ABSTRACT
HBL-100 breast epithelial cells were cultured with a blocker (N-ethylmaleimide) and protector (1,4-dithioerythritol) of SH groups. The study assessed changes in redox potential of glutathione and thioredoxin systems, intensity of oxidative modification of proteins, ROS production, and cell proliferation. The roles of thioredoxin system and protein oxidative modification in HBL-100 cell proliferation under redox status modulation were established. The role of carbonylated thioredoxin in arrest of the cell cycle in S-phase was demonstrated, which could be used for targeted therapy of the diseases accompanied by oxidative stress and disturbed redox status.
Subject(s)
Thioredoxins/metabolism , Cell Line , Cell Proliferation/drug effects , Dithioerythritol/pharmacology , Ethylmaleimide/pharmacology , Glutathione/metabolism , Humans , Oxidation-Reduction , Oxidative Stress/drug effects , Protein Processing, Post-Translational , Reactive Oxygen Species/metabolismABSTRACT
Activation of free radical oxidation in different cell types, including breast epithelial cells, may result in damage to macromolecules, in particular, proteins taking part in regulation of cell proliferation and apoptosis. The glutathione, glutaredoxin and thioredoxin systems play an essential role in maintaining intracellular redox homeostasis. Due to this fact, modulation of cellular redox status under the effect of an SH group inhibitor and an SH group protector may be used as a model for studying the role of redox proteins and glutathione in regulating cell proliferation in different pathological processes. In this study we have evaluated the state of the thioredoxin, glutaredoxin and glutathione systems as well as their role in regulating proliferation of HBL-100 breast epithelial cells under redox status modulation with N-ethylmaleimide (NEM) and 1,4-dithioerythriol (DTE). Modulating the redox status of breast epithelial cells under the effect of NEM and DTE influences the functional activity of glutathione-dependent enzymes, glutaredoxin, thioredoxin, and thioredoxin reductase through changes in the GSH and GSSG concentrations. In HBL-100 cells under redox-status modulation, we have found an increase in the number of cells in the S-phase of the cell cycle and a decrease in the number of cells in the G0/G1 and G2/Ð phases, as opposed to the values in the intact culture. The proposed model of proliferative activity of cells under redox status modulation may be used for development of new therapeutic approaches for treatment of diseases accompanied by oxidative stress generation.
Subject(s)
Dithioerythritol/pharmacology , Enzyme Inhibitors/pharmacology , Epithelial Cells/drug effects , Ethylmaleimide/pharmacology , Protective Agents/pharmacology , Catalase/metabolism , Cell Cycle/drug effects , Cell Line , Cell Proliferation/drug effects , Epithelial Cells/cytology , Epithelial Cells/metabolism , Ethylmaleimide/antagonists & inhibitors , Flow Cytometry , Glutaredoxins/metabolism , Glutathione/metabolism , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Humans , Mammary Glands, Human/cytology , Mammary Glands, Human/drug effects , Mammary Glands, Human/metabolism , Oxidation-Reduction , Oxidative Stress/drug effects , Thioredoxin-Disulfide Reductase/metabolism , Thioredoxins/metabolismABSTRACT
The phosphatidylinositol-4-phosphate-5-kinase type IIa (PIP5K2A) gene has been proposed as a putative susceptibility gene for schizophrenia on both positional and functional grounds. The association between the (N251S)-PIP5K2A (rs10828317) variant and schizophrenia was found in German and Dutch populations but was not replicated in several other populations. The purpose of the study was to examine whether the previously implicated (N251S)-PIP5K2A variant influences susceptibility to schizophrenia in the Russian population of Siberia. Authors studied 355 patients with schizophrenic disorders from the Russian population of Siberia. The control group consisted of 100 healthy. Results confirm the association of the (N251S)-PIP5K2A (rs10828317) polymorphism with schizophrenia (p=0.04, OR=2.48, 95%CI=1.19--5.17 for the CC genotype). The association can be explained by the inability of mutant kinase to activate the phosphatidylinositol-4,5-biphosphate dependent proteins, including neuronal KCNQ channels and glutamate EAAT3 transporters, which leads to the lack of dopaminergic and glutamatergic control in schizophrenic patients carriers of this mutation.
Subject(s)
DNA/genetics , Ethnicity/genetics , Genetic Predisposition to Disease , Phosphotransferases (Alcohol Group Acceptor)/genetics , Polymorphism, Genetic , Schizophrenia/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Female , Gene Frequency , Genotype , Humans , Male , Middle Aged , Phosphotransferases (Alcohol Group Acceptor)/metabolism , Polymerase Chain Reaction , Prevalence , Schizophrenia/ethnology , Schizophrenia/metabolism , Siberia/epidemiology , Young AdultABSTRACT
Dyskinesias are involuntary muscle movements that occur spontaneously in Huntington's disease (HD) and after long-term treatments for Parkinson's disease (levodopa-induced dyskinesia; LID) or for schizophrenia (tardive dyskinesia, TD). Previous studies suggested that dyskinesias in these three conditions originate from different neuronal pathways that converge on overstimulation of the motor cortex. We hypothesized that the same variants of the N-methyl-D-aspartate receptor gene that were previously associated with the age of dyskinesia onset in HD were also associated with the vulnerability for TD and not LID. Genotyping patients with LID and TD revealed, however, that these two variants were dose-dependently associated with susceptibility to LID, but not TD. This suggested that LID, TD and HD might arise from the same neuronal pathways, but TD results from a different mechanism.
