Several papers have reported that calcium channel blocking drugs were associated with increased breast cancer risk and worsened prognosis. One of the most common signs of breast tumors is the presence of small deposits of calcium, known as microcalcifications. Therefore, we studied the effect of dihydropyridine nifedipine on selected calcium transport systems in MDA-MB-231 cells, originating from triple negative breast tumor and JIMT1 cells that represent a model of HER2-positive breast cancer, which possesses amplification of HER2 receptor, but cells do not response to HER2 inhibition treatment with trastuzumab. Also, we compared the effect of nifedipine on colorectal DLD1 and ovarian A2780 cancer cells. Both, inositol 1,4,5-trisphosphate receptor type 1 (IP3R1) and type 1 sodium calcium exchanger (NCX1) were upregulated due to nifedipine in DLD1 and A2780 cells, but not in breast cancer MDA-MB-231 and JIMT1 cells. On contrary to MDA-MB-231 and JIMT1 cells, in DLD1 and A2780 cells nifedipine induced apoptosis in a concentration-dependent manner. After NCX1 silencing and subsequent treatment with nifedipine, proliferation was decreased in MDA-MB-231, increased in DLD1 cells, and not changed in JIMT1 cells. Silencing of IP3R1 revealed increase in proliferation in DLD1 and JIMT1 cells, but caused decrease in proliferation in MDA-MB-231 cell line after nifedipine treatment. Interestingly, after nifedipine treatment migration was not significantly affected in any of tested cell lines after NCX1 silencing. Due to IP3R1 silencing, significant decrease in migration occurred in MDA-MB-231 cells after nifedipine treatment, but not in other tested cells. These results support different function of the NCX1 and IP3R1 in the invasiveness of various cancer cells due to nifedipine treatment.

• MeSH
• apoptóza účinky léků   genetika   MeSH
• blokátory kalciových kanálů farmakologie   MeSH
• inositol-1,4,5-trisfosfát - receptory genetika   metabolismus   MeSH
• kolorektální nádory genetika   metabolismus   patologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nádory prsu genetika   metabolismus   patologie   MeSH
• nádory vaječníků genetika   metabolismus   patologie   MeSH
• nifedipin farmakologie   MeSH
• pohyb buněk účinky léků   genetika   MeSH
• proliferace buněk účinky léků   genetika   MeSH
• protinádorové látky imunologicky aktivní farmakologie   MeSH
• pumpa pro výměnu sodíku a vápníku genetika   metabolismus   MeSH
• receptor erbB-2 genetika   metabolismus   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• RNA interference MeSH
• trastuzumab farmakologie   MeSH
• triple-negativní karcinom prsu genetika   metabolismus   patologie   MeSH
• vápníková signalizace účinky léků   genetika   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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
• antitumorózní látky farmakologie   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
• 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

Haloperidol is an antipsychotic agent that primarily acts as an antagonist of D2 dopamine receptors. Besides other receptor systems, it targets sigma 1 receptors (σ1Rs) and inositol 1,4,5-trisphosphate receptors (IP3Rs). Aim of this work was to investigate possible changes in IP3Rs and σ1Rs resulting from haloperidol treatment and to propose physiological consequences in differentiated NG-108 cells, i.e., effect on cellular plasticity. Haloperidol treatment resulted in up-regulation of both type 1 IP3Rs (IP3R1s) and σ1Rs at mRNA and protein levels. Haloperidol treatment did not alter expression of other types of IP3Rs. Calcium release from endoplasmic reticulum (ER) mediated by increased amount of IP3R1s elevated cytosolic calcium and generated ER stress. IP3R1s were bound to σ1Rs, and translocation of this complex from ER to nucleus occurred in the group of cells treated with haloperidol, which was followed by increased nuclear calcium levels. Haloperidol-induced changes in cytosolic, reticular, and nuclear calcium levels were similar when specific σ1 blocker -BD 1047- was used. Changes in calcium levels in nucleus, ER, and cytoplasm might be responsible for alterations in cellular plasticity, because length of neurites increased and number of neurites decreased in haloperidol-treated differentiated NG-108 cells.

