Exp Mol Med.  2010 Nov;42(11):777-786. 10.3858/emm.2010.42.11.079.

Endoplasmic reticulum stress (ER-stress) by 2-deoxy-D-glucose (2DG) reduces cyclooxygenase-2 (COX-2) expression and N-glycosylation and induces a loss of COX-2 activity via a Src kinase-dependent pathway in rabbit articular chondrocytes

• Yu SM
• Kim SJ

• KMID: 994401
• DOI: http://doi.org/10.3858/emm.2010.42.11.079

Abstract

Endoplasmic reticulum (ER) stress regulates a wide range of cellular responses including apoptosis, proliferation, inflammation, and differentiation in mammalian cells. In this study, we observed the role of 2-deoxy-D-glucose (2DG) on inflammation of chondrocytes. 2DG is well known as an inducer of ER stress, via inhibition of glycolysis and glycosylation. Treatment of 2DG in chondrocytes considerably induced ER stress in a dose- and time-dependent manner, which was demonstrated by a reduction of glucose regulated protein of 94 kDa (grp94), an ER stress-inducible protein, as determined by a Western blot analysis. In addition, induction of ER stress by 2DG led to the expression of COX-2 protein with an apparent molecular mass of 66-70kDa as compared with the normally expressed 72-74 kDa protein. The suppression of ER stress with salubrinal (Salub), a selective inhibitor of eif2-alpha dephosphorylation, successfully prevented grp94 induction and efficiently recovered 2DG-modified COX-2 molecular mass and COX-2 activity might be associated with COX-2 N-glycosylation. Also, treatment of 2DG increased phosphorylation of Src in chondrocytes. The inhibition of the Src signaling pathway with PP2 (Src tyrosine kinase inhibitor) suppressed grp94 expression and restored COX-2 expression, N-glycosylation, and PGE2 production, as determined by a Western blot analysis and PGE2 assay. Taken together, our results indicate that the ER stress induced by 2DG results in a decrease of the transcription level, the molecular mass, and the activity of COX-2 in rabbit articular chondrocytes via a Src kinase-dependent pathway.

Keyword

cartilage, articular; chondrocytes; cyclooxygenase 2; deoxyglucose; proto-oncogene proteins pp60(c-src)

MeSH Terms

Animals
Cartilage, Articular/pathology
Cells, Cultured
Chondrocytes/drug effects/immunology/*metabolism/pathology
Cyclooxygenase 2/genetics/*metabolism
Deoxyglucose/*pharmacology
Down-Regulation
Endoplasmic Reticulum/drug effects/*metabolism/pathology
Glycosylation/drug effects
Inflammation
Rabbits
Signal Transduction/drug effects
Stress, Physiological/drug effects/immunology
src-Family Kinases/*metabolism