Abstract
Members of the nuclear factor of activated T cells (NFAT) family have been identified as regulators of oncogenic transformation in several human malignancies. A prominent member of this family, NFAT1, is associated with tumor cell survival, apoptosis, migration and invasion. Here, we investigated the role of NFAT1 in glioma cells. In 111 clinical samples, microarray analysis demonstrated that NFAT1 was over-expressed in glioblastoma multiforme (GBM), compared with low-grade gliomas, a result confirmed by RT-PCR in 24 clinical samples and in the U87 and U251 cell lines. Immunohistochemistry and immunofluorescence stain indicated that over-expressed NFAT1 was mainly located in the nucleus, where it acted as a transcription factor. After treatment with the NFAT antagonist cyclosporin A (CsA) and FK506, levels of NFAT1 in the nuclei of U87 GBM cells were dramatically reduced. The invasive potential of U87 cells was reduced by the same treatment, as well as by inhibition of NFAT1 expression using small hairpin RNA. Proliferation of U87 cells was unaffected by CsA, FK506 and NFAT1 shRNA transfection. Clustering analysis and Pearson correlation analysis of microarray data showed that the expression of NFAT1 correlated with the expression of the invasion-related genes cyclooxygenase-2 (COX-2), matrix metalloproteinase-7 (MMP-7) and MMP-9, a result confirmed by in vitro analysis. These findings demonstrate that NFAT1 contributes to the invasive potential but not the proliferation of GBM cells, and suggest that CsA may find application as an adjuvant in combined treatment strategies for GBM.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Biopsy
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Brain Neoplasms / genetics*
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Brain Neoplasms / metabolism
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Brain Neoplasms / pathology
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Cell Cycle
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Cell Line, Tumor
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Cell Nucleus / drug effects
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Cell Nucleus / genetics*
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Cell Nucleus / metabolism
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Cell Nucleus / pathology
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Cell Proliferation
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Cyclooxygenase 2 / genetics
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Cyclooxygenase 2 / metabolism
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Cyclosporine / pharmacology
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Gene Expression Profiling
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Gene Expression Regulation, Neoplastic*
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Glioblastoma / genetics*
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Glioblastoma / metabolism
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Glioblastoma / pathology
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Humans
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Matrix Metalloproteinase 7 / genetics
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Matrix Metalloproteinase 7 / metabolism
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Matrix Metalloproteinase 9 / genetics
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Matrix Metalloproteinase 9 / metabolism
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NFATC Transcription Factors / antagonists & inhibitors
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NFATC Transcription Factors / genetics*
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NFATC Transcription Factors / metabolism
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Neoplasm Invasiveness
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Oligonucleotide Array Sequence Analysis
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Tacrolimus / pharmacology
Substances
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NFATC Transcription Factors
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NFATC2 protein, human
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RNA, Small Interfering
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Cyclosporine
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Cyclooxygenase 2
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PTGS2 protein, human
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MMP7 protein, human
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Matrix Metalloproteinase 7
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Matrix Metalloproteinase 9
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Tacrolimus
Grants and funding
This work was supported by grants from the National High Technology Research and Development Program of China (863) (No.2012AA02A508), National Natural Science Foundation of China (No.81172409) and Science and Technology Department of Liaoning Province (No.2011225034). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.