• MeSH
• antipsychotika farmakologie   MeSH
• buněčná diferenciace účinky léků   fyziologie   MeSH
• haloperidol farmakologie   MeSH
• inositol-1,4,5-trisfosfát - receptory metabolismus   MeSH
• krysa rodu rattus MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• neuroplasticita účinky léků   fyziologie   MeSH
• receptory sigma metabolismus   MeSH
• vazba proteinů účinky léků   fyziologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Knowledge about the expression and thus a role of enzymes that produce endogenous H2S - cystathionine-β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase - in renal tumors is still controversial. In this study we aimed to determine the expression of these enzymes relatively to the expression in unaffected part of kidney from the same patient and to found relation of these changes to apoptosis. To evaluate patient's samples, microarray and immunohistochemistry was used. METHODS: To determine the physiological importance, we used RCC4 stable cell line derived from clear cell renal cell carcinoma, where apoptosis induction by a mixture of five chemotherapeutics with/without silencing of H2S-producing enzymes was detected. Immunofluorescence was used to determine each enzyme in the cells. RESULTS: In clear cell renal cell carcinomas, expression of H2S-producing enzymes was mostly decreased compared to a part of kidney that was distal from the tumor. To evaluate a potential role of H2S-producing enzymes in the apoptosis induction, we used RCC4 stable cell line. We have found that silencing of cystathionine-β-synthase and cystathionine γ-lyase prevented induction of apoptosis. Immunofluorescence staining clearly showed that these enzymes were upregulated during apoptosis in RCC4 cells. CONCLUSION: Based on these results we concluded that in clear cell renal cell carcinoma, reduced expression of the H2S-producing enzymes, mainly cystathionine γ-lyase, might contribute to a resistance to the induction of apoptosis. Increased production of the endogenous H2S, or donation from the external sources might be of a therapeutic importance in these tumors.

• MeSH
• apoptóza * MeSH
• cystathionin-beta-synthasa genetika   metabolismus   MeSH
• cystathionin-gama-lyasa genetika   metabolismus   MeSH
• dospělí MeSH
• karcinom z renálních buněk patologie   chirurgie   MeSH
• ledviny metabolismus   patologie   chirurgie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• malá interferující RNA metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory ledvin patologie   chirurgie   MeSH
• nefrektomie MeSH
• RNA interference MeSH
• senioři MeSH
• sulfan metabolismus   MeSH
• upregulace MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND/AIMS: Melatonin is a hormone transferring information about duration of darkness to the organism and is known to modulate several signaling pathways in the cells, e.g. generation of endoplasmic reticulum stress, oxidative status of the cells, etc. Melatonin has been shown to exert antiproliferative and cytotoxic effects on various human cancers. We proposed that this hormone can differently affect tumour cells and healthy cells. METHODS: We compared the effect of 24 h melatonin treatment on calcium transport (by fluorescent probes FLUO-3AM and Rhod-5N), ER stress (determined as changes in the expression of CHOP, XBP1 and fluorescently, using Thioflavin T), ROS formation (by CellROX® Green/Orange Reagent) and apoptosis induction (by Annexin-V-FLUOS/propidiumiodide) in two tumour cell lines - ovarian cancer cell line A2780 and stable cell line DLD1 derived from colorectal carcinoma, with non-tumour endothelial cell line EA.hy926. RESULTS: Melatonin increased apoptosis in both tumour cell lines more than twice, while in EA.hy926 cells the apoptosis was increased only by 30%. As determined by silencing with appropriate siRNAs, both, type 1 sodium/calcium exchanger and type 1 IP3 receptor are involved in the apoptosis induction. Antioxidant properties of melatonin were significantly increased in EA.hy926 cells, while in tumour cell lines this effect was much weaker. CONCLUSION: Taken together, melatonin has different antioxidative effects on tumour cells compared to non-tumour ones; it also differs in the ability to induce apoptosis through the type 1 sodium/calcium exchanger, and type 1 IP3 receptor. Different targeting of calcium transport systems in tumour and normal, non-tumour cells is suggested as a key mechanism how melatonin can exert its anticancer effects. Therefore, it might have a potential as a novel therapeutic implication in cancer treatment.

• MeSH
• apoptóza účinky léků   MeSH
• cytosol metabolismus   MeSH
• fluorescenční mikroskopie MeSH
• inositol-1,4,5-trisfosfát - receptory antagonisté a inhibitory   genetika   MeSH
• lidé MeSH
• malá interferující RNA metabolismus   MeSH
• melatonin toxicita   MeSH
• nádorové buněčné linie MeSH
• pumpa pro výměnu sodíku a vápníku antagonisté a inhibitory   genetika   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• RNA interference MeSH
• stres endoplazmatického retikula účinky léků   MeSH
• transkripční faktor CHOP genetika   metabolismus   MeSH
• vápník metabolismus   MeSH
• XBP1 genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

