Estrogen receptor alpha (ER) is a key biomarker for breast cancer, and the presence or absence of ER in breast and other hormone-dependent cancers decides treatment regimens and patient prognosis. ER is activated after ligand binding - typically by steroid. 2682 steroid compounds were used in a molecular docking study to identify novel ligands for ER and to predict compounds that may show anticancer activity. The effect of the most promising compounds was determined by a novel luciferase reporter assay. Two compounds, 7 and 12, showing ER inhibitory activity comparable to clinical inhibitors such as tamoxifen or fulvestrant were selected. We propose that the inhibitory effect of compounds 7 and 12 on ER is related to the presence of a double bond in their D-ring, which may protect against ER activation by reducing the electron density of the keto group, or may undergo metabolism leading to an active compound. Western blotting revealed that compound 12 decreased the level of ER in the breast cancer cell line MCF7, which was associated with reduced expression of both isoforms of the progesterone receptor, a well-known downstream target of ER. However, compound 12 has a different mechanism of action from fulvestrant. Furthermore, we found that compound 12 interferes with mitochondrial functions, probably by disrupting the electron transport chain, leading to induction of the intrinsic apoptotic pathway even in ER-negative breast cancer cells. In conclusion, the combination of computational and experimental methods shown here represents a rapid approach to determine the activity of compounds towards ER. Our data will not only contribute to research focused on the regulation of ER activity but may also be useful for the further development of novel steroid receptor-targeted drugs applicable in clinical practice.
- MeSH
- alfa receptor estrogenů genetika metabolismus MeSH
- estradiol farmakologie terapeutické užití MeSH
- estron * farmakologie MeSH
- fulvestrant farmakologie terapeutické užití MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory prsu * farmakoterapie metabolismus MeSH
- receptory pro estrogeny metabolismus MeSH
- simulace molekulového dockingu MeSH
- tamoxifen farmakologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The fast-track process to approve vaccines against COVID-19 has raised questions about their safety, especially in relation to fertility. Over the last 2 years, studies have appeared monitoring female fertility, especially from assisted reproduction centers or in animal experiments. However, studies monitoring healthy populations are still limited. The aim of our study was to monitor the relevant parameters of female fertility (sex and other steroids, LH, FSH, SHBG, Antimüllerian hormone and antral follicle count) before and then 2-4 months after the third dose of vaccination against COVID-19 in a group of 25 healthy fertile woman. In addition, anti-SARS-CoV-2 and anti-SARS-CoV-2S antibodies were determined. We did not observe significant changes in the measured parameters before and after the third dose of vaccination. By comparing levels of the analytes with antibodies indicating a prior COVID-19 infection, we found that women who had experienced the disease had statistically lower levels of estrone, estradiol, SHBG and 5α-dihydroprogesterone, and conversely, higher levels of androgen active dehydroepiandrosterone and dihydrotestosterone. Our results confirm that vaccination does not affect female fertility, and that what fertile women should be worried about is not vaccination, but rather COVID-19 infection itself.
- MeSH
- 20-alfa-dihydroprogesteron MeSH
- androgeny MeSH
- antimülleriánský hormon * MeSH
- COVID-19 * prevence a kontrola MeSH
- dehydroepiandrosteron MeSH
- dihydrotestosteron MeSH
- estradiol MeSH
- estron MeSH
- fertilita MeSH
- folikuly stimulující hormon MeSH
- lidé MeSH
- vakcíny proti COVID-19 MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
A development of robust and rapid method with simple sample preparation for the analysis of steroids of C18-, C19-, C21- families is of interest of many research groups. Here we present a novel LC-MS/MS method for the simultaneous quantification of 32 steroid hormones in human plasma. Twenty-two of them were analyzed directly without the need for derivatization, while ten were derivatized with 2-fluoro-1-methylpyridinium p-toluenesulfonate. The steroids were separated on a C18 column with a gradient elution consisting of methanol and water with the addition of 0.1% formic acid. The mass spectrometer was operated in positive ESI mode. Validation demonstrated that the method was applicable for the quantitative analysis of two C18- steroids (estrone, estradiol), nineteen C19- steroids (testosterone, epitestosterone, dihydrotestosterone, 11-ketodihydrotestosterone, 11β-hydroxyandrostenedione, 11β-hydroxytestosterone, 11-ketotestosterone, dehydroepiandrosterone, 7α-hydroxydehydroepiandrosterone, 7β-hydroxydehydroepiandrosterone, 7-ketodehydroepiandrosterone, androsterone, epiandrosterone, androstenedione, androstenediol, 5α-androstane-3α,17β-diol, 5α-androstane-3β,17β-diol, 5β-androstane-3α,17β-diol, 5β-androstane-3β,17β-diol), and eleven C21- steroids (cortisol, 21-deoxycortisol, 11-deoxycortisol, cortisone, corticosterone, 11-deoxycorticosterone, pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, 5α-dihydroprogesterone). The lower limits of quantification are appropriate for analyses in both physiological and various pathophysiological conditions. The accuracy, intra- and inter-day precision values as well as stability tests were in accordance with FDA Guidelines. The method will be a useful tool in investigating the mechanisms of steroid-related diseases and will serve as a steppingstone for the development of other methods for steroid analyses in various biological matrices such as prostate tissue, cerebrospinal fluid, urine, seminal fluid, and saliva.
