N-Acetyl-Glucosamine Sensitizes Non-Small Cell Lung Cancer Cells to TRAIL-Induced Apoptosis by Activating Death Receptor 5.

Abstract

BACKGROUND/AIMS:

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potential anti-cancer agent due to its selective toxicity. However, many human non-small cell lung cancer (NSCLC) cells are partially resistant to TRAIL, thereby limiting its clinical application. Therefore, there is a need for the development of novel adjuvant therapeutic agents to be used in combination with TRAIL.

METHODS:

In this study, the effect of N-acetyl-glucosamine (GlcNAc), a type of monosaccharide derived from chitosan, combined with TRAIL was evaluated in vitro and in vivo. Thirty NSCLC clinical samples were used to detect the expression of death receptor (DR) 4 and 5. After GlcNAc and TRAIL co-treatment, DR expression was determined by real-time PCR and western blotting. Cycloheximide was used to detect the protein half-life to further understand the correlation between GlcNAc and the metabolic rate of DR. Non-reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to detect receptor clustering, and the localization of DR was visualized by immunofluorescence under a confocal microscope. Furthermore, a co-immunoprecipitation assay was performed to analyze the formation of death-inducing signaling complex (DISC). O-linked glycan expression levels were evaluated following DR5 overexpression and RNA interference mediated knockdown.

RESULTS:

We found that the clinical samples expressed higher levels of DR5 than DR4, and GlcNAc co-treatment improved the effect of TRAIL-induced apoptosis by activating DR5 accumulation and clustering, which in turn recruited the apoptosis-initiating protease caspase-8 to form DISC, and initiated apoptosis. Furthermore, GlcNAc promoted DR5 clustering by improving its O-glycosylation.

CONCLUSION:

These results uncovered the molecular mechanism by which GlcNAc sensitizes cancer cells to TRAIL-induced apoptosis, thereby highlighting a novel effective agent for TRAIL-mediated NSCLC-targeted therapy.

• Authors: Ye Liang, Wenhua Xu, Shihai Liu, Jingwei Chi, Jisheng Zhang, Aihua Sui, Liping Wang, Zhijuan Liang, Dan Li, Yuanbin Chen, Haitao Niu
• Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (Cell Physiol Biochem) Vol. 45 Issue 5 Pg. 2054-2070 ( 2018) ISSN: 1421-9778 [Electronic] Germany
• PMID: 29533936 (Publication Type: Journal Article)
• Copyright: © 2018 The Author(s). Published by S. Karger AG, Basel.

Chemical References

• RNA, Small Interfering
• Receptors, TNF-Related Apoptosis-Inducing Ligand
• TNF-Related Apoptosis-Inducing Ligand
• Poly(ADP-ribose) Polymerases
• Caspase 8
• Acetylglucosamine

Topics

• A549 Cells
• Acetylglucosamine (pharmacology, therapeutic use)
• Animals
• Apoptosis (drug effects)
• Carcinoma, Non-Small-Cell Lung (drug therapy, metabolism, pathology)
• Caspase 8 (metabolism)
• Cell Line, Tumor
• Glycosylation (drug effects)
• Humans
• Immunoprecipitation
• Lung Neoplasms (drug therapy, metabolism, pathology)
• Mice
• Mice, Nude
• Microscopy, Confocal
• Poly(ADP-ribose) Polymerases (metabolism)
• RNA Interference
• RNA, Small Interfering (metabolism)
• Real-Time Polymerase Chain Reaction
• Receptors, TNF-Related Apoptosis-Inducing Ligand (antagonists & inhibitors, genetics, metabolism)
• TNF-Related Apoptosis-Inducing Ligand (therapeutic use, toxicity)
• Transplantation, Heterologous
• Up-Regulation (drug effects)