gene expression changes was performed using quantitative RT-PCR. RESULTS: The treatment of the cells with simvastatin increased their intracellular content of LDs in both non-malignant as well as cancer cells, partially due to the uptake of cholesterol and triacylglyceroles from medium; but in particular, due to enhanced synthesis of triacylglyceroles as proved by significant overexpression of genes related to de novo synthesis of triacylglyceroles and phospholipids. In addition, simvastatin also markedly influenced expression of genes directly affecting cell proliferation and signaling. CONCLUSIONS: Simvastatin treatment led to accumulation of cytosolic LDs within the examined cells, a phenomenon which might contribute to the antiproliferative effects of statins.
- MeSH
- cholesterol metabolismus MeSH
- HEK293 buňky účinky léků MeSH
- lidé MeSH
- lipidová tělíska * metabolismus účinky léků MeSH
- metabolismus lipidů genetika účinky léků MeSH
- nádorové buněčné linie účinky léků MeSH
- nádory slinivky břišní * farmakoterapie metabolismus MeSH
- proliferace buněk genetika účinky léků MeSH
- regulace genové exprese účinky léků MeSH
- simvastatin * farmakologie MeSH
- statiny * farmakologie MeSH
- Check Tag
- lidé MeSH
UBL5 protein, a structural homologue of ubiquitin, was shown to be involved in pre-mRNA splicing and transcription regulation in yeast and Caenorhabditis elegans, respectively. However, role of the UBL5 human orthologue is still elusive. In our study, we observed that endogenous human UBL5 that was localized in the nucleus, partially associates with Cajal bodies (CBs), nuclear domains where spliceosomal components are assembled. Simultaneous expression of exogenous UBL5 and coilin resulted in their nuclear colocalization in HeLa cells. The ability of UBL5 to interact with coilin was proved by GST pull-down assay using coilin that was either in vitro translated or extracted from HEK293T cells. Further, our results showed that the UBL5-coilin interaction was not influenced by coilin phosphorylation. These results suggest that UBL5 could be targeted to CBs via its interaction with coilin. Relation between human UBL5 protein and CBs is in the agreement with current observations about yeast orthologue Hub1 playing important role in alternative splicing.
- MeSH
- Cajalova tělíska metabolismus MeSH
- fluorescenční mikroskopie MeSH
- glutathiontransferasa genetika metabolismus MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- jaderné proteiny genetika metabolismus MeSH
- lidé MeSH
- oční proteiny genetika metabolismus MeSH
- rekombinantní fúzní proteiny genetika metabolismus MeSH
- transfekce MeSH
- ubikvitiny genetika metabolismus MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Small ubiquitin-related modifiers 1, 2 and 3 (SUMO-1, -2, -3), members of the ubiquitin-like protein family, can be conjugated to various cellular proteins. Conjugates of SUMO-2 and SUMO-3 (SUMO-2/3) accumulate in cells exposed to various stress stimuli or to MG132 treatment. Although the proteins modified by SUMO-2/3 during heat shock or under MG132 treatment have been identified, the significance of this modification remains unclear. Our data show that the inhibition of translation by puromycin or cycloheximide blocks both the heat shock and MG132 induced accumulation of SUMO-2/3 conjugates in HEK 293T and U2OS cells. However, the heat shock induced accumulation of SUMO-2/3 conjugates was restored by proteasome inhibition, which suggests that the inhibition of translation did not abolish SUMOylation itself. Furthermore, we show that some of the proteins truncated due to the treatment by low concentration of puromycin are SUMOylated in HEK 293T cells. We suggest that the SUMO-2/3 conjugates accumulating under the heat shock or MG132 treatment result largely from new protein synthesis and that portion of them is incorrectly folded.
- MeSH
- benzochinony farmakologie MeSH
- biologické modely MeSH
- cykloheximid farmakologie MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- inhibitory syntézy proteinů farmakologie MeSH
- leupeptiny farmakologie MeSH
- lidé MeSH
- makrocyklické laktamy farmakologie MeSH
- malé modifikační proteiny související s ubikvitinem metabolismus MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- proteosyntéza účinky léků MeSH
- puromycin farmakologie MeSH
- reakce na tepelný šok účinky léků MeSH
- sumoylace účinky léků MeSH
- ubikvitiny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Jednobuněčná eukaryota jako Giardia intestinalis nebo Entamoeba histolytica byly a jsou považovány za amitochondriální. Ovšem v roce 1999 byla u těchto druhů popsána nová organela - mitosom. Morfologicky je podobný mitochondriím, ale na druhou stranu neobsahuje vlastní DNA a není schopen tvořit energii ve formě ATP. Jakou má mitosom funkci? Jaký je původ mitosomu? Existuje příbuznost mezi mitochondrií a mitosomem? Odpověď na tyto otázky budou hledat následující řádky.
Unicellular eucaryotes (Giardia intestinalis, Entamoeba histolytica and next) were and still are considered to be amitochondrial. However, a new organelle, mitosome, was described in these species, in 1999. Morphologically, mitosome looks like mitochondria, but on the other hand mitosome has no intrinsic DNA and it is unable to produce energy in the form of ATP. What is function of mitosome? What is the origin of mitosome? Does any relationship exist between mitosome and mitochondria? This article searches the answers.
