NFAT1-Mediated Regulation of NDEL1 Promotes Growth and Invasion of Glioma Stem-like Cells

Abstract

Glioma stem-like cells (GSC) promote tumor generation and progression. However, the mechanism of GSC induction or maintenance is largely unknown. We previously demonstrated that the calcium-responsive transcription factor nuclear factor of activated T cells-1 (NFAT1) is activated in glioblastomas and regulates the invasion of tumor cells. In this study, we further explored the role of NFAT1 in GSC. We found that NFAT1 expression was associated with an aggressive phenotype and predicted poor survival in gliomas. Compared with normal glioma cells, NFAT1 was upregulated in GSC. NFAT1 knockdown reduced GSC viability, invasion, and self-renewal in vitro and inhibited tumorigenesis in vivo, whereas NFAT1 overexpression enhanced the growth and invasion of GSCs. RNA sequencing showed that NFAT1 depletion was associated with reduced neurodevelopment protein 1-like 1 (NDEL1, a potential downstream target of NFAT1) expression, whereas NFAT1 overexpression induced NDEL1 expression. In addition, NFAT1 regulated the promoter activities of NDEL1, whereas rescue of NDEL1 in NFAT1-silenced GSC partially restored tumor growth and invasion. Upregulation of NFAT1-NDEL1 signaling elevated Erk activation, increased protein levels of stemness markers in GSC, and resulted in dedifferentiation of normal neuronal cells and astrocytes. Our results indicate that NFAT1 controls the growth and invasion of GSC partially through regulation of NDEL1. Targeting the NFAT1-NDEL1 axis therefore might be of potential benefit in the treatment of patients with glioma.Significance: NFAT1 controls the growth and invasion of GSCs, partially by regulating NDEL1. Targeting the NFAT1-NDEL1 axis might provide opportunities in treating patients with glioma. Immunofluorescence staining of NFAT1 in LN229, G10, and T98G GSCs. DAPI was used for nuclear staining. Scale bar, 25 mm. Results are presented as the mean AE SEM of triplicate samples from three independent experiments. Ã , P < 0.05; ÃÃ , P < 0.01.

MiR-18a-downregulated RORA inhibits the proliferation and tumorigenesis of glioma using the TNF-α-mediated NF-κB signaling pathway

“…The slices were then stained with an immunohistochemical labeling kit (MaxVision Biotechnology, Fuzhou, China) and imaged under a light microscope (Olympus, Tokyo, Japan). The German immunohistochemical score (30) was used to evaluate the staining intensity and expression levels.…”

“…The glioma cells and GSCs treated under different conditions were seeded into 96-well plates at 1 £ 10 3 cells/well and cultured for 5 days. Cell viability was detected using the CellTiter 96 Ò Aqueous Non-Radioactive Cell Proliferation Assay Kit (Promega, Madison, WI, USA) according to the manufacturer's instructions [30].…”

“…The neurosphere formation assay was performed as previously described [30]. Briefly, GSCs were seeded into 24-well plates at a density of 200 cells/well and cultured in fresh medium for 7 days.…”

“…Luciferase reporter assays were performed as previously described [29,30]. Briefly, the TNF-a reporter plasmids were constructed by Gene-Chem (Shanghai, China).…”

Autophagic Network Analysis of the Dual Effect of Sevoflurane on Neurons Associated with GABARAPL1 and 2

“…Proteins enriched in the top 10 pathways interacted with DPTs of sevoflurane from DTome. Because the GABAergic synapse [28], cAMP signaling pathways [29], and calcium signaling [30] are strongly related to neurotoxicity and neurodevelopment, the genes included in these signaling pathways were selected to investigate DPT-associated genes. Interestingly, bioinformatics analysis of the candidate genes revealed that GABARAPL1 and 2 were core proteins among the DPT-associated proteins.…”