Cyanobacterial blooms are known sources of environmentally-occurring retinoid compounds, including all-trans and 9-cis retinoic acids (RAs). The developmental hazard for aquatic organisms has been described, while the implications for human health hazard assessment are not yet sufficiently characterized. Here, we employ a human neural stem cell model that can differentiate in vitro into a mixed culture of neurons and glia. Cells were exposed to non-cytotoxic 8-1000 nM all-trans or 9-cis RA for 9-18 days (DIV13 and DIV22, respectively). Impact on biomarkers was analyzed on gene expression (RT-qPCR) and protein level (western blot and proteomics) at both time points; network patterning (immunofluorescence) on DIV22. RA exposure significantly concentration-dependently increased gene expression of retinoic acid receptors and the metabolizing enzyme CYP26A1, confirming the chemical-specific response of the model. Expression of thyroid hormone signaling-related genes remained mostly unchanged. Markers of neural progenitors/stem cells (PAX6, SOX1, SOX2, NESTIN) were decreased with increasing RA concentrations, though a basal population remained. Neural markers (DCX, TUJ1, MAP2, NeuN, SYP) remained unchanged or were decreased at high concentrations (200-1000 nM). Conversely, (astro-)glial marker S100β was increased concentration-dependently on DIV22. Together, the biomarker analysis indicates an RA-dependent promotion of glial cell fates over neural differentiation, despite the increased abundance of neural protein biomarkers during differentiation. Interestingly, RA exposure induced substantial changes to the cell culture morphology: while low concentrations resulted in a network-like differentiation pattern, high concentrations (200-1000 nM RA) almost completely prevented such network patterning. After functional confirmation for implications in network function, such morphological features could present a proxy for network formation assessment, an apical key event in (neuro-)developmental Adverse Outcome Pathways. The described application of a human in vitro model for (developmental) neurotoxicity to emerging environmentally-relevant retinoids contributes to the evidence-base for the use of differentiating human in vitro models for human health hazard and risk assessment.
- MeSH
- alitretinoin * toxicita MeSH
- buněčná diferenciace MeSH
- lidé MeSH
- nervové kmenové buňky * účinky léků MeSH
- receptory kyseliny retinové genetika metabolismus MeSH
- retinoidy farmakologie MeSH
- tretinoin * toxicita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cyanobacteria are known for their ability to produce and release mixtures of up to thousands of compounds into the environment. Recently, the production of novel metabolites, retinoids, was reported for some cyanobacterial species along with teratogenic effects of samples containing these compounds. Retinoids are natural endogenous substances derived from vitamin A that play a crucial role in early vertebrate development. Disruption of retinoid signalling- especially during the early development of the nervous system- might lead to major malfunctions and malformations. In this study, the toxicity of cyanobacterial biomass samples from the field containing retinoids was characterized by in vivo and in vitro bioassays with a focus on the potential hazards towards nervous system development and function. Additionally, in order to identify the compounds responsible for the observed in vitro and in vivo effects the complex cyanobacterial extracts were fractionated (C18 column, water-methanol gradient) and the twelve obtained fractions were tested in bioassays. In all bioassays, all-trans retinoic acid (ATRA) was tested along with the environmental samples as a positive control. Retinoid-like activity (mediated via the retinoic acid receptor, RAR) was measured in the transgenic cell line p19/A15. The in vitro assay showed retinoid-like activity by specific interaction with RAR for the biomass samples. Neurotoxic effects of selected samples were studied on zebrafish (Danio rerio) embryos using the light/dark transition test (Viewpoint, ZebraLab system) with 120 hpf larvae. In the behavioural assay, the cyanobacterial extracts caused significant hyperactivity in zebrafish at 120 hpf after acute exposure (3 h prior to the measurement) at concentrations below the teratogenicity LOEC (0.2 g dw L-1). Similar effect was observed after exposure to fractions of the extracts with detected retinoid-like activity and additive effect was observed after combining the fractions. However, the effect on behaviour was not observed after exposure to ATRA only. To provide additional insight into the behavioural effects and describe the underlying mechanism gene expression of selected biomarkers was measured. We evaluated an array of 28 genes related to general toxicity, neurodevelopment, retinoid and thyroid signalling. We detected several affected genes, most notably, the Cyp26 enzymes that control endogenous ATRA concentration, which documents an effect on retinoid signalling.
