ABSTRACT
The effects of Hedgehog signaling inhibitor (cyclopamine) and activator (Shh) on drug resistance of U251-MG human glioma cells and human astrocyte culture to cisplatin, temozolomide, and doxorubicin were studied. Cyclopamine and Shh modified the drug resistance of U251-MG cells but not of human astrocytes. Experiments with cyclopamine, Shh, and chemical drugs can contribute to detection of the mechanisms of signaling effects on the drug resistance processes, while the experimental data can serve as one of the criteria for choosing individual chemotherapy for patients.
Subject(s)
Antineoplastic Agents/pharmacology , Drug Resistance, Neoplasm/genetics , Gene Expression Regulation, Neoplastic , Hedgehog Proteins/genetics , Neuroglia/drug effects , Signal Transduction/genetics , Astrocytes/cytology , Astrocytes/drug effects , Astrocytes/metabolism , Cell Differentiation , Cell Line, Tumor , Cells, Cultured , Cisplatin/pharmacology , Dacarbazine/analogs & derivatives , Dacarbazine/pharmacology , Doxorubicin/pharmacology , Hedgehog Proteins/antagonists & inhibitors , Hedgehog Proteins/metabolism , Humans , Neuroglia/metabolism , Neuroglia/pathology , Peptides/pharmacology , Temozolomide , Veratrum Alkaloids/pharmacologyABSTRACT
The review summarizes current knowledge on the Hedgehog signaling pathway, its role in normal embryogenesis and/or initiation and progression of neuro-oncological diseases, especially of high-grade gliomas, the most malignant neuroepithelial tumors. The main proteins forming the Hedgehog signaling pathway include Shh, PTCH1, SMO, HHIP, SUFU and GLI1 isoforms. Effects of other signaling pathways on the family of transcription factors GLI and other proteins are described. The review summarizes modern data about the impact of the Hedgehog signaling pathway on proliferation, migration activity and invasiveness, and also on tumor neoangiogenesis and tumor cell chemoresistance. The role of the Hedgehog signaling pathway in origin of cancer stem cells and epithelial-mesenchymal transition is also analyzed. Some prospects for new anticancer drugs acting on components of the Hedgehog signaling pathway inhibitors are demonstrated.
Subject(s)
Central Nervous System Neoplasms/metabolism , Drug Resistance, Neoplasm , Glioma/metabolism , Hedgehog Proteins/metabolism , Animals , Cell Movement , Cell Proliferation , Central Nervous System Neoplasms/pathology , Glioma/pathology , Hedgehog Proteins/genetics , Humans , Signal Transduction , Transcription Factors/genetics , Transcription Factors/metabolism , Zinc Finger Protein GLI1ABSTRACT
To study demyelination and remyelination processes and their response to different drugs, a protocol for modeling multiple sclerosis using the copper chelator cuprizone was developed. Magnetic resonance imaging confirmed the presence of demyelination lesions on week 4 of 0.6% cuprizone-containing diet. Immunohistochemical staining with polyclonal antibodies to glial fibrillary acidic protein (pAb GFAP) confirmed the increase in the number of reactive astrocytes on week 4 of diet and during remyelination (week 2 after diet). Analysis of neurophysiological functions in mice with cuprizone-induced demyelination revealed motor and behavioral deficits. This model can be used as a tool for preclinical studies of the efficiency of multiple sclerosis diagnostic and therapy.
Subject(s)
Cuprizone/pharmacology , Demyelinating Diseases/chemically induced , Multiple Sclerosis/chemically induced , Myelin Sheath/metabolism , Nerve Tissue Proteins/metabolism , Animals , Brain/diagnostic imaging , Disease Models, Animal , Female , Glial Fibrillary Acidic Protein , Magnetic Resonance Imaging , Male , Mice , Mice, Inbred C57BL , Multiple Sclerosis/therapy , RadiographyABSTRACT
The results of NSE purification procedure, as well as hybridoma technology of anti NSE monoclonal antibodies synthesis are presented. The employment of this procedure yielded highly purified NSE preparation. The immunization of BALB/C mice with NSE preparation led to sensitization of the immunocompetent cells, which could form hybridomes, producing the anti-NSE monoclonal antibodies, after the confluence with myeloma cells Sp 2/0-Ag 14. The ELISA test-system for NSE analysis was developed on the basis of highly purified NSE preparation and monoclonal anti-NSE antibodies. This system was characterized by high specificity, accuracy and reliability. This system may be recommended for analysis of blood-brain barrier functions in the neurological and psychiatric diseases.
