Nuclear factor of activated T cells (NFAT) as a new component of the signal transduction pathway in glioma cells.
Cyclosporin A (CsA) exerts its immunosuppressive effect by inhibiting the activity of nuclear factor of activated T cells (NFAT), thus preventing transcriptional induction of several cytokine genes. This effect is mediated through inactivation of the phosphatase calcineurin, which inhibits translocation of an NFAT component to the nucleus. We have previously reported that CsA inhibits the growth of rat C6 glioma cells in a dose-dependent manner and induces apoptotic cell death. Here, we report that NFAT DNA-binding activity is present in the nuclear extracts from C6 glioma cells and that CsA treatment inhibits the formation of a functional NFAT complex. We provide evidence for the presence of a group of NFATc proteins in proliferating glioma cells. Immunoblot analyses show that stimulation of C6 glioma cells with a calcium-inducing agent, ionomycin, alters NFATc migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This alteration is inhibited by simultaneous treatment with CsA, suggesting a calcineurin involvement in the regulation of glioma NFATc proteins. Direct immunofluorescence reveals the presence of NFATc proteins in nuclei of proliferating glioma cells and their disappearance in CsA-treated cells. These data point to a new mechanism of transcription regulation in glioma cells and provide an explanation for the observed sensitivity of glioma cells to CsA.
Mosieniak
G
G
Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, Warsaw, Poland.
Pyrzynska
B
B
Kaminska
B
B
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002273:Carcinogens / Q000494:pharmacology; D017362:Cyclic AMP Response Element-Binding Protein / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D005910:Glioma; D007376:Interleukin-2 / Q000235:genetics; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins / Q000502:physiology; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000502:physiology; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013601:T-Lymphocytes / Q000502:physiology; D018808:Transcription Factor AP-1 / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000502:physiology
D002273:Carcinogens; D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D007376:Interleukin-2; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D016572:Cyclosporine