Encapsulation is a well-established method of biomaterial protection, controlled release, and efficient delivery. Here we evaluated encapsulation of monoclonal antibody M75 directed to tumor biomarker carbonic anhydrase IX (CA IX) into alginate microbeads (SA-beads) or microcapsules made of sodium alginate, cellulose sulfate, and poly(methylene-co-guanidine) (PMCG). M75 antibody release was quantified using ELISA and its binding properties were assessed by immunodetection methods. SA-beads showed rapid M75 antibody release in the first hour, followed by steady release during the whole experiment of 7 days. In contrast, the M75 release from PMCG capsules was gradual, reaching the maximum concentration on the 7th day. The release was more efficient at pH 6.8 compared to pH 7.4. The released antibody could recognize CA IX, and target the CA IX-positive cells in 3D spheroids. In conclusion, SA-beads and PMCG microcapsules can be considered as promising antibody reservoirs for targeting of cancer cells.
- MeSH
- antigeny nádorové imunologie metabolismus MeSH
- antitumorózní látky aplikace a dávkování MeSH
- buněčné sféroidy metabolismus MeSH
- karboanhydrasa IX imunologie metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- lidé MeSH
- mikrosféry * MeSH
- monoklonální protilátky aplikace a dávkování imunologie farmakokinetika MeSH
- nádorové biomarkery imunologie metabolismus MeSH
- nádorové buňky kultivované MeSH
- nádory metabolismus patologie MeSH
- PEG-DMA hydrogel * MeSH
- systémy cílené aplikace léků metody MeSH
- uvolňování léčiv 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
Metastatic pheochromocytoma continues to be an incurable disease, and treatment with conventional cytotoxic chemotherapy offers limited efficacy. In the present study, we evaluated a novel topoisomerase I inhibitor, LMP-400, as a potential treatment for this devastating disease. We found a high expression of topoisomerase I in human metastatic pheochromocytoma, providing a basis for the evaluation of a topoisomerase 1 inhibitor as a therapeutic strategy. LMP-400 inhibited the cell growth of established mouse pheochromocytoma cell lines and primary human tumor tissue cultures. In a study performed in athymic female mice, LMP-400 demonstrated a significant inhibitory effect on tumor growth with two drug administration regimens. Furthermore, low doses of LMP-400 decreased the protein levels of hypoxia-inducible factor 1 (HIF-1α), one of a family of factors studied as potential metastatic drivers in these tumors. The HIF-1α decrease resulted in changes in the mRNA levels of HIF-1 transcriptional targets. In vitro, LMP-400 showed an increase in the growth-inhibitory effects in combination with other chemotherapeutic drugs that are currently used for the treatment of pheochromocytoma. We conclude that LMP-400 has promising antitumor activity in preclinical models of metastatic pheochromocytoma and its use should be considered in future clinical trials.
- MeSH
- antitumorózní látky farmakologie MeSH
- benzodioxoly aplikace a dávkování farmakologie MeSH
- buňky PC12 MeSH
- DNA-topoisomerasy I metabolismus MeSH
- faktor 1 indukovatelný hypoxií - podjednotka alfa genetika metabolismus MeSH
- feochromocytom farmakoterapie enzymologie patologie MeSH
- hypoxie buňky MeSH
- inhibitory topoisomerasy I aplikace a dávkování farmakologie MeSH
- isochinoliny aplikace a dávkování farmakologie MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- myši nahé MeSH
- nádorové buněčné linie MeSH
- nádorové buňky kultivované MeSH
- nádory jater farmakoterapie enzymologie sekundární MeSH
- nádory nadledvin farmakoterapie enzymologie patologie MeSH
- nádory plic farmakoterapie enzymologie sekundární MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proliferace buněk účinky léků MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- synergismus léků MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Intramural MeSH
The aim of the present work was to study the effect of various stressors (hypoxia, cold, immobilization) on the gene expression of sigma receptors in the left ventricles of rat heart. We have clearly shown that gene expression of sigma receptors is upregulated by strong stress stimuli, such as immobilization and/or hypoxia. Nevertheless, cold as a milder stressor has no effect on sigma receptor's mRNA levels. Signalling cascade of sigma receptors is dependent on IP(3) receptors, since silencing of both, type 1 and 2 IP(3) receptors resulted in decreased mRNA levels of sigma receptors. Physiological relevance of sigma receptors in the heart is not clear yet. Nevertheless, based on the already published data we can assume that sigma receptors might participate in contractile responses in cardiomyocytes.
- MeSH
- financování organizované MeSH
- hypoxie MeSH
- imobilizace MeSH
- inbrední kmeny myší MeSH
- inositol-1,4,5-trisfosfát - receptory metabolismus MeSH
- kardiomyocyty metabolismus patologie MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- messenger RNA analýza MeSH
- myši MeSH
- potkani inbrední SHR MeSH
- regulace genové exprese MeSH
- RNA interference MeSH
- srdeční komory metabolismus patofyziologie MeSH
- terapeutická hypotermie MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- HeLa buňky patologie MeSH
- myši nahé MeSH
- nádorové buněčné linie patologie MeSH
- nádory děložního čípku diagnóza genetika klasifikace MeSH
- proteiny S100 diagnostické užití genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- kyselý déšť MeSH
- látky znečišťující vodu dějiny MeSH
- monitorování životního prostředí dějiny MeSH
- potravní řetězec MeSH
- ryby MeSH
- stromy MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Geografické názvy
- Evropa MeSH