Subject(s)
Antibodies, Monoclonal/immunology , Immunoenzyme Techniques/methods , Phosphopyruvate Hydratase/immunology , Animals , Antibodies, Monoclonal/isolation & purification , Brain/enzymology , Humans , Hybridomas/immunology , Mice , Mice, Inbred BALB C , Phosphopyruvate Hydratase/isolation & purificationABSTRACT
Methods of GFAP purification and obtaining of hybridoma cells producing monoclonal anti-GFAP antibodies and properties of GFAP preparation were described. The immunobloting data on specificity of obtained monoclonal antibodies are presented. A new method of GFAP immunoenzyme assay based on GFAP preparation and anti-GFAP antibodies was elaborated. Standardization of the immunoenzyme system was shown in tests for specificity, accuracy, and reproducibility.
Subject(s)
Antibodies, Monoclonal/immunology , Glial Fibrillary Acidic Protein/analysis , Glial Fibrillary Acidic Protein/immunology , Immunoenzyme Techniques/methods , Animals , Antibodies, Monoclonal/isolation & purification , Blotting, Western , Humans , Hybridomas , Mice , Mice, Inbred BALB C , Molecular Weight , Reproducibility of Results , Sensitivity and SpecificitySubject(s)
Cell Adhesion Molecules, Neuronal/chemistry , Cell Adhesion Molecules, Neuronal/physiology , Humans , Leukocyte L1 Antigen Complex , Membrane Glycoproteins/chemistry , Membrane Glycoproteins/physiology , Myelin-Associated Glycoprotein/chemistry , Myelin-Associated Glycoprotein/physiology , Neoplasms/diagnosis , Neural Cell Adhesion Molecules/chemistry , Neural Cell Adhesion Molecules/physiology , Structure-Activity RelationshipABSTRACT
The effect of exogenous phosphocreatine on ischemic myocardium was studied in experimental infarction in rabbits and in total ischemia of pig heart tissue (in vitro). It is shown that single dose administration of phosphocreatine is followed by its rapid clearance from blood plasma (with a half lifetime of 4-6 min), but constantly high plasma concentration of phosphocreatine can be maintained by its intravenous infusion. When administered by this method into rabbits during experimental myocardial infarction, phosphocreatine reduces by 40% the size of the necrotic zone. Morphological electron microscopic studies using a lanthanum tracer method showed significant protection of the sarcolemma of cardiomyocytes in the perinecrotic zone by phosphocreatine. In vitro studies on the model of total ischemia also showed significant protection of cardiac sarcolemma from irreversible ischemic injury and reduction in the rate of high-energy phosphate depletion in the presence of phosphocreatine in the extracellular space. Additionally, it is demonstrated that creatine kinase released during myocardial infarction into the blood flow and exogenous phosphocreatine administered intravenously may significantly inhibit platelet aggregation by rapid removal of ADP, and thus potentially improve microcirculation during myocardial infarction.
Subject(s)
Coronary Disease/drug therapy , Myocardial Infarction/drug therapy , Phosphocreatine/therapeutic use , Platelet Aggregation , Sarcolemma/pathology , Adult , Animals , Coronary Disease/blood , Coronary Disease/pathology , Creatine Kinase/blood , Dogs , Humans , Kinetics , Microscopy, Electron , Middle Aged , Myocardial Infarction/blood , Myocardial Infarction/pathology , Myocardium/pathology , Phosphocreatine/blood , RabbitsABSTRACT
It is shown that a single intravenous injection of phosphocreatine to man or to experimental animals is followed by its rapid clearance from blood serum. This clearance is biphasic in nature with a half-life of 3-5 min at the first rapid stage and of 20-33 min at the next slower stage. To maintain the constant phosphocreatine concentration in blood, it is necessary to infuse it at a rate comparable to that of clearance. In particular, in blood serum of man, the phosphocreatine concentration can be kept at the level of 0.2 mM if it is injected at a rate of 60 mg/h per kg bw.