- MeSH
- biomasa MeSH
- biotest MeSH
- chemické látky znečišťující vodu metabolismus toxicita MeSH
- chování zvířat účinky léků MeSH
- dánio pruhované růst a vývoj metabolismus MeSH
- embryo nesavčí účinky léků metabolismus MeSH
- exprese genu účinky léků MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- receptory kyseliny retinové genetika metabolismus MeSH
- sinice růst a vývoj metabolismus MeSH
- tretinoin metabolismus toxicita MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Background MDA5 and RIG-1 are the important intracellular receptors which detect microbial associated molecular patterns. Septicemia is a condition in which infection enters the bloodstream of the patients. The main intracellular mechanisms against septicemia are yet to be clarified. Therefore, this research study was aimed to evaluate expression of MDA5 and RIG-1 in the patients suffering from septicemia in comparison to healthy controls. Methods MDA5 and RIG-1 expression levels in 40 patients suffering from septicemia and 40 healthy controls were evaluated using Real-Time PCR technique. The sources of bacteria in the bloodstream of the patients suffering from septicemia were determined using microbial cultures. Results The results showed that mRNA levels of MDA5 and RIG-1 were significantly increased in the patients when compared to healthy controls. The results also revealed that the patients were infected with four bacteria including Escherichia coli, Staphylococcus aureus, Acinetobacter baumannii and Pseudomonas aeruginosa. mRNA levels of MDA5 and RIG-1 did not differ among patients with various bacterial infections. Conclusion Based on the results it seems that MDA5 and RIG-1 are the main intracellular immunity against the bacteria during septicemia and could be considered for their roles in induction of immunity.
- MeSH
- biologické markery MeSH
- imunita genetika imunologie MeSH
- krev imunologie mikrobiologie MeSH
- lidé MeSH
- melatoninové receptory genetika imunologie krev MeSH
- receptory kyseliny retinové genetika imunologie krev MeSH
- sepse * genetika imunologie mikrobiologie MeSH
- transkripční faktory MeSH
- Check Tag
- lidé MeSH
Sox3/SOX3 gene is considered to be one of the earliest neural markers in vertebrates. Despite the mounting evidence that Sox3/SOX3 is one of the key players in the development of the nervous system, limited data are available regarding the transcriptional regulation of its expression. This review is focused on the retinoic acid induced regulation of SOX3 gene expression, with particular emphasis on the involvement of retinoid receptors. Experiments with human embryonal carcinoma cells identified two response elements involved in retinoic acid/retinoid X receptor-dependent activation of the SOX3 gene expression: distal atypical retinoic acid-response element, consisting of two unique G-rich boxes separated by 49 bp, and proximal element comprising DR-3-like motif, composed of two imperfect hexameric half-sites. Importantly, the retinoic acid-induced SOX3 gene expression could be significantly down-regulated by a synthetic antagonist of retinoid receptors. This cell model provides a solid base for further studies on mechanism(s) underlying regulation of expression of SOX3 gene, which could improve the understanding of molecular signals that induce neurogenesis in the stem/progenitor cells both during development and in adulthood.
- MeSH
- buněčná diferenciace genetika účinky léků MeSH
- financování organizované MeSH
- genetické techniky využití MeSH
- kmenové buňky embryonálního karcinomu MeSH
- lidé MeSH
- metaanalýza jako téma MeSH
- modely genetické MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- neurogeneze genetika účinky léků MeSH
- receptory kyseliny retinové genetika MeSH
- regulace genové exprese fyziologie genetika účinky léků MeSH
- transkripční faktory SOXB1 genetika MeSH
- tretinoin MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- Publikační typ
- přehledy MeSH
Acute promyelocytic leukemia (APL) with atypical breakpoints in the promyelocytic leukemia (PML) and retinoic acid receptor-alpha (RARA) genes represents a rare leukemic event, which occurs preferentially in patients with variant types of the PML/RARA fusion gene. Here we report on a patient with APL with a unique PML/RARA fusion transcript that harbors a short type of this fusion gene, exhibiting unexpected results of standard PCR diagnostics. The detected transcript originates from fusion of PML exon 4 and a truncated form of transcription variant 2 of the RARA gene, with an additional 9 bp insertion. According to our knowledge, this differs from all previously described fusion transcripts.
