Reactive oxygen species-dependent nitric oxide production in reciprocal interactions of glioma and microglial cells.

Abstract

Conditioned mediums (CMs) from glioma cells U87, GBM-8401, and C6 significantly induced iNOS protein and NO production by microglial cells BV-2 but without altering the cell viability or cell-cycle progression of BV2 microglia. Significant increases in intracellular peroxide by U87-CM and C6-CM were detected by a DCHF-DA assay, and vitamin (Vit) C and N-acetyl cysteine (NAC)-reduced intracellular peroxide levels elicited by CMs lead to inhibition of iNOS/NO production The extracellular signal-regulated kinase (ERK) inhibitor, U0126, and c-Jun N-terminal kinase (JNK) inhibitor, SP600125, suppressed U87-CM- and C6-CM-induced iNOS/NO production by respectively blocking phosphorylated ERK (pERK) and JNK (pJNK) protein expressions stimulated by U87-CM and C6-CM. Increased migration of U87 and C6 glioma cells by a co-culture with BV-2 microglial cells or adding the nitric oxide (NO) donor, sodium nitroprusside (SNP) was observed, and that was blocked by adding an NO synthase (NOS) inhibitor, N-nitro L-arginine methyl ester (NAME). Contributions of ROS, pERK, and pJNK to the migration of glioma cells was further demonstrated in a transwell coculture system of U87 and C6 gliomas with BV-2 microglial cells. Furthermore, expressions of tumor necrosis factor (TNF)-α and monocyte chemoattractant protein (MCP)-1 messenger (m)RNA in U87 and C6 cells were detected by an RT-PCR, and TNF-α and MCP-1 induced iNOS protein expression in time- and concentration-dependent manners. Neutralization of TNF-α or MCP-1 in U87-CM and C6-CM using a TNF-α or MCP-1 antibody inhibited iNOS protein expression, and increased intracellular peroxide by TNF-α or MCP-1 was identified in BV-2 cells. The reciprocal activation of glioma cells and microglia via ROS-dependent iNOS/NO elevation at least partially mediated by TNF-α and MCP-1 is elucidated.

• Authors: Shing-Chuan Shen, Ming-Shun Wu, Hui-Yi Lin, Liang-Yo Yang, Yi-Hsuan Chen, Yen-Chou Chen
• Journal: Journal of cellular physiology (J Cell Physiol) Vol. 229 Issue 12 Pg. 2015-26 (Dec 2014) ISSN: 1097-4652 [Electronic] United States
• PMID: 24777714 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2014 Wiley Periodicals, Inc.

Chemical References

• Anthracenes
• Butadienes
• CCL2 protein, human
• Chemokine CCL2
• Culture Media, Conditioned
• NF-kappa B
• Nitriles
• Reactive Oxygen Species
• Tumor Necrosis Factor-alpha
• U 0126
• pyrazolanthrone
• Nitric Oxide
• NOS2 protein, human
• Nitric Oxide Synthase Type II
• JNK Mitogen-Activated Protein Kinases

Topics

• Anthracenes (administration & dosage)
• Butadienes (administration & dosage)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Chemokine CCL2 (metabolism)
• Coculture Techniques
• Culture Media, Conditioned
• Gene Expression Regulation, Neoplastic (drug effects)
• Glioma (metabolism, pathology)
• Humans
• JNK Mitogen-Activated Protein Kinases (metabolism)
• Microglia (metabolism, pathology)
• NF-kappa B (biosynthesis, metabolism)
• Nitric Oxide (metabolism)
• Nitric Oxide Synthase Type II (metabolism)
• Nitriles (administration & dosage)
• Reactive Oxygen Species (metabolism)
• Tumor Necrosis Factor-alpha (biosynthesis)