118 research outputs found
Endothelin receptor B antagonists decrease glioma cell viability independently of their cognate receptor
Background:
Endothelin receptor antagonists inhibit the progression of many cancers, but research into their influence on glioma has been limited.
Methods:
We treated glioma cell lines, LN-229 and SW1088, and melanoma cell lines, A375 and WM35, with two endothelin receptor type B (ETRB)-specific antagonists, A-192621 and BQ788, and quantified viable cells by the capacity of their intracellular esterases to convert non-fluorescent calcein AM into green-fluorescent calcein. We assessed cell proliferation by labeling cells with carboxyfluorescein diacetate succinimidyl ester and quantifying the fluorescence by FACS analysis. We also examined the cell cycle status using BrdU/propidium iodide double staining and FACS analysis. We evaluated changes in gene expression by microarray analysis following treatment with A-192621 in glioma cells. We examined the role of ETRB by reducing its expression level using small interfering RNA (siRNA).
Results:
We report that two ETRB-specific antagonists, A-192621 and BQ788, reduce the number of viable cells in two glioma cell lines in a dose- and time-dependent manner. We describe similar results for two melanoma cell lines. The more potent of the two antagonists, A-192621, decreases the mean number of cell divisions at least in part by inducing a G2/M arrest and apoptosis. Microarray analysis of the effects of A-192621 treatment reveals up-regulation of several DNA damage-inducible genes. These results were confirmed by real-time RT-PCR. Importantly, reducing expression of ETRB with siRNAs does not abrogate the effects of either A-192621 or BQ788 in glioma or melanoma cells. Furthermore, BQ123, an endothelin receptor type A (ETRA)-specific antagonist, has no effect on cell viability in any of these cell lines, indicating that the ETRB-independent effects on cell viability exhibited by A-192621 and BQ788 are not a result of ETRA inhibition.
Conclusion:
While ETRB antagonists reduce the viability of glioma cells in vitro, it appears unlikely that this effect is mediated by ETRB inhibition or cross-reaction with ETRA. Instead, we present evidence that A-192621 affects glioma and melanoma viability by activating stress/DNA damage response pathways, which leads to cell cycle arrest and apoptosis. This is the first evidence linking ETRB antagonist treatment to enhanced expression of DNA damage-inducible genes
Metabolic regulation by p53
We are increasingly aware that cellular metabolism plays a vital role in diseases such as cancer, and that p53 is an important regulator of metabolic pathways. By transcriptional activation and other means, p53 is able to contribute to the regulation of glycolysis, oxidative phosphorylation, glutaminolysis, insulin sensitivity, nucleotide biosynthesis, mitochondrial integrity, fatty acid oxidation, antioxidant response, autophagy and mTOR signalling. The ability to positively and negatively regulate many of these pathways, combined with feedback signalling from these pathways to p53, demonstrates the reciprocal and flexible nature of the regulation, facilitating a diverse range of responses to metabolic stress. Intriguingly, metabolic stress triggers primarily an adaptive (rather than pro-apoptotic) p53 response, and p53 is emerging as an important regulator of metabolic homeostasis. A better understanding of how p53 coordinates metabolic adaptation will facilitate the identification of novel therapeutic targets and will also illuminate the wider role of p53 in human biology
mTOR: from growth signal integration to cancer, diabetes and ageing
In all eukaryotes, the target of rapamycin (TOR) signalling pathway couples energy
and nutrient abundance to the execution of cell growth and division, owing to the ability of TOR protein kinase to simultaneously sense energy, nutrients and stress and, in metazoans, growth factors. Mammalian TOR complex 1 (mTORC1) and mTORC2 exert their actions by regulating other important kinases, such as S6 kinase (S6K) and Akt. In the past few years, a significant advance in our understanding of the regulation and functions of mTOR has revealed the crucial involvement of this signalling pathway in the onset and progression of diabetes, cancer and ageing.National Institutes of Health (U.S.)Howard Hughes Medical InstituteWhitehead Institute for Biomedical ResearchJane Coffin Childs Memorial Fund for Medical Research (Postdoctoral Fellowship)Human Frontier Science Program (Strasbourg, France
Small molecule compounds targeting the p53 pathway: are we finally making progress?