In this study we show that anti-tumor effect of sulforaphane (SFN) is partially realized through the type 1 inositol 1,4,5-trisphosphate receptor (IP3R1). This effect was verified in vitro on three different stable cell lines and also in vivo on the model of nude mice with developed tumors. Early response (6 hours) of A2780 ovarian carcinoma cells to SFN treatment involves generation of mitochondrial ROS and increased transcription of NRF2 and its downstream regulated genes including heme oxygenase 1, NAD(P)H:quinine oxidoreductase 1, and KLF9. Prolonged SFN treatment (24 hours) upregulated expression of NRF2 and IP3R1. SFN induces a time-dependent phosphorylation wave of HSP27. Use of IP3R inhibitor Xestospongin C (Xest) attenuates both SFN-induced apoptosis and the level of NRF2 protein expression. In addition, Xest partially attenuates anti-tumor effect of SFN in vivo. SFN-induced apoptosis is completely inhibited by silencing of IP3R1 gene but only partially blocked by silencing of NRF2; silencing of IP3R2 and IP3R3 had no effect on these cells. Xest inhibitor does not significantly modify SFN-induced increase in the rapid activity of ARE and AP1 responsive elements. We found that Xest effectively reverses the SFN-dependent increase of nuclear content and decrease of reticular calcium content. In addition, immunofluorescent staining with IP3R1 antibody revealed that SFN treatment induces translocation of IP3R1 to the nucleus. Our results clearly show that IP3R1 is involved in SFN-induced apoptosis through the depletion of reticular calcium and modulation of transcription factors through nuclear calcium up-regulation.

• MeSH
• aktivace transkripce účinky léků   MeSH
• antikarcinogenní látky farmakologie   terapeutické užití   MeSH
• antioxidační responzivní elementy MeSH
• apoptóza účinky léků   MeSH
• buněčné jádro metabolismus   MeSH
• endoplazmatické retikulum metabolismus   MeSH
• faktor 2 související s NF-E2 metabolismus   MeSH
• hemoxygenasa-1 metabolismus   MeSH
• inositol-1,4,5-trisfosfát - receptory antagonisté a inhibitory   metabolismus   MeSH
• isothiokyanatany farmakologie   terapeutické užití   MeSH
• lidé MeSH
• makrocyklické sloučeniny farmakologie   MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• myši nahé MeSH
• myši MeSH
• NAD(P)H dehydrogenasa (chinon) metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory vaječníků farmakoterapie   patologie   MeSH
• oxazoly farmakologie   MeSH
• oxidační stres účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• transkripční faktory Krüppel-like metabolismus   MeSH
• upregulace MeSH
• vápník metabolismus   MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• Publikační typ
• abstrakt z konference MeSH

We have studied the effect of chronic thyroid status alterations on the myosin heavy chain (MyHC) isoform composition (by SDS-PAGE) and on MyHC mRNA levels (by RT-PCR) in the fast extensor digitorum longus (EDL) muscle of 7-month-old inbred Lewis strain female rats and compared this with corresponding results of the previously studied slow soleus muscle. Our findings show that in the EDL muscle, all four types 1, 2a, 2x/d and 2b of MyHC mRNA transcripts and protein isoforms are present in euthyroid, hypothyroid and hyperthyroid rats, i.e. after chronic treatment with methimazole and T3, respectively. This is in contrast with the soleus, where only MyHC1 and 2a protein isoforms are expressed under similar conditions. Except for 2x/d MyHC mRNA transcripts in the EDL muscles, there was always significant difference between hypothyroid and hyperthyroid rats both at mRNA and protein levels. From our results we can conclude that extended alteration of the thyroid status leads to typical changes in the expression of MyHC mRNA transcripts and MyHC protein isoforms in the fast EDL and the slow soleus muscles. These changes correspond to those described after shorter periods of altered thyroid status. The characteristic phenotype differences between soleus and EDL muscles remain, however, preserved even after 7 months of thyroid hormone status alteration.

• MeSH
• elektroforéza v polyakrylamidovém gelu metody   využití   MeSH
• financování vládou MeSH
• hormony štítné žlázy fyziologie   metabolismus   škodlivé účinky   MeSH
• kosterní svaly enzymologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí metody   využití   MeSH
• potkani inbrední LEW anatomie a histologie   genetika   růst a vývoj   MeSH
• stanovení celkové genové exprese metody   využití   MeSH
• těžké řetězce myosinu genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

• MeSH
• finanční podpora výzkumu jako téma MeSH
• hormony štítné žlázy fyziologie   MeSH
• kosterní svaly metabolismus   MeSH
• krysa rodu rattus MeSH
• pumpa pro výměnu sodíku a vápníku genetika   MeSH
• regulace genové exprese MeSH
• ryanodinový receptor vápníkového kanálu genetika   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

• Publikační typ
• abstrakt z konference MeSH