- MeSH
- androgeny MeSH
- androstendion * MeSH
- chromatografie kapalinová metody MeSH
- estron MeSH
- lidé MeSH
- tandemová hmotnostní spektrometrie * metody MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
In comparison with analytical tools, bioassays provide higher sensitivity and more complex evaluation of environmental samples and are indispensable tools for monitoring increasing in anthropogenic pollution. Nevertheless, the disadvantage in cellular assays stems from the material variability used within the assays, and an interlaboratory adaptation does not usually lead to satisfactory test sensitivities. The aim of this study was to evaluate the influence of material variability on CXCL12 secretion by T47D cells, the outcome of the CXCL-test (estrogenic activity assay). For this purpose, the cell line sources, sera suppliers, experimental and seeding media, and the amount of cell/well were tested. The multivariable linear model (MLM), employed as an innovative approach in this field for parameter evaluation, identified that all the tested parameters had significant effects. Knowledge of the contributions of each parameter has permitted step-by-step optimization. The most beneficial approach was seeding 20,000 cells/well directly in treatment medium and using DMEM for the treatment. Great differences in both basal and maximal cytokine secretions among the three tested cell lines and different impacts of each serum were also observed. Altogether, both these biologically based and highly variable inputs were additionally assessed by MLM and a subsequent two-step evaluation, which revealed a lower variability and satisfactory reproducibility of the test. This analysis showed that not only parameter and procedure optimization but also the evaluation methodology must be considered from the perspective of interlaboratory method adaptation. This overall methodology could be applied to all bioanalytical methods for fast multiparameter and accurate analysis.
Many enzymes from the short-chain dehydrogenase/reductase superfamily (SDR) have already been well characterized, particularly those that participate in crucial biochemical reactions in the human body (e.g. 11β-hydroxysteroid dehydrogenase 1, 17β-hydroxysteroid dehydrogenase 1 or carbonyl reductase 1). Several other SDR enzymes are completely or almost completely uncharacterized, such as DHRS1 (also known as SDR19C1). Based on our in silico and experimental approaches, DHRS1 is described as a likely monotopic protein that interacts with the membrane of the endoplasmic reticulum. The highest expression level of DHRS1 protein was observed in human liver and adrenals. The recombinant form of DHRS1 was purified using the detergent n-dodecyl-β-D-maltoside, and DHRS1 was proven to be an NADPH-dependent reductase that is able to catalyse the in vitro reductive conversion of some steroids (estrone, androstene-3,17-dione and cortisone), as well as other endogenous substances and xenobiotics. The expression pattern and enzyme activities fit to a role in steroid and/or xenobiotic metabolism; however, more research is needed to fully clarify the exact biological function of DHRS1.
- MeSH
- dehydrogenasy/reduktasy s krátkým řetězcem metabolismus MeSH
- endoplazmatické retikulum metabolismus MeSH
- estron metabolismus MeSH
- HeLa buňky MeSH
- játra metabolismus MeSH
- kortison metabolismus MeSH
- lidé MeSH
- nadledviny metabolismus MeSH
- nádorové buněčné linie MeSH
- oxidoreduktasy genetika metabolismus MeSH
- rekombinantní proteiny biosyntéza genetika MeSH
- sekvence aminokyselin MeSH
- Sf9 buňky MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Compounds with estrogenic potencies and their adverse effects in surface waters have received much attention. Both anthropogenic and natural compounds contribute to overall estrogenic activity in freshwaters. Recently, estrogenic potencies were also found to be associated with cyanobacteria and their blooms in surface waters. The present study developed and compared the solid phase extraction and LC-MS/MS analytical approaches for determination of phytoestrogens (8 flavonoids - biochanin A, coumestrol, daidzein, equol, formononetin, genistein, naringenin, apigenin - and 5 sterols - ergosterol, β-sitosterol, stigmasterol, campesterol, brassicasterol) and cholesterol in water. The method was used for analyses of samples collected in stagnant water bodies dominated by different cyanobacterial species. Concentrations of individual flavonoids ranged from below the limit of detection to 3.58 ng/L. Sterols were present in higher amounts up to 2.25 μg/L. Biological potencies of these phytoestrogens in vitro were characterized using the hERα-HeLa-9903 cell line. The relative estrogenic potencies (compared to model estrogen - 17β-estradiol) of flavonoids ranged from 2.25E-05 to 1.26E-03 with coumestrol being the most potent. None of the sterols elicited estrogenic response in the used bioassay. Estrogenic activity was detected in collected field water samples (maximum effect corresponding to 2.07 ng/L of 17β-estradiol equivalents, transcriptional assay). At maximum phytoestrogens accounted for only 1.56 pg/L of 17β-estradiol equivalents, contributing maximally 8.5% of the total estrogenicity of the water samples. Other compounds therefore, most likely of anthropogenic origin such as steroid estrogens, are probably the major drivers of total estrogenic effects in these surface waters.