Loss of function of p53, either through mutations in the gene or through mutations to other members of the pathway that inactivate wild-type p53, remains a critically important aspect of human cancer development. As such, p53 remains the most commonly mutated gene in human cancer. For these reasons, pharmacologic activation of the p53 pathway has been a highly sought after, yet unachieved goal in developmental therapeutics. Recently progress has been made not only in the discovery of small molecules that target wild-type and mutant p53, but also in the initiation and completion of the first in-human clinical trials for several of these drugs. Here, we review the current literature of drugs that target wild-type and mutant p53 with a focus on small-molecule type compounds. We discuss common means of drug discovery and group them according to their common mechanisms of action. Lastly, we review the current status of the various drugs in the development process and identify newer areas of p53 tumor biology that may prove therapeutically useful
Wolbachia Infections in Anopheles gambiae Cells: Transcriptomic Characterization of a Novel Host-Symbiont Interaction
The endosymbiotic bacterium Wolbachia is being investigated as a potential control agent in several important vector insect species. Recent studies have shown that Wolbachia can protect the insect host against a wide variety of pathogens, resulting in reduced transmission of parasites and viruses. It has been proposed that compromised vector competence of Wolbachia-infected insects is due to up-regulation of the host innate immune system or metabolic competition. Anopheles mosquitoes, which transmit human malaria parasites, have never been found to harbor Wolbachia in nature. While transient somatic infections can be established in Anopheles, no stable artificially-transinfected Anopheles line has been developed despite numerous attempts. However, cultured Anopheles cells can be stably infected with multiple Wolbachia strains such as wAlbB from Aedes albopictus, wRi from Drosophila simulans and wMelPop from Drosophila melanogaster. Infected cell lines provide an amenable system to investigate Wolbachia-Anopheles interactions in the absence of an infected mosquito strain. We used Affymetrix GeneChip microarrays to investigate the effect of wAlbB and wRi infection on the transcriptome of cultured Anopheles Sua5B cells, and for a subset of genes used quantitative PCR to validate results in somatically-infected Anopheles mosquitoes. Wolbachia infection had a dramatic strain-specific effect on gene expression in this cell line, with almost 700 genes in total regulated representing a diverse array of functional classes. Very strikingly, infection resulted in a significant down-regulation of many immune, stress and detoxification-related transcripts. This is in stark contrast to the induction of immune genes observed in other insect hosts. We also identified genes that may be potentially involved in Wolbachia-induced reproductive and pathogenic phenotypes. Somatically-infected mosquitoes had similar responses to cultured cells. The data show that Wolbachia has a profound and unique effect on Anopheles gene expression in cultured cells, and has important implications for mechanistic understanding of Wolbachia-induced phenotypes and potential novel strategies to control malaria
Model of subsystem software of ASCM RCB protection
Запропонована модель програмного забезпечення АСУ РХБ захисту на концептуальному та логічному рівнях подання та реалізована з використанням універсальної мови об’єктово-орієнтованого моделювання.Предложена модель программного обеспечения АСУ РХБ защиты на концептуальном и логическом уровнях представления.Offered model software to ASC of RCB at conceptual and logical levels of presentation
Experience in the Use of Scripting Languages in Distance Learning: Advantages and Disadvantages
Appearance of neutrino asymmetries in the process of expansion of the Universe, hierarchy of neutrino masses and CP violation
In this work, we study the appearance of neutrino asymmetries during the
expansion of the Universe. A mathematical model based on differential equations
was used to describe the processes of neutrino oscillations considering CP
violation and neutrino collisions. An analysis of the emergence of neutrino
asymmetry due to collisions of neutrinos with each other and due to CP
violation during neutrino oscillations is presented. It was discovered that the
asymmetry bifurcates into two states with positive and negative asymmetry for
the inverse hierarchy of neutrino masses. A state with a normal hierarchy of
neutrino masses is unstable and is not realized. It is shown that the maximum
CP violation is realized. The influence of this process on the dispersion of
the primordial mass fraction is calculated, which is confirmed by
astrophysical observations. Thus, the inverse hierarchy of neutrino masses is
realized. The CP violation in neutrino oscillations is maximum and the phase is
Comment: An error was discovered in the calculations when estimating the
collision frequency for various types of neutrinos. It turned out that the
same coefficient value was used in an uncontrolled manner, which is correct
only for electron neutrino
Vulvovaginal infections in the postantibiotic era. How to avoid aggression?
High incidence of recurring aerobic/nonspecific vaginitis is associated with the formation of stable pathological biofilm. The increasing antibiotic resistance does not allow to eliminate associations of conditionally pathogenic microorganisms and inhibits the growth of lactobacilli. Vaginal use of bacteriophages is a targeted antibiotic therapy. The study showed that multivalent bacteriophage (Sextaphage®) used as vaginal monotherapy is comparable with antibiotic in terms of treatment efficacy of aerobic vaginitis, does not reduce the level of lactobacilli and has a high compliance
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