ABSTRACT
The aim of our study was to investigate the effect of recombinant human cytokine EMAP II (endothelial monocyte-activating polypeptide II) on the expression of MGMT gene, encoding repair enzyme O6-methylguanine-DNA methyltransferase (MGMT) in human cell cultures. The influence of EMAP II on cell proliferation was performed using routine MTT assay. Identification of MGMT in cell extracts was performed using Western blot analysis. We used cell lines: A102 (fibroblasts), CB-1 (umbilical cord blood stromal cells), 4BL6 (cells derived from peripheral blood). It was shown that cytokine EMAP II caused induction of MGMT expression in studied human cell lines. There was a decrease in cell number at high concentrations of this cytokine. It was found that the presence of cytokine EMAP II in serum-free growth medium leads to increasing of repair enzyme MGMT expression level in human cells in vitro.
Subject(s)
Antineoplastic Agents, Alkylating/adverse effects , Cell Survival/drug effects , Cytokines , DNA Repair/drug effects , Gene Expression/drug effects , Neoplasm Proteins , O(6)-Methylguanine-DNA Methyltransferase/metabolism , RNA-Binding Proteins , Antineoplastic Agents, Alkylating/administration & dosage , Blotting, Western , Cell Line, Tumor , Cell Proliferation/drug effects , Culture Media, Serum-Free , Cytokines/pharmacology , Cytokines/therapeutic use , DNA Damage/drug effects , Enzyme Induction/drug effects , Fetal Blood/cytology , Fibroblasts/cytology , Fibroblasts/drug effects , Fibroblasts/metabolism , Guanine/analogs & derivatives , Guanine/metabolism , Hepatocytes/cytology , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Methylation , Neoplasm Proteins/pharmacology , Neoplasm Proteins/therapeutic use , Neoplasms/drug therapy , Neoplasms/pathology , O(6)-Methylguanine-DNA Methyltransferase/genetics , RNA-Binding Proteins/pharmacology , RNA-Binding Proteins/therapeutic use , Stromal Cells/cytology , Stromal Cells/drug effects , Stromal Cells/metabolismABSTRACT
The influence of plant lectins on primary DNA damage repair and on expression of repair enzyme alkylguanine-DNA alkyltransferase (MGMT) in mammalian cells in vitro has been investigated. Those lectins have been shown to modify DNA damage repair process, and to induce the MGMT expression increasing its level in the used model system.
Subject(s)
DNA Damage , DNA Repair , Mesenchymal Stem Cells/drug effects , O(6)-Methylguanine-DNA Methyltransferase/biosynthesis , Plant Lectins/pharmacology , Animals , Blotting, Western , Cell Culture Techniques , Cell Line, Tumor , Cricetinae , Cricetulus , Enzyme Induction , Humans , Mesenchymal Stem Cells/enzymology , Mesenchymal Stem Cells/metabolismABSTRACT
Formation of single- and double strand breaks in DNA which may be discovered by microelectrophoresis in agarose gel is one of the criterion of genetical lesions in cells as a result of apoptosis or of genotoxic agent action. Genotoxic action of nickel chloride at the level of DNA of the individual cells in the initial culture of human embryonic haemopoietic cells was studied. It has been shown that about 2% cells in the studied in vitro populations were in the apoptosis state. Nickel chloride induced increasing of the frequency of formation of electrophoretic tracks of "comet" type with destroyed DNA.