- MeSH
- chemické látky znečišťující vodu analýza MeSH
- cholestadienoly MeSH
- cholesterol analogy a deriváty MeSH
- estradiol analýza MeSH
- estrogeny analýza MeSH
- estron analýza MeSH
- fytoestrogeny analýza MeSH
- fytosteroly MeSH
- genistein analýza MeSH
- HeLa buňky MeSH
- isoflavony analýza MeSH
- lidé MeSH
- receptory pro estrogeny metabolismus MeSH
- sinice účinky léků metabolismus MeSH
- sitosteroly analýza MeSH
- sladká voda MeSH
- steroly analýza MeSH
- tandemová hmotnostní spektrometrie MeSH
- voda MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- dihydrotestosteron * krev MeSH
- estradiol * krev MeSH
- estron * krev MeSH
- folikuly stimulující hormon krev MeSH
- globulin vázající pohlavní hormony metabolismus MeSH
- lidé MeSH
- longitudinální studie MeSH
- luteinizační hormon krev MeSH
- míra přežití MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- stárnutí * krev MeSH
- testosteron * krev MeSH
- zdravotnické přehledy MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Publikační typ
- komentáře MeSH
- souhrny MeSH
- MeSH
- estetrol chemie MeSH
- estradiol * fyziologie chemie MeSH
- estriol * fyziologie chemie MeSH
- estron * fyziologie chemie MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- MeSH
- algoritmy MeSH
- androstendion * krev MeSH
- dehydroepiandrosteronsulfát * krev MeSH
- dospělí MeSH
- estradiol * krev MeSH
- estron * krev MeSH
- F faktor MeSH
- globulin vázající pohlavní hormony * analýza MeSH
- kohortové studie MeSH
- lidé MeSH
- polysomnografie MeSH
- poruchy iniciace a udržování spánku epidemiologie krev patofyziologie MeSH
- poruchy spánku a bdění epidemiologie krev patofyziologie MeSH
- poruchy spánku z vnitřních příčin * epidemiologie krev patofyziologie MeSH
- prevalence MeSH
- průřezové studie MeSH
- stupeň závažnosti nemoci MeSH
- syndromy spánkové apnoe epidemiologie krev patofyziologie MeSH
- testosteron * MeSH
- věkové faktory MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- komentáře MeSH
- souhrny MeSH
- Geografické názvy
- Německo MeSH
Zhoubné nádory ledvinného parenchymu tvoří u dospělých 1–2% všech zhoubných nádorů. Nejčastějším nádorem je karcinom (1). Většina karcinomů je sporadických, jen asi 4% jsou familiární. Smíšené nádory jsou vzácné a při jejich výskytu nesmíme opomenout, že to mohou být tzv. hereditární karcinomy (2).
Malignant tumours of the renal parenchyma account for 1%–2% of all malignant tumours in adults. Carcinoma is the most common tumour. Most carcinomas are sporadic, with only about 4% being familial. Mixed tumours are rare and, in the case of their occurrence, one must bear in mind that these can be so-called hereditary carcinomas.
- Klíčová slova
- hereditární karcinom, Knudsonova teorie dvou zásahů,
- MeSH
- Birtův-Hoggův-Dubeův syndrom diagnóza genetika ultrasonografie MeSH
- dědičné nádorové syndromy MeSH
- estron genetika MeSH
- genetické testování MeSH
- lidé MeSH
- nádory komplexní a smíšené MeSH
- nádory ledvin * genetika vrozené MeSH
- senioři MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- kazuistiky MeSH