- MeSH
- elektroforéza v agarovém gelu MeSH
- fúzní onkogenní proteiny genetika MeSH
- genetická transkripce MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- molekulární sekvence - údaje MeSH
- protein - isoformy genetika MeSH
- receptory kyseliny retinové genetika MeSH
- sekvence nukleotidů MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- práce podpořená grantem MeSH
Reactivation of tumour suppressor genes that have been silenced by promoter methylation is a very attractive molecular target for cancer therapy. The treatment of a squamous cervical cancer cell line, SiHa, with 20 µM curcumin and genistein resulted in demethylation of promoter of the RARß2 gene and led to the reactivation of the gene. The degree of methylation as observed by MSP decreased as the time period of treatment was increased from 72 h to 6 days. In HeLa cells (an adenocarcinoma cervical cancer cell line) there was also reversal of hypermethylation of the RARß2 gene after six days of treatment with 20 µM curcumin. However, allyl sulphide treatment (20 µM) did not cause the reversal of hypermethylation until 72 h of treatment in the SiHa cell line. This is the first report to show the reversal of hypermethylation of the RARß2 gene by genistein and curcumin in cervical cancer cell lines. Furthermore, these compounds acted as doublepronged agents as they caused apoptosis in the treated cervical cancer cell lines in addition to reversal of promoter hypermethylation.
- MeSH
- alylové sloučeniny farmakologie MeSH
- antitumorózní látky farmakologie terapeutické užití MeSH
- biologické přípravky farmakologie MeSH
- financování organizované MeSH
- genistein farmakokinetika terapeutické užití MeSH
- kurkumin farmakologie terapeutické užití MeSH
- lidé MeSH
- metylace DNA účinky léků MeSH
- nádorové buněčné linie MeSH
- nádory děložního čípku farmakoterapie genetika patofyziologie MeSH
- receptory kyseliny retinové genetika MeSH
- sulfidy farmakologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
Cellular signaling by glucocorticoid receptor and aryl hydrocarbon receptor is restricted by microtubules interfering agents (MIAs). This leads to down-regulation of drug metabolizing enzymes and drug interactions. Here we investigated the effects of all-trans-retinoic acid (ATRA) and MIAs, i.e. colchicine, nocodazole and taxol on the regulation of retinoic acid receptor (RAR) genes in primary cultures of rat hepatocytes. ATRA (1microM) down-regulated RARalpha and RARgamma mRNAs (decrease 23% and 41%, respectively) whereas it up-regulated RARbeta mRNA (4.3-fold induction). All MIAs diminished the expression of RARs in dose-dependent manner; the potency of MIAs increased in order NOC
- MeSH
- financování organizované MeSH
- genetická transkripce účinky záření MeSH
- HeLa buňky cytologie metabolismus účinky léků MeSH
- inhibitory proteasomu MeSH
- katalýza účinky léků MeSH
- kolchicin farmakologie MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- leupeptiny farmakologie MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- mikrotubuly účinky léků MeSH
- modulátory tubulinu farmakologie MeSH
- proteasomový endopeptidasový komplex MeSH
- receptory kyseliny retinové genetika metabolismus MeSH
- regulace genové exprese účinky záření MeSH
- termodynamika MeSH
- transglutaminasy metabolismus MeSH
- tretinoin farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
The rat model of N-methyl-N-nitrosourea (MNU)-induced mammary carcinomas is well-established animal model for breast cancer. This study was carried out to investigate whether hypothyroid (thyroidectomy or PTU treatment) or hyperthyroid status of female rats would affect MNU-induced mammary carcinogenesis with specific focus on both retinoid and rexinoid receptor expression in mammary tumours. Application of PTU before and during MNU-induced mammary gland carcinogenesis yielded in a marked decrease of the number and volume of tumours per animal, however, there was no effect of hypothyroid state in thyroidectomized rats as well as hyperthyroid state concerning the number and volume of tumours. Mammary tumours of in euthyroid group of MNU animals showed that there was no tumour, in which all of subtypes of retinoid and rexinoid receptors were expressed. A different pattern of expression of retinoid or rexinoid receptors was found either in MNU-induced mammary carcinomas in both hypothyroid and hyperthyroid rats.
- MeSH
- experimentální nádory mléčných žláz genetika chemicky indukované metabolismus MeSH
- exprese genu MeSH
- financování organizované MeSH
- hormony štítné žlázy fyziologie MeSH
- hypertyreóza MeSH
- hypotyreóza chemicky indukované MeSH
- karcinogeny MeSH
- krysa rodu rattus MeSH
- messenger RNA MeSH
- methylnitrosomočovina MeSH
- potkani Sprague-Dawley MeSH
- propylthiouracil MeSH
- receptory kyseliny retinové genetika metabolismus MeSH
- tyreoidektomie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- ženské pohlaví MeSH
- zvířata MeSH
